International Journal of Molecular Sciences

17 pages, 2741 KiB

Open AccessArticle

Quantum Dots-Based Immunofluorescent Imaging of Stromal Fibroblasts Caveolin-1 and Light Chain 3B Expression and Identification of Their Clinical Significance in Human Gastric Cancer

by Yuyu He^1,†, Xianda Zhao^1,†, Jun Gao², Lifang Fan¹, Guifang Yang³, William Chi-shing Cho^4,*

and Honglei Chen^1,5,*

¹ Department of Pathology, School of Basic Medical Science, Wuhan University, Wuhan 430071, China

² Department of Molecular Pathology, Wuhan Nano Tumor Diagnosis Engineering Research Center, Wuhan 430075, China

³ Department of Pathology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China

⁴ Department of Clinical Oncology, Queen Elizabeth Hospital, 30 Gascoigne Road, Kowloon, Hong Kong

⁵ Department of Biochemistry, Rush University Medical Center, 1735 West Harrison Street, Chicago, IL 60612, USA

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 13764-13780; https://doi.org/10.3390/ijms131113764 - 24 Oct 2012

Cited by 33 | Viewed by 7931

Abstract

Caveolin-1 (Cav-1) expression deficiency and autophagy in tumor stromal fibroblasts (hereafter fibroblasts) are involved in tumor proliferation and progression, particularly in breast and prostate cancer. The aim of this study was to detect the expression of fibroblastic Cav-1 and LC3B, markers of autophagy, [...] Read more.

Caveolin-1 (Cav-1) expression deficiency and autophagy in tumor stromal fibroblasts (hereafter fibroblasts) are involved in tumor proliferation and progression, particularly in breast and prostate cancer. The aim of this study was to detect the expression of fibroblastic Cav-1 and LC3B, markers of autophagy, in gastric cancer (GC) and to analyze their clinical significances. Furthermore, because Epstein-Barr virus (EBV)-associated GC (EBVaGC) is a unique subtype of GC; we compared the differential expression of fibroblastic Cav-1 and LC3B in EBVaGC and non-EBVaGC. Quantum dots (QDs)-based immunofluorescence histochemistry was used to examine the expression of fibroblastic Cav-1 and LC3B in 118 cases of GC with adequate stroma. QDs-based double immunofluorescence labeling was performed to detect the coexpression of Cav-1 and LC3B proteins. EBV-encoded small RNA was detected by QDs-based fluorescence in situ hybridization to identify EBVaGC. Multivariate analysis indicated that low fibroblastic Cav-1 level was an independent prognosticator (p = 0.029) that predicted poorer survival of GC patients. Positive fibroblastic LC3B was correlated with lower invasion (p = 0.032) and was positively associated with Cav-1 expression (r = 0.432, p < 0.001). EBV infection did not affect fibroblastic Cav-1 and LC3B expression. In conclusion, positive fibroblastic LC3B correlates with lower invasion, and low expression of fibroblastic Cav-1 is a novel predictor of poor GC prognosis. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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23 pages, 3098 KiB

Open AccessArticle

Impairment of Coronary Arteriolar Endothelium-Dependent Dilation after Multi-Walled Carbon Nanotube Inhalation: A Time-Course Study

by Phoebe A. Stapleton^1,2, Valerie C. Minarchick^1,2, Amy M. Cumpston³, Walter McKinney³, Bean T. Chen³, Tina M. Sager³, David G. Frazer^2,3, Robert R. Mercer³, James Scabilloni³, Michael E. Andrew⁴, Vincent Castranova³ and Timothy R. Nurkiewicz^1,2,3,5,*

¹ Center for Cardiovascular and Respiratory Sciences, West Virginia University School of Medicine, Morgantown, WV 26506, USA

² Department of Physiology and Pharmacology, West Virginia University School of Medicine, Morgantown, WV 26506, USA

³ Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV 26505, USA

⁴ Biostatistics and Epidemiology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV 26505, USA

⁵ Department of Neurobiology and Anatomy, West Virginia University School of Medicine, Morgantown, WV 26505, USA

Int. J. Mol. Sci. 2012, 13(11), 13781-13803; https://doi.org/10.3390/ijms131113781 - 24 Oct 2012

Cited by 97 | Viewed by 8528

Abstract

Engineered nanomaterials have been developed for widespread applications due to many highly unique and desirable characteristics. The purpose of this study was to assess pulmonary inflammation and subepicardial arteriolar reactivity in response to multi-walled carbon nanotube (MWCNT) inhalation and evaluate the time course [...] Read more.

Engineered nanomaterials have been developed for widespread applications due to many highly unique and desirable characteristics. The purpose of this study was to assess pulmonary inflammation and subepicardial arteriolar reactivity in response to multi-walled carbon nanotube (MWCNT) inhalation and evaluate the time course of vascular alterations. Rats were exposed to MWCNT aerosols producing pulmonary deposition. Pulmonary inflammation via bronchoalveolar lavage and MWCNT translocation from the lungs to systemic organs was evident 24 h post-inhalation. Coronary arterioles were evaluated 24–168 h post-exposure to determine microvascular response to changes in transmural pressure, endothelium-dependent and -independent reactivity. Myogenic responsiveness, vascular smooth muscle reactivity to nitric oxide, and α-adrenergic responses all remained intact. However, a severe impact on endothelium-dependent dilation was observed within 24 h after MWCNT inhalation, a condition which improved, but did not fully return to control after 168 h. In conclusion, results indicate that MWCNT inhalation not only leads to pulmonary inflammation and cytotoxicity at low lung burdens, but also a low level of particle translocation to systemic organs. MWCNT inhalation also leads to impairments of endothelium-dependent dilation in the coronary microcirculation within 24 h, a condition which does not fully dissipate within 168 h. The innovations within the field of nanotechnology, while exciting and novel, can only reach their full potential if toxicity is first properly assessed. Full article

(This article belongs to the Special Issue Bioactive Nanoparticles 2012)

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12 pages, 260 KiB

Open AccessArticle

Asymmetric Dimethylarginine Plasma Levels and Endothelial Function in Newly Diagnosed Type 2 Diabetic Patients

by Angela Sciacqua, Nadia Grillo, Michele Quero, Giorgio Sesti

and Francesco Perticone^*

Department of Medical and Surgical Sciences, University Magna Græcia of Catanzaro, Campus Germaneto, viale Europa, 88100, Italy

Int. J. Mol. Sci. 2012, 13(11), 13804-13815; https://doi.org/10.3390/ijms131113804 - 24 Oct 2012

Cited by 29 | Viewed by 5485

Abstract

It is now well established that major risk factors for cardiovascular diseases (CVD) impact upon endothelial function by decreasing nitric oxide (NO) bioavailability. Asymmetric dimethylarginine (ADMA), an endogenous analog of l-arginine, is able to inhibit the activity of endothelial-NO synthase, promoting endothelial dysfunction. [...] Read more.

It is now well established that major risk factors for cardiovascular diseases (CVD) impact upon endothelial function by decreasing nitric oxide (NO) bioavailability. Asymmetric dimethylarginine (ADMA), an endogenous analog of l-arginine, is able to inhibit the activity of endothelial-NO synthase, promoting endothelial dysfunction. Type 2 diabetes (T2D) is characterized by a reduced endothelium-dependent vasodilation and increased ADMA levels and ADMA is strongly associated with micro- and macrovascular diabetic complications. However, there are not a lot of data about the role of ADMA on endothelial function in newly diagnosed T2D patients without cardiovascular (CV) complications. For this aim, we have enrolled forty-five newly diagnosed T2D patients, evaluated by a oral glucose tolerance test, and thirty normal subjects. Endothelium-dependent and -independent vasodilatation was investigated by intra-arterial infusion of increasing doses of acetylcholine (ACh) and sodium nitroprusside. ADMA was measured by high-performance liquid chromatography and insulin resistance (IR) by HOMA. Newly diagnosed T2D patients showed higher ADMA and l-arginine mean values in comparison with normal subjects and a significantly reduced ACh-stimulated forearm blood flow (FBF). In T2D patients FBF was significantly and inversely correlated with ADMA (r = −0.524, p < 0.0001) and in a multivariate regression analysis, ADMA resulted the stronger predictor of FBF, explaining the 27.5% of variability (p < 0.0001). In conclusion, ADMA was strongly related to endothelial dysfunction also in patients with newly diagnosed T2D, without clinically manifest vascular complications. This field is of great interest for understanding the mechanisms underlying the pathogenesis of diabetic disease and its CV complications. Full article

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14 pages, 795 KiB

Open AccessArticle

Phorbol Esters Isolated from Jatropha Meal Induced Apoptosis-Mediated Inhibition in Proliferation of Chang and Vero Cell Lines

by Ehsan Oskoueian^1,2, Norhani Abdullah^3,4,* and Syahida Ahmad³

¹ Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

² Agriculture Biotechnology Research Institute of Iran (ABRII)-East and North-East Branch, P.O.B. 91735/844, Mashhad, Iran

³ Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

⁴ Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 13816-13829; https://doi.org/10.3390/ijms131113816 - 24 Oct 2012

Cited by 16 | Viewed by 7834

Abstract

The direct feeding of Jatropha meal containing phorbol esters (PEs) indicated mild to severe toxicity symptoms in various organs of different animals. However, limited information is available on cellular and molecular mechanism of toxicity caused by PEs present in Jatropha meal. Thus, the [...] Read more.

The direct feeding of Jatropha meal containing phorbol esters (PEs) indicated mild to severe toxicity symptoms in various organs of different animals. However, limited information is available on cellular and molecular mechanism of toxicity caused by PEs present in Jatropha meal. Thus, the present study was conducted to determine the cytotoxic and mode of action of PEs isolated from Jatropha meal using human hepatocyte (Chang) and African green monkey kidney (Vero) cell lines. The results showed that isolated PEs inhibited cell proliferation in a dose-dependent manner in both cell lines with the CC₅₀ of 125.9 and 110.3 μg/mL, respectively. These values were compatible to that of phorbol 12-myristate 13-acetate (PMA) values as positive control i.e., 124.5 and 106.3 μg/mL respectively. Microscopic examination, flow cytometry and DNA fragmentation results confirmed cell death due to apoptosis upon treatment with PEs and PMA at CC₅₀ concentration for 24 h in both cell lines. The Western blot analysis revealed the overexpression of PKC-δ and activation of caspase-3 proteins which could be involved in the mechanism of action of PEs and PMA. Consequently, the PEs isolated form Jatropha meal caused toxicity and induced apoptosis-mediated proliferation inhibition toward Chang and Vero cell lines involving over-expression of PKC-δ and caspase-3 as their mode of actions. Full article

(This article belongs to the Section Biochemistry)

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37 pages, 275 KiB

Open AccessArticle

Oxidant Stress and Signal Transduction in the Nervous System with the PI 3-K, Akt, and mTOR Cascade

by Kenneth Maiese^1,2,3,*, Zhao Zhong Chong^1,3, Shaohui Wang^1,3 and Yan Chen Shang^1,3

¹ Laboratory of Cellular and Molecular Signaling, Newark, NJ 07101, USA

² Cancer Institute of New Jersey, New Brunswick, NJ 08903, USA

³ New Jersey Health Sciences University, 205 South Orange Avenue, Newark, NJ 07101, USA

Int. J. Mol. Sci. 2012, 13(11), 13830-13866; https://doi.org/10.3390/ijms131113830 - 26 Oct 2012

Cited by 96 | Viewed by 18402

Abstract

Oxidative stress impacts multiple systems of the body and can lead to some of the most devastating consequences in the nervous system especially during aging. Both acute and chronic neurodegenerative disorders such as diabetes mellitus, cerebral ischemia, trauma, Alzheimer’s disease, Parkinson’s disease, Huntington’s [...] Read more.

Oxidative stress impacts multiple systems of the body and can lead to some of the most devastating consequences in the nervous system especially during aging. Both acute and chronic neurodegenerative disorders such as diabetes mellitus, cerebral ischemia, trauma, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and tuberous sclerosis through programmed cell death pathways of apoptosis and autophagy can be the result of oxidant stress. Novel therapeutic avenues that focus upon the phosphoinositide 3-kinase (PI 3-K), Akt (protein kinase B), and the mammalian target of rapamycin (mTOR) cascade and related pathways offer exciting prospects to address the onset and potential reversal of neurodegenerative disorders. Effective clinical translation of these pathways into robust therapeutic strategies requires intimate knowledge of the complexity of these pathways and the ability of this cascade to influence biological outcome that can vary among disorders of the nervous system. Full article

(This article belongs to the Special Issue Oxidative Stress and Ageing)

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Graphical abstract

14 pages, 754 KiB

Open AccessArticle

Enhancement of Commercial Antifungal Agents by Kojic Acid

by Jong H. Kim^1,*

, Perng-Kuang Chang², Kathleen L. Chan¹, Natália C. G. Faria³, Noreen Mahoney¹, Young K. Kim^1,4, Maria De L. Martins³ and Bruce C. Campbell¹

¹ Plant Mycotoxin Research Unit, Western Regional Research Center, USDA-ARS, 800 Buchanan St., Albany, CA 94710, USA

² Food and Feed Safety Research Unit, Southern Regional Research Center, USDA-ARS, 1100 Robert E. Lee Blvd., New Orleans, LA 70124, USA

³ Instituto de Higiene e Medicina Tropical/CREM, Universidade Nova de Lisboa, Portugal

⁴ Department of Forest Products and Biotechnology, College of Forest Sciences, Kookmin University, Seoul 136-702, Korea

Int. J. Mol. Sci. 2012, 13(11), 13867-13880; https://doi.org/10.3390/ijms131113867 - 26 Oct 2012

Cited by 32 | Viewed by 9417

Abstract

Natural compounds that pose no significant medical or environmental side effects are potential sources of antifungal agents, either in their nascent form or as structural backbones for more effective derivatives. Kojic acid (KA) is one such compound. It is a natural by-product of [...] Read more.

Natural compounds that pose no significant medical or environmental side effects are potential sources of antifungal agents, either in their nascent form or as structural backbones for more effective derivatives. Kojic acid (KA) is one such compound. It is a natural by-product of fungal fermentation commonly employed by food and cosmetic industries. We show that KA greatly lowers minimum inhibitory (MIC) or fungicidal (MFC) concentrations of commercial medicinal and agricultural antifungal agents, amphotericin B (AMB) and strobilurin, respectively, against pathogenic yeasts and filamentous fungi. Assays using two mitogen-activated protein kinase (MAPK) mutants, i.e., sakA∆, mpkC∆, of Aspergillus fumigatus, an agent for human invasive aspergillosis, with hydrogen peroxide (H₂O₂) or AMB indicate such chemosensitizing activity of KA is most conceivably through disruption of fungal antioxidation systems. KA could be developed as a chemosensitizer to enhance efficacy of certain conventional antifungal drugs or fungicides. Full article

(This article belongs to the Section Biochemistry)

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13 pages, 1866 KiB

Open AccessArticle

The Role of Forkhead Box Q1 Transcription Factor in Ovarian Epithelial Carcinomas

by Min Gao^1,2, Ie-Ming Shih² and Tian-Li Wang^2,*

¹ Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, 107 W. Wenhua Road, Jinan 250012, China

² Departments of Pathology, Gynecology/Obstetrics, and Oncology, Program in Pathobiology, Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA

Int. J. Mol. Sci. 2012, 13(11), 13881-13893; https://doi.org/10.3390/ijms131113881 - 26 Oct 2012

Cited by 33 | Viewed by 8062

Abstract

The role of the forkhead box Q1 (FOXQ1) transcription factor in cancer pathogenesis has recently emerged. Overexpression of FOXQ1 has been found in a variety of human cancers, and its upregulation has been associated with poor prognosis in colorectal, breast, and non-small cell [...] Read more.

The role of the forkhead box Q1 (FOXQ1) transcription factor in cancer pathogenesis has recently emerged. Overexpression of FOXQ1 has been found in a variety of human cancers, and its upregulation has been associated with poor prognosis in colorectal, breast, and non-small cell lung carcinomas. However, the molecular mechanism underlying how FOXQ1 contributes to ovarian epithelial carcinomas remains unclear. To this end, we analyzed gene expression levels in ovarian cancer tissues and cell lines and demonstrated a higher expression level of FOXQ1 in epithelial ovarian cancer cells than that in normal epithelial cells. We then used a human ovarian cancer cell line, SKOV3, which expressed a higher level of FOXQ1, as a cell model to investigate the biological effects of FOXQ1 by using RNA interference. Silencing of FOXQ1 expression using a shRNA knockdown approach affected the expression of several cell cycle regulators, leading to suppressed cell proliferation, reduced cell motility/invasion, and upregulation of epithelial cell markers and the downregulation of mesenchymal cell markers. Taken together, these results suggest that FOXQ1 expression is essential to maintain cell proliferation, motility/invasion, and epithelial-mesenchymal transition phenotypes in ovarian cancer cells. Full article

(This article belongs to the Special Issue Genes and Pathways in the Pathogenesis of Ovarian Cancer)

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17 pages, 1251 KiB

Open AccessArticle

Profiling the Proteome of Exhaled Breath Condensate in Healthy Smokers and COPD Patients by LC-MS/MS

by Marco Fumagalli¹, Fabio Ferrari², Maurizio Luisetti³, Jan Stolk⁴, Pieter S. Hiemstra⁴, Daniela Capuano³, Simona Viglio⁵, Laura Fregonese⁴, Isa Cerveri³, Federica Corana⁶, Carmine Tinelli⁷ and Paolo Iadarola^1,*

¹ Department of Biology and Biotechnology, University of Pavia, Via Taramelli 3/B, 27100 Pavia, Italy

² Lab Analysis Inc., Casanova Lonati, 27041 Pavia, Italy

³ Department of Molecular Medicine, Division of Pneumology, University of Pavia & IRCCS Policlinico San Matteo, Via Taramelli 5, 27100 Pavia, Italy

⁴ Department of Pulmonology, Leiden University Medical Center, 2333 Leiden, The Netherlands

⁵ Department of Molecular Medicine, Division of Biochemistry, University of Pavia, Via Taramelli 3/B, 27100, Pavia, Italy

⁶ Centro Grandi Strumenti, University of Pavia, Via Bassi 6, 27100 Pavia, Italy

⁷ Biometric Unit, IRCCS San Matteo Hospital Foundation, 27100 Pavia, Italy

Int. J. Mol. Sci. 2012, 13(11), 13894-13910; https://doi.org/10.3390/ijms131113894 - 29 Oct 2012

Cited by 55 | Viewed by 9404

Abstract

Three pools of exhaled breath condensate (EBC) from non-smokers plus healthy smokers (NS + HS, n = 45); chronic obstructive pulmonary disease (COPD) without emphysema (COPD, n = 15) and subjects with pulmonary emphysema associated with α₁-antitrypsin deficiency (AATD, n = [...] Read more.

Three pools of exhaled breath condensate (EBC) from non-smokers plus healthy smokers (NS + HS, n = 45); chronic obstructive pulmonary disease (COPD) without emphysema (COPD, n = 15) and subjects with pulmonary emphysema associated with α₁-antitrypsin deficiency (AATD, n = 23) were used for an exploratory proteomic study aimed at generating fingerprints of these groups that can be used in future pathophysiological and perhaps even clinical research. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was the platform applied for this hypothesis-free investigation. Analysis of pooled specimens resulted in the production of a “fingerprint” made of 44 proteins for NS/HS; 17 for COPD and 15 for the group of AATD subjects. Several inflammatory cytokines (IL-1α, IL-1β, IL-2; IL-12, α and β subunits, IL-15, interferon α and γ, tumor necrosis factor α); Type I and II cytokeratins; two SP-A isoforms; Calgranulin A and B and α1-antitrypsin were detected and validated through the use of surface enhanced laser-desorption ionization mass spectrometry (SELDI-MS) and/or by Western blot (WB) analysis. These results are the prelude of quantitative studies aimed at identifying which of these proteins hold promise as identifiers of differences that could distinguish healthy subjects from patients. Full article

(This article belongs to the Special Issue Advances in Proteomic Research)

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Graphical abstract

15 pages, 395 KiB

Open AccessArticle

Injurious Effects of Emodin on Maturation of Mouse Oocytes, Fertilization and Fetal Development via Apoptosis

by Mei-Hui Chang, Shao-Chung Chang and Wen-Hsiung Chan^*

Department of Bioscience Technology and Center for Nanotechnology, Chung Yuan Christian University, Chung Li 32023, Taiwan

Int. J. Mol. Sci. 2012, 13(11), 13911-13925; https://doi.org/10.3390/ijms131113911 - 29 Oct 2012

Cited by 16 | Viewed by 6072

Abstract

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Previous studies have established that emodin induces apoptosis in the inner cell mass and trophectoderm of mouse blastocysts and leads to decreased embryonic development and viability, indicating a role [...] Read more.

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Previous studies have established that emodin induces apoptosis in the inner cell mass and trophectoderm of mouse blastocysts and leads to decreased embryonic development and viability, indicating a role as an injury risk factor for normal embryonic development. However, the mechanisms underlying its hazardous effects have yet to be characterized. In the current study, we further investigated the effects of emodin on oocyte maturation and subsequent pre- and post-implantation development, both in vitro and in vivo. Notably, emodin induced a significant reduction in the rates of oocyte maturation, fertilization, and in vitro embryonic development. Treatment of oocytes with emodin during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased fetal weight. Experiments using an in vivo mouse model disclosed that consumption of drinking water containing 20–40 μM emodin led to decreased oocyte maturation and in vitro fertilization, as well as early embryonic developmental injury. Notably, pretreatment with a caspase-3-specific inhibitor effectively prevented emodin-triggered injury effects, suggesting that impairment of embryo development occurs via a caspase-dependent apoptotic process. Full article

(This article belongs to the Collection Programmed Cell Death and Apoptosis)

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23 pages, 877 KiB

Open AccessReview

The Proteomics Big Challenge for Biomarkers and New Drug-Targets Discovery

by Rocco Savino, Sergio Paduano, Mariaimmacolata Preianò and Rosa Terracciano^*

Department of Health Sciences, Laboratory of Mass Spectrometry and Proteomics, University “Magna Græcia”, Catanzaro, University Campus, Europa Avenue, 88100 Catanzaro, Italy

Int. J. Mol. Sci. 2012, 13(11), 13926-13948; https://doi.org/10.3390/ijms131113926 - 29 Oct 2012

Cited by 64 | Viewed by 12667

Abstract

In the modern process of drug discovery, clinical, functional and chemical proteomics can converge and integrate synergies. Functional proteomics explores and elucidates the components of pathways and their interactions which, when deregulated, lead to a disease condition. This knowledge allows the design of [...] Read more.

In the modern process of drug discovery, clinical, functional and chemical proteomics can converge and integrate synergies. Functional proteomics explores and elucidates the components of pathways and their interactions which, when deregulated, lead to a disease condition. This knowledge allows the design of strategies to target multiple pathways with combinations of pathway-specific drugs, which might increase chances of success and reduce the occurrence of drug resistance. Chemical proteomics, by analyzing the drug interactome, strongly contributes to accelerate the process of new druggable targets discovery. In the research area of clinical proteomics, proteome and peptidome mass spectrometry-profiling of human bodily fluid (plasma, serum, urine and so on), as well as of tissue and of cells, represents a promising tool for novel biomarker and eventually new druggable targets discovery. In the present review we provide a survey of current strategies of functional, chemical and clinical proteomics. Major issues will be presented for proteomic technologies used for the discovery of biomarkers for early disease diagnosis and identification of new drug targets. Full article

(This article belongs to the Special Issue Advances in Proteomic Research)

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17 pages, 769 KiB

Open AccessReview

Microenvironments and Cellular Characteristics in the Micro Tumor Cords of Malignant Solid Tumors

by Chan Joo Yeom¹, Yoko Goto^1,2, Yuxi Zhu^1,2,3, Masahiro Hiraoka² and Hiroshi Harada^1,*

¹ Group of Radiation and Tumor Biology, Career-Path Promotion Unit for Young Life Scientists, Kyoto University, Yoshida Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan

² Department of Radiation Oncology and Image-applied Therapy, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan

³ Department of Oncology, The First Affiliated Hospital of Chongqing Medical University, No.1 Friendship Road, Yuanjiagang, Yuzhong District, Chongqing 400016, China

Int. J. Mol. Sci. 2012, 13(11), 13949-13965; https://doi.org/10.3390/ijms131113949 - 29 Oct 2012

Cited by 55 | Viewed by 9245

Abstract

Because of the accelerated proliferation of cancer cells and the limited distance that molecular oxygen can diffuse from functional tumor blood vessels, there appears to be a unique histology in malignant solid tumors, conglomerates of micro tumor cords. A functional blood vessel exists [...] Read more.

Because of the accelerated proliferation of cancer cells and the limited distance that molecular oxygen can diffuse from functional tumor blood vessels, there appears to be a unique histology in malignant solid tumors, conglomerates of micro tumor cords. A functional blood vessel exists at the center of each tumor cord and is sequentially surrounded by well-oxygenated, oxygen-insufficient, and oxygen-depleted cancer cells in the shape of baumkuchen (layered). Cancer cells, by inducing the expression of various genes, adapt to the highly heterogeneous microenvironments in each layer. Accumulated evidence has suggested that not only tumor microenvironments but also cellular adaptive responses to them, influence the radioresistance of cancer cells. However, precisely how these factors affect one another and eventually influence the therapeutic effect of radiation therapy remains to be elucidated. Here, based on recent basic and clinical cancer research, we deduced extrinsic (oxygen concentration, glucose concentration, pH etc.) and intrinsic (transcriptional activity of hypoxia-inducible factor 1, metabolic pathways, cell cycle status, proliferative activity etc.) parameters in each layer of a tumor cord. In addition, we reviewed the latest information about the molecular mechanism linking these factors with both tumor radioresistance and tumor recurrence after radiation therapy. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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19 pages, 1185 KiB

Open AccessArticle

Novel High-Viscosity Polyacrylamidated Chitosan for Neural Tissue Engineering: Fabrication of Anisotropic Neurodurable Scaffold via Molecular Disposition of Persulfate-Mediated Polymer Slicing and Complexation

by Pradeep Kumar¹

, Yahya E. Choonara¹

, Lisa C. du Toit¹, Girish Modi², Dinesh Naidoo³ and Viness Pillay^1,*

¹ Department of Pharmacy and Pharmacology, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, 2193, Johannesburg, South Africa

² Department of Neurology, Division of Neurosciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, 2193, Johannesburg, South Africa

³ Department of Neurosurgery, Division of Neurosciences, Faculty of Health Sciences, University of the Witwatersrand, 7 York Road, Parktown, 2193, Johannesburg, South Africa

Int. J. Mol. Sci. 2012, 13(11), 13966-13984; https://doi.org/10.3390/ijms131113966 - 29 Oct 2012

Cited by 59 | Viewed by 13774

Abstract

Macroporous polyacrylamide-grafted-chitosan scaffolds for neural tissue engineering were fabricated with varied synthetic and viscosity profiles. A novel approach and mechanism was utilized for polyacrylamide grafting onto chitosan using potassium persulfate (KPS) mediated degradation of both polymers under a thermally controlled environment. Commercially available [...] Read more.

Macroporous polyacrylamide-grafted-chitosan scaffolds for neural tissue engineering were fabricated with varied synthetic and viscosity profiles. A novel approach and mechanism was utilized for polyacrylamide grafting onto chitosan using potassium persulfate (KPS) mediated degradation of both polymers under a thermally controlled environment. Commercially available high molecular mass polyacrylamide was used instead of the acrylamide monomer for graft copolymerization. This grafting strategy yielded an enhanced grafting efficiency (GE = 92%), grafting ratio (GR = 263%), intrinsic viscosity (IV = 5.231 dL/g) and viscometric average molecular mass (MW = 1.63 × 10⁶ Da) compared with known acrylamide that has a GE = 83%, GR = 178%, IV = 3.901 dL/g and MW = 1.22 × 10⁶ Da. Image processing analysis of SEM images of the newly grafted neurodurable scaffold was undertaken based on the polymer-pore threshold. Attenuated Total Reflectance-FTIR spectral analyses in conjugation with DSC were used for the characterization and comparison of the newly grafted copolymers. Static Lattice Atomistic Simulations were employed to investigate and elucidate the copolymeric assembly and reaction mechanism by exploring the spatial disposition of chitosan and polyacrylamide with respect to the reactional profile of potassium persulfate. Interestingly, potassium persulfate, a peroxide, was found to play a dual role initially degrading the polymers—“polymer slicing”—thereby initiating the formation of free radicals and subsequently leading to synthesis of the high molecular mass polyacrylamide-grafted-chitosan (PAAm-g-CHT)—“polymer complexation”. Furthermore, the applicability of the uniquely grafted scaffold for neural tissue engineering was evaluated via PC12 neuronal cell seeding. The novel PAAm-g-CHT exhibited superior neurocompatibility in terms of cell infiltration owing to the anisotropic porous architecture, high molecular mass mediated robustness, superior hydrophilicity as well as surface charge due to the acrylic chains. Additionally, these results suggested that the porous PAAm-g-CHT scaffold may act as a potential neural cell carrier. Full article

(This article belongs to the Section Materials Science)

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17 pages, 608 KiB

Open AccessArticle

Superoxide Dismutase as a Novel Macromolecular Nitric Oxide Carrier: Preparation and Characterization

by Ssu-Han Chen¹, Shih-Jiuan Chiu² and Teh-Min Hu^1,*

¹ School of Pharmacy, National Defense Medical Center, Taipei 11490, Taiwan

² School of Pharmacy, Taipei Medical University, Taipei 11031, Taiwan

Int. J. Mol. Sci. 2012, 13(11), 13985-14001; https://doi.org/10.3390/ijms131113985 - 29 Oct 2012

Cited by 2 | Viewed by 6795

Abstract

Nitric oxide (NO) is an important molecule that exerts multiple functions in biological systems. Because of the short-lived nature of NO, S-nitrosothiols (RSNOs) are believed to act as stable NO carriers. Recently, sulfhydryl (SH) containing macromolecules have been shown to be promising [...] Read more.

Nitric oxide (NO) is an important molecule that exerts multiple functions in biological systems. Because of the short-lived nature of NO, S-nitrosothiols (RSNOs) are believed to act as stable NO carriers. Recently, sulfhydryl (SH) containing macromolecules have been shown to be promising NO carriers. In the present study, we aimed to synthesize and characterize a potential NO carrier based on bovine Cu,Zn-superoxide dismutase (bSOD). To prepare S-nitrosated bSOD, the protein was incubated with S-nitrosoglutathione (GSNO) under varied experimental conditions. The results show that significant S-nitrosation of bSOD occurred only at high temperature (50 °C) for prolonged incubation time (>2 h). S-nitrosation efficiency increased with reaction time and reached a plateau at ~4 h. The maximum amount of NO loaded was determined to be about 0.6 mol SNO/mol protein (~30% loading efficiency). The enzymatic activity of bSOD, however, decreased with reaction time. Our data further indicate that NO functionality can only be measured in the presence of extremely high concentrations of Hg²⁺ or when the protein was denatured by guanidine. Moreover, mildly acidic pH was shown to favor S-nitrosation of bSOD. A model based on unfolding and refolding of bSOD during preparation was proposed to possibly explain our observation. Full article

(This article belongs to the Special Issue Advances in Free Radicals in Biology and Medicine)

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14 pages, 115 KiB

Open AccessReview

Bacterial Exopolysaccharides: Functionality and Prospects

by Uchechukwu U. Nwodo^*, Ezekiel Green and Anthony I. Okoh

Applied and Environmental Microbiology Research Group (AEMREG), Department of Biochemistry and Microbiology, University of Fort Hare, Private Bag X1314, Alice 5700, South Africa

Int. J. Mol. Sci. 2012, 13(11), 14002-14015; https://doi.org/10.3390/ijms131114002 - 30 Oct 2012

Cited by 540 | Viewed by 22155

Abstract

Diverse structural, functional and valuable polysaccharides are synthesized by bacteria of all taxa and secreted into the external environment. These polysaccharides are referred to as exopolysaccharides and they may either be homopolymeric or heteropolymeric in composition and of diverse high molecular weights (10 [...] Read more.

Diverse structural, functional and valuable polysaccharides are synthesized by bacteria of all taxa and secreted into the external environment. These polysaccharides are referred to as exopolysaccharides and they may either be homopolymeric or heteropolymeric in composition and of diverse high molecular weights (10 to 1000 kDa). The material properties of exopolysaccharides have revolutionized the industrial and medical sectors due to their retinue of functional applications and prospects. These applications have been extensive in areas such as pharmacological, nutraceutical, functional food, cosmeceutical, herbicides and insecticides among others, while prospects includes uses as anticoagulant, antithrombotic, immunomodulation, anticancer and as bioflocculants. Due to the extensive applications of bacterial exopolysaccharides, this overview provides basic information on their physiologic and morphologic functions as well as their applications and prospects in the medical and industrial sectors. Full article

10 pages, 183 KiB

Open AccessArticle

Digluconate and Isopropyl Alcohol Biocide Formulation

by Emma Hendry¹, Barbara Conway²

and Tony Worthington^1,*

¹ Microbiology, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK

² Pharmacy and Pharmaceutical Sciences, School of Applied Sciences, University of Huddersfield, Huddersfield HD9 3JA, UK

Int. J. Mol. Sci. 2012, 13(11), 14016-14025; https://doi.org/10.3390/ijms131114016 - 30 Oct 2012

Cited by 13 | Viewed by 8255

Abstract

Effective surface disinfection is a fundamental infection control strategy within healthcare. This study assessed the antimicrobial efficacy of novel biocide formulations comprising 5% and 2% eucalyptus oil (EO) combined with 2% chlorhexidine digluconate (CHG) and 70% isopropyl alcohol (IPA) contained within a wipe. [...] Read more.

Effective surface disinfection is a fundamental infection control strategy within healthcare. This study assessed the antimicrobial efficacy of novel biocide formulations comprising 5% and 2% eucalyptus oil (EO) combined with 2% chlorhexidine digluconate (CHG) and 70% isopropyl alcohol (IPA) contained within a wipe. The efficacy of this novel antimicrobial formulation to remove and eliminate methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Candida albicans from steel surfaces was investigated. Adpression studies of pre-contaminated wipes were also utilised to assess their potential to induce cross-contamination between hard surfaces. Furthermore, the bactericidal nature of the EO-formulation was established in addition to time-kill. The EO-containing formulations demonstrated bactericidal antimicrobial efficacy against all microorganisms and did not induce surface cross-contamination. There was no significant difference (p < 0.05) between the 5% and 2% EO formulations in their ability to remove microorganisms from steel surfaces, however both significantly (p < 0.05) removed more than the control formulations. Microbial biofilms were eliminated within 10 min (p < 0.05) when exposed to the EO formulations. Our novel EO-formulation demonstrated rapid antimicrobial efficacy for potential disinfection and elimination of microbial biofilms from hard surfaces and may therefore be a useful adjunct to current infection control strategies currently employed within healthcare facilities. Full article

(This article belongs to the Special Issue Green Biocides)

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27 pages, 615 KiB

Open AccessReview

Types, Causes, Detection and Repair of DNA Fragmentation in Animal and Human Sperm Cells

by Clara González-Marín¹, Jaime Gosálvez² and Rosa Roy^2,*

¹ Sexing Technologies, 22575 State Highway 6 South, Navasota, TX 77868, USA

² Biology Department, Universidad Autonoma of Madrid, C/ Darwin nº 2. 28049 Madrid, Spain

Int. J. Mol. Sci. 2012, 13(11), 14026-14052; https://doi.org/10.3390/ijms131114026 - 31 Oct 2012

Cited by 269 | Viewed by 21080

Abstract

Concentration, motility and morphology are parameters commonly used to determine the fertilization potential of an ejaculate. These parameters give a general view on the quality of sperm but do not provide information about one of the most important components of the reproductive outcome: [...] Read more.

Concentration, motility and morphology are parameters commonly used to determine the fertilization potential of an ejaculate. These parameters give a general view on the quality of sperm but do not provide information about one of the most important components of the reproductive outcome: DNA. Either single or double DNA strand breaks can set the difference between fertile and infertile males. Sperm DNA fragmentation can be caused by intrinsic factors like abortive apoptosis, deficiencies in recombination, protamine imbalances or oxidative stress. Damage can also occur due to extrinsic factors such as storage temperatures, extenders, handling conditions, time after ejaculation, infections and reaction to medicines or post-testicular oxidative stress, among others. Two singular characteristics differentiate sperm from somatic cells: Protamination and absence of DNA repair. DNA repair in sperm is terminated as transcription and translation stops post-spermiogenesis, so these cells have no mechanism to repair the damage occurred during their transit through the epididymis and post-ejaculation. Oocytes and early embryos have been shown to repair sperm DNA damage, so the effect of sperm DNA fragmentation depends on the combined effects of sperm chromatin damage and the capacity of the oocyte to repair it. In this contribution we review some of these issues. Full article

(This article belongs to the Section Biochemistry)

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20 pages, 1759 KiB

Open AccessArticle

Profound Re-Organization of Cell Surface Proteome in Equine Retinal Pigment Epithelial Cells in Response to In Vitro Culturing

by Christoph M. Szober¹, Stefanie M. Hauck²

, Kerstin N. Euler¹, Kristina J. H. Fröhlich¹, Claudia Alge-Priglinger³, Marius Ueffing^2,4 and Cornelia A. Deeg^1,*

¹ Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig-Maximilians-University Munich, D-80539 Munich, Germany

² Research Unit Protein Science, Helmholtz Center Munich, German Research Center for Environmental Health, D-85764 Neuherberg, Germany

³ Department of Ophthalmology, Ludwig-Maximilians-University, Mathildenstrasse 8, D-80336 Munich, Germany

⁴ Centre of Ophthalmology, Institute for Ophthalmic Research, University of Tübingen, Röntgenweg 11, D-72076 Tübingen, Germany

Int. J. Mol. Sci. 2012, 13(11), 14053-14072; https://doi.org/10.3390/ijms131114053 - 31 Oct 2012

Cited by 8 | Viewed by 7757

Abstract

The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of [...] Read more.

The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses’ vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, analyzed by high resolution mass spectrometry coupled to liquid chromatography (LC MS/MS), and the most interesting candidates were validated by PCR, immunoblotting and immunocytochemistry. A total of 112 proteins were identified, of which 84% were cell surface membrane proteins. Twenty-three of these proteins were concurrently expressed by both cell states, 28 proteins exclusively by native RPE cells. Among the latter were two RPE markers with highly specialized RPE functions: cellular retinaldehyde-binding protein (CRALBP) and retinal pigment epithelium-specific protein 65kDa (RPE65). Furthermore, 61 proteins were only expressed by cultured RPE cells and absent in native cells. As we believe that initiating events, leading to the breakdown of the outer blood-retinal barrier, take place at the cell surface of RPE cells as a particularly exposed barrier structure, this differential characterization of cell surface proteomes of native and cultured equine RPE cells is a prerequisite for future studies. Full article

(This article belongs to the Special Issue Advances in Proteomic Research)

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13 pages, 5623 KiB

Open AccessArticle

Association of Retinoic Acid Receptors with Extracellular Matrix Accumulation in Rats with Renal Interstitial Fibrosis Disease

by Yao-Bin Long, Yuan-Han Qin^*, Tian-Biao Zhou and Feng-Ying Lei

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China

Int. J. Mol. Sci. 2012, 13(11), 14073-14085; https://doi.org/10.3390/ijms131114073 - 31 Oct 2012

Cited by 16 | Viewed by 5182

Abstract

The nuclear retinoic acid receptors (RARs) function as ligand-dependent transcriptional regulators and include three subtypes (RARα, RARβ and RARγ), which control the expression of specific gene subsets subsequent to ligand binding and to strictly controlled phosphorylation processes. Extracellular matrix (ECM) accumulation is the [...] Read more.

The nuclear retinoic acid receptors (RARs) function as ligand-dependent transcriptional regulators and include three subtypes (RARα, RARβ and RARγ), which control the expression of specific gene subsets subsequent to ligand binding and to strictly controlled phosphorylation processes. Extracellular matrix (ECM) accumulation is the most important characteristic of renal interstitial fibrosis (RIF). This study was performed to investigate whether RARs were associated with ECM accumulation in the progression of RIF in rats. Eighty Wistar male rats were divided into a sham operation group (SHO) and a model group subjected to unilateral ureteral obstruction (GU) at random; n = 40, respectively. The RIF disease in GU group was established by left ureteral ligation. The renal tissues were collected at two weeks and four weeks after surgery. Protein expressions of RARα, RARβ, RARγ, transforming growth factor-βl (TGF-β1), collagen-IV (Col-IV) and fibronectin (FN) were detected using immunohistochemical analysis, and mRNA expressions of RARα, RARβ, RARγ and TGF-β1 in renal tissue were detected by real time reverse transcription polymerase chain reaction. RIF index in renal interstitium was also calculated. When compared with those in SHO group, expressions of RARα and RARβ (protein and mRNA) were markedly reduced in the GU group (each p < 0.01). There was no marked difference for the expression of RARγ (protein and mRNA) between the SHO group and the GU group. The expressions of TGF-β1, Col-IV, FN and the RIF index in the GU group were markedly increased when compared with those in the SHO group (each p < 0.01). The protein expression of RARα/RARβ was negatively correlated with protein expression of TGF-β1, Col-IV or FN and the RIF index (all p < 0.01). In conclusion, the low expression of RARα/RARβ is associated with ECM accumulation in the progression of RIF in rats, suggesting that RARα/RARβ is a potentially therapeutic target for prevention of RIF. Full article

(This article belongs to the Section Biochemistry)

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20 pages, 1270 KiB

Open AccessArticle

Arabidopsis RIBA Proteins: Two out of Three Isoforms Have Lost Their Bifunctional Activity in Riboflavin Biosynthesis

by Hanna-Maija Hiltunen¹, Boris Illarionov², Boris Hedtke¹, Markus Fischer² and Bernhard Grimm^1,*

¹ Institute of Biology/Plant Physiology, Humboldt University Berlin, Philippstr. 13, Building 12, D-10115 Berlin, Germany

² Hamburg School of Food Science, Institute of Food Chemistry, University Hamburg, Grindelallee 117, 20146 Hamburg, Germany

Int. J. Mol. Sci. 2012, 13(11), 14086-14105; https://doi.org/10.3390/ijms131114086 - 31 Oct 2012

Cited by 24 | Viewed by 9653

Abstract

Riboflavin serves as a precursor for flavocoenzymes (FMN and FAD) and is essential for all living organisms. The two committed enzymatic steps of riboflavin biosynthesis are performed in plants by bifunctional RIBA enzymes comprised of GTP cyclohydrolase II (GCHII) and 3,4-dihydroxy-2-butanone-4-phosphate synthase (DHBPS). [...] Read more.

Riboflavin serves as a precursor for flavocoenzymes (FMN and FAD) and is essential for all living organisms. The two committed enzymatic steps of riboflavin biosynthesis are performed in plants by bifunctional RIBA enzymes comprised of GTP cyclohydrolase II (GCHII) and 3,4-dihydroxy-2-butanone-4-phosphate synthase (DHBPS). Angiosperms share a small RIBA gene family consisting of three members. A reduction of AtRIBA1 expression in the Arabidopsis rfd1mutant and in RIBA1 antisense lines is not complemented by the simultaneously expressed isoforms AtRIBA2 and AtRIBA3. The intensity of the bleaching leaf phenotype of RIBA1 deficient plants correlates with the inactivation of AtRIBA1 expression, while no significant effects on the mRNA abundance of AtRIBA2 and AtRIBA3 were observed. We examined reasons why both isoforms fail to sufficiently compensate for a lack of RIBA1 expression. All three RIBA isoforms are shown to be translocated into chloroplasts as GFP fusion proteins. Interestingly, both AtRIBA2 and AtRIBA3 have amino acid exchanges in conserved peptides domains that have been found to be essential for the two enzymatic functions. In vitro activity assays of GCHII and DHBPS with all of the three purified recombinant AtRIBA proteins and complementation of E. coli ribA and ribB mutants lacking DHBPS and GCHII expression, respectively, confirmed the loss of bifunctionality for AtRIBA2 and AtRIBA3. Phylogenetic analyses imply that the monofunctional, bipartite RIBA3 proteins, which have lost DHBPS activity, evolved early in tracheophyte evolution. Full article

(This article belongs to the Special Issue Flavins)

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9 pages, 267 KiB

Open AccessArticle

Detection of Glutamic Acid in Oilseed Rape Leaves Using Near Infrared Spectroscopy and the Least Squares-Support Vector Machine

by Yidan Bao^†, Wenwen Kong^†, Fei Liu^*

, Zhengjun Qiu and Yong He^*

¹ College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14106-14114; https://doi.org/10.3390/ijms131114106 - 31 Oct 2012

Cited by 11 | Viewed by 5588

Abstract

Amino acids are quite important indices to indicate the growth status of oilseed rape under herbicide stress. Near infrared (NIR) spectroscopy combined with chemometrics was applied for fast determination of glutamic acid in oilseed rape leaves. The optimal spectral preprocessing method was obtained [...] Read more.

Amino acids are quite important indices to indicate the growth status of oilseed rape under herbicide stress. Near infrared (NIR) spectroscopy combined with chemometrics was applied for fast determination of glutamic acid in oilseed rape leaves. The optimal spectral preprocessing method was obtained after comparing Savitzky-Golay smoothing, standard normal variate, multiplicative scatter correction, first and second derivatives, detrending and direct orthogonal signal correction. Linear and nonlinear calibration methods were developed, including partial least squares (PLS) and least squares-support vector machine (LS-SVM). The most effective wavelengths (EWs) were determined by the successive projections algorithm (SPA), and these wavelengths were used as the inputs of PLS and LS-SVM model. The best prediction results were achieved by SPA-LS-SVM (Raw) model with correlation coefficient r = 0.9943 and root mean squares error of prediction (RMSEP) = 0.0569 for prediction set. These results indicated that NIR spectroscopy combined with SPA-LS-SVM was feasible for the fast and effective detection of glutamic acid in oilseed rape leaves. The selected EWs could be used to develop spectral sensors, and the important and basic amino acid data were helpful to study the function mechanism of herbicide. Full article

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12 pages, 356 KiB

Open AccessArticle

Callose Deposition Is Required for Somatic Embryogenesis in Plasmolyzed Eleutherococcus senticosus Zygotic Embryos

by Lei Tao¹, Yang Yang², Qiuyu Wang¹ and Xiangling You^1,*

¹ College of Life Sciences, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China

² College of Landscape, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China

Int. J. Mol. Sci. 2012, 13(11), 14115-14126; https://doi.org/10.3390/ijms131114115 - 31 Oct 2012

Cited by 14 | Viewed by 7230

Abstract

Dynamic changes in callose content, which is deposited as a plant defense response to physiological changes, were analyzed during somatic embryogenesis in Eleutherococcus senticosus zygotic embryos plasmolyzed in 1.0 M mannitol. During plasmolysis, callose deposition was clearly observed inside the plasma membrane of [...] Read more.

Dynamic changes in callose content, which is deposited as a plant defense response to physiological changes, were analyzed during somatic embryogenesis in Eleutherococcus senticosus zygotic embryos plasmolyzed in 1.0 M mannitol. During plasmolysis, callose deposition was clearly observed inside the plasma membrane of zygotic embryo epidermal cells using confocal laser scanning microscopy. The callose content of zygotic embryos gradually increased between 0 and 12 h plasmolysis and remained stable after 24 h plasmolysis. During eight weeks induction of somatic embryogenesis, the callose content of explants plasmolyzed for 12 h was slightly higher than explants plasmolyzed for 6 or 24 h, with the largest differences observed after 6 weeks culture, which coincided with the maximum callose content and highest number of globular somatic embryos. The highest frequency of somatic embryo formation was observed in explants plasmolyzed for 12 h. The somatic embryo induction rate and number of somatic embryos per explant were markedly different in zygotic embryos pretreated with plasmolysis alone (78.0%, 43 embryos per explant) and those pretreated with plasmolysis and the callose synthase inhibitor 2-deoxy-d-glucose (11.5%, 8 embryos per explant). This study indicates that callose production is required for somatic embryogenesis in plasmolyzed explants. Full article

(This article belongs to the Section Biochemistry)

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Graphical abstract

9 pages, 197 KiB

Open AccessArticle

Bioavailability and Brain-Targeting of Geniposide in Gardenia-Borneol Co-Compound by Different Administration Routes in Mice

by Yang Lu, Shouying Du^*, Jie Bai, Pengyue Li, Ran Wen and Xuejiao Zhao

School of Chinese Materia Medica, Beijing University of Chinese Medicine, No. 6, Zhonghuan South Road, Wangjing, Chaoyang District, Beijing 100102, China

Int. J. Mol. Sci. 2012, 13(11), 14127-14135; https://doi.org/10.3390/ijms131114127 - 1 Nov 2012

Cited by 39 | Viewed by 6799

Abstract

Both geniposide (Ge) and borneol (Bo) are bioactive substances derived from traditional Chinese medicine. Injections containing co-compound of Gardenia-Borneol are widely used for stroke treatment in China, such as “Xingnaojing” multi-component injection. As more and more adverse reactions (especially drug allergy) were reported, [...] Read more.

Both geniposide (Ge) and borneol (Bo) are bioactive substances derived from traditional Chinese medicine. Injections containing co-compound of Gardenia-Borneol are widely used for stroke treatment in China, such as “Xingnaojing” multi-component injection. As more and more adverse reactions (especially drug allergy) were reported, it is urgent to find more effective and safer routes of administration for such kinds of medicines. In this paper, bioavailabilities and brain-target effects of geniposide in Gardenia-Borneol co-compound through different administration routes in mice were investigated. Geniposide concentrations in plasma and in brain of mice were determined by reversed-phase high-performance liquid chromatography. The pharmacokinetics parameters of intranasal (i.n.) and intragastric (i.g.) administration were compared with intravenous (i.v.) administration. The bioavailabilities of Ge were 85.38% and 28.76% for i.n. and i.g. while T_max were 1 min and 30 min. C_max were 21.881 ± 5.398, 1.914 ± 0.327 and 42.410 ± 6.268 μg/mL for i.n., i.g. and i.v., respectively. The AUC of Ge in brain were 32413.6 ± 4573.9, 6440.1 ± 863.7 and 37270.5 ± 4160.6 ng/g·min for i.n., i.g. and i.v., respectively. The drug target indexes (DTI) were 1.02 and 0.60 for i.n. and i.g. The results demonstrated that geniposide could be absorbed promptly and thoroughly by i.n. administration in mice and basically transported into the brain though blood vessel passways. Full article

(This article belongs to the Section Biochemistry)

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13 pages, 1459 KiB

Open AccessArticle

Preparation of Coaxial-Electrospun Poly[bis(p-methylphenoxy)]phosphazene Nanofiber Membrane for Enzyme Immobilization

by Shu-Gen Wang¹, Xin Jiang^1,2, Peng-Cheng Chen², An-Guo Yu² and Xiao-Jun Huang^2,*

¹ Key Laboratory of Eco-Textile, Ministry of Education, Jiangnan University, Wuxi 214122, China

² Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China

Int. J. Mol. Sci. 2012, 13(11), 14136-14148; https://doi.org/10.3390/ijms131114136 - 2 Nov 2012

Cited by 26 | Viewed by 7707

Abstract

A core/sheath nanofiber membrane with poly[bis(p-methylphenoxy)]phospha-zene (PMPPh) as the sheath and easily spinnable polyacrylonitrile (PAN) as the core was prepared via a coaxial electrospinning process. Field-emission scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the [...] Read more.

A core/sheath nanofiber membrane with poly[bis(p-methylphenoxy)]phospha-zene (PMPPh) as the sheath and easily spinnable polyacrylonitrile (PAN) as the core was prepared via a coaxial electrospinning process. Field-emission scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the nanofiber membrane. It was found that the concentration of the PAN spinning solution and the ratio of the core/sheath solution flow rates played a decisive role in the coaxial electrospinning process. In addition, the stabilized core/sheath PMPPh nanofiber membrane was investigated as a support for enzyme immobilization because of its excellent biocompatibility, high surface/volume ratio, and large porosity. Lipase from Candida rugosa was immobilized on the nanofiber membrane by adsorption. The properties of the immobilized lipase on the polyphosphazene nanofiber membrane were studied and compared with those of a PAN nanofiber membrane. The results showed that the adsorption capacity (20.4 ± 2.7 mg/g) and activity retention (63.7%) of the immobilized lipase on the polyphosphazene nanofiber membrane were higher than those on the PAN membrane. Full article

(This article belongs to the Special Issue Enzyme Optimization and Immobilization)

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9 pages, 462 KiB

Open AccessArticle

Construction of Mutant Glucose Oxidases with Increased Dye-Mediated Dehydrogenase Activity

by Yohei Horaguchi¹, Shoko Saito¹, Katsuhiro Kojima², Wakako Tsugawa¹, Stefano Ferri¹ and Koji Sode^1,2,*

¹ Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, Tokyo 184-8588, Japan

² Ultizyme International Ltd., 1-13-16, Minami, Meguro, Tokyo 152-0013, Japan

Int. J. Mol. Sci. 2012, 13(11), 14149-14157; https://doi.org/10.3390/ijms131114149 - 2 Nov 2012

Cited by 35 | Viewed by 8490

Abstract

Mutagenesis studies on glucose oxidases (GOxs) were conducted to construct GOxs with reduced oxidase activity and increased dehydrogenase activity. We focused on two representative GOxs, of which crystal structures have already been reported—Penicillium amagasakiense GOx (PDB ID; 1gpe) and Aspergillus niger GOx [...] Read more.

Mutagenesis studies on glucose oxidases (GOxs) were conducted to construct GOxs with reduced oxidase activity and increased dehydrogenase activity. We focused on two representative GOxs, of which crystal structures have already been reported—Penicillium amagasakiense GOx (PDB ID; 1gpe) and Aspergillus niger GOx (PDB ID; 1cf3). We constructed oxygen-interacting structural models for GOxs, and predicted the residues responsible for oxidative half reaction with oxygen on the basis of the crystal structure of cholesterol oxidase as well as on the fact that both enzymes are members of the glucose/methanol/choline (GMC) oxidoreductase family. Rational amino acid substitution resulted in the construction of an engineered GOx with drastically decreased oxidase activity and increased dehydrogenase activity, which was higher than that of the wild-type enzyme. As a result, the dehydrogenase/oxidase ratio of the engineered enzyme was more than 11-fold greater than that of the wild-type enzyme. These results indicate that alteration of the dehydrogenase/oxidase activity ratio of GOxs is possible by introducing a mutation into the putative functional residues responsible for oxidative half reaction with oxygen of these enzymes, resulting in a further increased dehydrogenase activity. This is the first study reporting the alteration of GOx electron acceptor preference from oxygen to an artificial electron acceptor. Full article

(This article belongs to the Special Issue Flavins)

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14 pages, 1060 KiB

Open AccessArticle

Apoptosis Induction of Human Prostate Carcinoma DU145 Cells by Diallyl Disulfide via Modulation of JNK and PI3K/AKT Signaling Pathways

by Dong Yeok Shin¹, Gi-Young Kim², Jun Hyuk Lee³, Byung Tae Choi⁴

, Young Hyun Yoo^5,* and Yung Hyun Choi⁶

¹ Dongnam Institute of Radiological & Medicine Sciences, Busan 619-953, Korea

² Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 690-756, Korea

³ Biotechnology Examination Division, Chemistry and Biotechnology Examination Bureau, Korean Intellectual Property Office, Daejeon 302-701, Korea

⁴ Division of Meridian and Structural Medicine, School of Korean Medicine, Pusan National University, Yangsan 626-870, Korea

⁵ Department of Anatomy and Cell Biology and Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan 602-714, Korea

⁶ Department of Biochemistry, College of Oriental Medicine and Anti-Aging Research Center, Dongeui University, Busan 614-052, Korea

Int. J. Mol. Sci. 2012, 13(11), 14158-14171; https://doi.org/10.3390/ijms131114158 - 2 Nov 2012

Cited by 48 | Viewed by 8801

Abstract

Diallyl disulfide (DADS), a sulfur compound derived from garlic, has various biological properties, such as anticancer, antiangiogenic and anti-inflammatory effects. However, the mechanisms of action underlying the compound's anticancer activity have not been fully elucidated. In this study, the apoptotic effects of DADS [...] Read more.

Diallyl disulfide (DADS), a sulfur compound derived from garlic, has various biological properties, such as anticancer, antiangiogenic and anti-inflammatory effects. However, the mechanisms of action underlying the compound's anticancer activity have not been fully elucidated. In this study, the apoptotic effects of DADS were investigated in DU145 human prostate carcinoma cells. Our results showed that DADS markedly inhibited the growth of the DU145 cells by induction of apoptosis. Apoptosis was accompanied by modulation of Bcl-2 and inhibitor of apoptosis protein (IAP) family proteins, depolarization of the mitochondrial membrane potential (MMP, ΔΨm) and proteolytic activation of caspases. We also found that the expression of death-receptor 4 (DR4) and Fas ligand (FasL) proteins was increased and that the level of intact Bid proteins was down-regulated by DADS. Moreover, treatment with DADS induced phosphorylation of mitogen-activated protein kinases (MAPKs), including extracellular-signal regulating kinase (ERK), p38 MAPK and c-Jun N-terminal kinase (JNK). A specific JNK inhibitor, SP600125, significantly blocked DADS-induced-apoptosis, whereas inhibitors of the ERK (PD98059) and p38 MAPK (SB203580) had no effect. The induction of apoptosis was also accompanied by inactivation of phosphatidylinositol 3-kinase (PI3K)/Akt and the PI3K inhibitor LY29004 significantly increased DADS-induced cell death. These findings provide evidence demonstrating that the proapoptotic effect of DADS is mediated through the activation of JNK and the inhibition of the PI3K/Akt signaling pathway in DU145 cells. Full article

(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2012)

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31 pages, 241 KiB

Open AccessArticle

The Innate Immune-Related Genes in Catfish

by Lei Gao^1,2,†, Chongbo He^1,2,*,†, Xueguang Liu³, Hao Su¹, Xianggang Gao^1,2, Yunfeng Li^1,2 and Weidong Liu^1,2

¹ Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China

² Key Laboratory of Marine Fishery Molecular Biology of Liaoning Province, Dalian 116023, China

³ Liaoning Fisheries Technology Extension Station, Shenyang 110031, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14172-14202; https://doi.org/10.3390/ijms131114172 - 2 Nov 2012

Cited by 33 | Viewed by 8706

Abstract

Catfish is one of the most important aquaculture species in America (as well as in Asia and Africa). In recent years, the production of catfish has suffered massive financial losses due to pathogen spread and breakouts. Innate immunity plays a crucial role in [...] Read more.

Catfish is one of the most important aquaculture species in America (as well as in Asia and Africa). In recent years, the production of catfish has suffered massive financial losses due to pathogen spread and breakouts. Innate immunity plays a crucial role in increasing resistance to pathogenic organisms and has generated increasing interest in the past few years. This review summarizes the current understanding of innate immune-related genes in catfish, including pattern recognition receptors, antimicrobial peptides, complements, lectins, cytokines, transferrin and gene expression profiling using microarrays and next generation sequencing technologies. This review will benefit the understanding of innate immune system in catfish and further efforts in studying the innate immune-related genes in fish. Full article

(This article belongs to the Section Biochemistry)

16 pages, 2564 KiB

Open AccessArticle

Effect of Superfine Grinding on Antidiabetic Activity of Bitter Melon Powder

by Ying Zhu, Ying Dong^*, Xiwen Qian, Fengjie Cui, Qin Guo, Xinghua Zhou, Yun Wang, Yi Zhang and Zhiyu Xiong

School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China

Int. J. Mol. Sci. 2012, 13(11), 14203-14218; https://doi.org/10.3390/ijms131114203 - 2 Nov 2012

Cited by 61 | Viewed by 9491

Abstract

The antidiabetic activities of bitter melon powders produced with lyophilization/superfine grinding and hot air drying/normal grinding were investigated in vivo for selecting a suitable bitter melon processing procedure. After a five-week treatment, bitter melon lyophilized superfine grinding powder (BLSP) had a higher antidiabetic [...] Read more.

The antidiabetic activities of bitter melon powders produced with lyophilization/superfine grinding and hot air drying/normal grinding were investigated in vivo for selecting a suitable bitter melon processing procedure. After a five-week treatment, bitter melon lyophilized superfine grinding powder (BLSP) had a higher antidiabetic activity with reducing fasting blood glucose levels from 21.40 to 12.54 mmol/L, the serum insulin levels from 40.93 to 30.74 mIU/L, and restoring activities of SOD compared with those in the bitter melon hot air drying powder (BAP) treated group. Furthermore, BLSP protected pancreatic tissues including islet beta cells and reduced the loss of islet cells. Combined with the difference of compositions in BLSP and BAP, it could be concluded that superfine grinding and lyophilization processes were beneficial for presenting the antidiabetic activity, which will provide a reference for direct utilization of bitter melon as a suitable functional food to relieve symptoms of diabetes. Full article

(This article belongs to the Section Biochemistry)

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24 pages, 6846 KiB

Open AccessReview

Noncanonical Reactions of Flavoenzymes

by Pablo Sobrado^1,2

¹ Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061, USA

² Virginia Tech Center for Drug Discovery, Virginia Tech, Blacksburg, VA 24061, USA

Int. J. Mol. Sci. 2012, 13(11), 14219-14242; https://doi.org/10.3390/ijms131114219 - 5 Nov 2012

Cited by 26 | Viewed by 11127

Abstract

Enzymes containing flavin cofactors are predominantly involved in redox reactions in numerous cellular processes where the protein environment modulates the chemical reactivity of the flavin to either transfer one or two electrons. Some flavoenzymes catalyze reactions with no net redox change. In these [...] Read more.

Enzymes containing flavin cofactors are predominantly involved in redox reactions in numerous cellular processes where the protein environment modulates the chemical reactivity of the flavin to either transfer one or two electrons. Some flavoenzymes catalyze reactions with no net redox change. In these reactions, the protein environment modulates the reactivity of the flavin to perform novel chemistries. Recent mechanistic and structural data supporting novel flavin functionalities in reactions catalyzed by chorismate synthase, type II isopentenyl diphosphate isomerase, UDP-galactopyranose mutase, and alkyl-dihydroxyacetonephosphate synthase are presented in this review. In these enzymes, the flavin plays either a direct role in acid/base reactions or as a nucleophile or electrophile. In addition, the flavin cofactor is proposed to function as a “molecular scaffold” in the formation of UDP-galactofuranose and alkyl-dihydroxyacetonephosphate by forming a covalent adduct with reaction intermediates. Full article

(This article belongs to the Special Issue Flavins)

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Graphical abstract

8 pages, 177 KiB

Open AccessArticle

Nuclear Microsatellite Primers for the Endangered Relict Fir, Abies pinsapo (Pinaceae) and Cross-Amplification in Related Mediterranean Species

by Jose M. Sánchez-Robles^1,*, Francisco Balao¹, Juan L. García-Castaño¹, Anass Terrab¹, Laura Navarro-Sampedro² and Salvador Talavera¹

¹ Departamento de Biología Vegetal y Ecología, Universidad de Sevilla, Apdo. 1095, Sevilla E-41080, Spain

² Centro de Investigación, Tecnología e Innovación (CITIUS), Universidad de Sevilla, Apdo. 1095, Sevilla E-41080, Spain

Int. J. Mol. Sci. 2012, 13(11), 14243-14250; https://doi.org/10.3390/ijms131114243 - 5 Nov 2012

Cited by 19 | Viewed by 6193

Abstract

Twelve nuclear microsatellite primers (nSSR) were developed for the endangered species Abies pinsapo Boiss. to enable the study of gene flow and genetic structure in the remaining distribution areas. Microsatellite primers were developed using next-generation sequencing (454) data from a single Abies pinsapo [...] Read more.

Twelve nuclear microsatellite primers (nSSR) were developed for the endangered species Abies pinsapo Boiss. to enable the study of gene flow and genetic structure in the remaining distribution areas. Microsatellite primers were developed using next-generation sequencing (454) data from a single Abies pinsapo individual. Primers were applied to thirty individuals from the three extant localities. The number of alleles per locus ranged from one to four. Cross-amplification was tested for other Abies species from the Mediterranean Basin, and most of the loci showed higher polymorphisms in the Mediterranean species than in A. pinsapo. These microsatellite markers provide tools for conservation genetic studies in Abies pinsapo as well other Abies species from the Mediterranean Basin. Full article

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Graphical abstract

11 pages, 258 KiB

Open AccessArticle

Effect of β-Cyclodextrin Complexation on Solubility and Enzymatic Conversion of Naringin

by Li Cui^1,2, Zhen-Hai Zhang¹, E Sun¹ and Xiao-Bin Jia^1,2,*

¹ Key Laboratory of New Drug Delivery System of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, 100 Shizi Road, Nanjing 210028, Jiangsu, China

² Nanjing University of Chinese Medicine, Nanjing 210046, Jiangsu, China

Int. J. Mol. Sci. 2012, 13(11), 14251-14261; https://doi.org/10.3390/ijms131114251 - 5 Nov 2012

Cited by 70 | Viewed by 8254

Abstract

In the present paper, the effect of β-cyclodextrin (β-CD) inclusion complexation on the solubility and enzymatic hydrolysis of naringin was investigated. The inclusion complex of naringin/β-CD at the molar ratio of 1:1 was obtained by the dropping method and was characterized by differential [...] Read more.

In the present paper, the effect of β-cyclodextrin (β-CD) inclusion complexation on the solubility and enzymatic hydrolysis of naringin was investigated. The inclusion complex of naringin/β-CD at the molar ratio of 1:1 was obtained by the dropping method and was characterized by differential scanning calorimetry. The solubility of naringin complexes in water at 37 ± 0.1 °C was 15 times greater than that of free naringin. Snailase-involved hydrolysis conditions were tested for the bioconversion of naringin into naringenin using the univariate experimental design. Naringin can be transformed into naringenin by snailase-involved hydrolysis. The optimum conditions for enzymatic hydrolysis were determined as follows: pH 5.0, temperature 37 °C, ratio of snailase/substrate 0.8, substrate concentration 20 mg·mL⁻¹, and reaction time 12 h. Under the optimum conditions, the transforming rate of naringenin from naringin for inclusion complexes and free naringin was 98.7% and 56.2% respectively, suggesting that β-CD complexation can improve the aqueous solubility and consequently the enzymatic hydrolysis rate of naringin. Full article

(This article belongs to the Section Biochemistry)

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16 pages, 382 KiB

Open AccessArticle

Chemical Composition and Antioxidant Activities of Three Polysaccharide Fractions from Pine Cones

by Ren-Bo Xu¹, Xin Yang^1,2,*, Jing Wang^2,3,*, Hai-Tian Zhao¹, Wei-Hong Lu¹, Jie Cui¹, Cui-Lin Cheng¹, Pan Zou¹, Wei-Wei Huang¹, Pu Wang¹, Wen-Jing Li¹ and Xing-Long Hu¹

¹ School of Food Science and Engineering, Harbin Institute of Technology, 73 Huanghe Road, Nangang District, Harbin 150090, China

² Key Laboratory of Agro-product Quality and Safety, Institute of Quality Standard & Testing Technology for Agro-Product, Chinese Academy of Agricultural Sciences, No.12 Zhongguancun South Street, Haidian District, Beijing 100081, China

³ Key Laboratory of Agrifood Safety and Quality, Ministry of Agriculture, No.12 Zhongguancun South Street, Haidian District, Beijing 100081, China

Int. J. Mol. Sci. 2012, 13(11), 14262-14277; https://doi.org/10.3390/ijms131114262 - 5 Nov 2012

Cited by 96 | Viewed by 9847

Abstract

The traditional method of gas chromatography-mass spectrometry for monosaccharide component analysis with pretreatment of acetylation is described with slight modifications and verified in detail in this paper. It was then successfully applied to the quantitative analysis of component monosaccharides in polysaccharides extracted from [...] Read more.

The traditional method of gas chromatography-mass spectrometry for monosaccharide component analysis with pretreatment of acetylation is described with slight modifications and verified in detail in this paper. It was then successfully applied to the quantitative analysis of component monosaccharides in polysaccharides extracted from the pine cones. The results demonstrated that the three pine cone polysaccharides all consisted of ribose, rhamnose, arabinose, xylose, mannose, glucose and galactose in different molar ratios. According to the recovery experiment, the described method was proved accurate and practical for the analysis of pine cone polysaccharides, meeting the need in the field of chemical analysis of Pinus plants. Furthermore; the chemical characteristics, such as neutral sugar, uronic acids, amino acids, molecular weights, and antioxidant activities of the polysaccharides were investigated by chemical and instrumental methods. The results showed that the chemical compositions of the polysaccharides differed from each other, especially in the content of neutral sugar and uronic acid. In the antioxidant assays, the polysaccharide fractions exhibited effective scavenging activities on ABTS radical and hydroxyl radical, with their antioxidant capabilities decreasing in the order of PKP > PAP > PSP. Therefore, although the polysaccharide fractions had little effect on superoxide radical scavenging, they still have potential to be developed as natural antioxidant agents in functional foods or medicine. Full article

(This article belongs to the Section Biochemistry)

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16 pages, 1076 KiB

Open AccessArticle

Molecular Signaling Involved in Oxysterol-Induced β₁-Integrin Over-Expression in Human Macrophages

by Simona Gargiulo^†, Paola Gamba^†, Gabriella Testa

, Barbara Sottero, Marco Maina, Tina Guina, Fiorella Biasi, Giuseppe Poli^* and Gabriella Leonarduzzi

¹ Department of Clinical and Biological Sciences, University of Turin, San Luigi Hospital, Regione Gonzole 10, Orbassano 10043, Turin, Italy

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14278-14293; https://doi.org/10.3390/ijms131114278 - 5 Nov 2012

Cited by 16 | Viewed by 5963

Abstract

The hypercholesterolemia-atherosclerosis association is now established; hypercholesterolemia may induce vascular-cell activation, subsequently increasing expression of adhesion molecules, cytokines, chemokines, growth factors, and other key inflammatory molecules. Among inflammatory molecules expressed by vascular cells, integrins play a critical role in regulating macrophage activation and [...] Read more.

The hypercholesterolemia-atherosclerosis association is now established; hypercholesterolemia may induce vascular-cell activation, subsequently increasing expression of adhesion molecules, cytokines, chemokines, growth factors, and other key inflammatory molecules. Among inflammatory molecules expressed by vascular cells, integrins play a critical role in regulating macrophage activation and migration to the site of inflammation, by mediating cell-cell and cell-extracellular matrix interactions. The main lipid oxidation products present in oxidized LDL that may be responsible for inflammatory processes in atherogenesis, are cholesterol oxidation products, known as oxysterols. This study demonstrates the effect of an oxysterol mixture, compatible with that detectable in human hypercholesterolemic plasma, on the expression and synthesis of β₁-integrin in cells of the macrophage lineage. The molecular signaling whereby oxysterols induce β₁-integrin up-regulation is also comprehensively investigated. Over-expression of β₁-integrin depends on activation of classic and novel members of protein kinase C and extracellular signal-regulated kinases 1 and 2, as well as of the up-stream G-protein (Gq and G13), c-Src, and phospholipase C. In addition, the localization of β₁-integrin in advanced human carotid plaques is highlighted, marking its importance in atherosclerotic plaque progression. Full article

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17 pages, 1108 KiB

Open AccessArticle

Content and Color Stability of Anthocyanins Isolated from Schisandra chinensis Fruit

by Chunhui Ma, Lei Yang^*, Fengjian Yang, Wenjie Wang, Chunjian Zhao

and Yuangang Zu^*

State Engineering Laboratory for Bioresource Eco-Utilization, Northeast Forestry University, Harbin 150040, China

Int. J. Mol. Sci. 2012, 13(11), 14294-14310; https://doi.org/10.3390/ijms131114294 - 5 Nov 2012

Cited by 38 | Viewed by 8239

Abstract

In this work, a multivariate study based on Box-Behnken Design was used to evaluate the influence of three major variables affecting the performance of the extraction process of Schisandra chinensis anthocyanins. The optimum parameters were 5.5 h extraction time; 1:19 solid-liquid ratio and [...] Read more.

In this work, a multivariate study based on Box-Behnken Design was used to evaluate the influence of three major variables affecting the performance of the extraction process of Schisandra chinensis anthocyanins. The optimum parameters were 5.5 h extraction time; 1:19 solid-liquid ratio and 260 r/min stirring rate, respectively. The extraction yield of anthocyanins was 29.06 mg/g under the optimum conditions. Moreover, many factors on the impact of heating, ultrasound, microwave treatment and ultraviolet irradiation on content and color stability of anthocyanins from Schisandra chinensis fruit were investigated. The results show that thermal degradation reaction of anthocyanins complies with the first order reaction kinetics, and the correlation coefficient is greater than 0.9950 at 40–80 °C. Ultrasound and microwave treatment has little effect on the stability of anthocyanins, and the extraction time of ultrasound and microwave should be no more than 60 min and 5 min, respectively. The anthocyanins degradation effect of UVC ultraviolet radiation is greater than UVA and UVB; after 9 h ultraviolet radiation, the anthocyanins content degradation of UVC is 23.9 ± 0.7%, and the ΔE* was changed from 62.81 to 76.52 ± 2.3. Through LC-MS analysis, the major composition of Schisandra chinensis anthocyanins was cyanidin-3-O-xylosylrutinoside. Full article

(This article belongs to the Section Biochemistry)

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15 pages, 1528 KiB

Open AccessArticle

Protection against Ischemia-Induced Oxidative Stress Conferred by Vagal Stimulation in the Rat Heart: Involvement of the AMPK-PKC Pathway

by Shan-Shan Kong, Jin-Jun Liu, Xiao-Jiang Yu, Yi Lu and Wei-Jin Zang^*

Department of Pharmacology, College of Medicine, Xi'an Jiaotong University, Xi'an 710061, China

Int. J. Mol. Sci. 2012, 13(11), 14311-14325; https://doi.org/10.3390/ijms131114311 - 5 Nov 2012

Cited by 52 | Viewed by 8588

Abstract

Reactive oxygen species (ROS) production is an important mechanism in myocardial ischemia and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is one of major sources of ROS in the heart. Previous studies showed that vagus nerve stimulation (VNS) is beneficial in treating ischemic heart [...] Read more.

Reactive oxygen species (ROS) production is an important mechanism in myocardial ischemia and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is one of major sources of ROS in the heart. Previous studies showed that vagus nerve stimulation (VNS) is beneficial in treating ischemic heart diseases. However, the effect of VNS on ROS production remains elusive. In this study, we investigated the role of VNS onischemia-induced ROS production. Our results demonstrated that VNS alleviated the myocardial injury, attenuated the cardiac dysfunction, reserved the antioxidant enzyme activity and inhibited the formation of ROS as evidenced by the decreased NADPH oxidase (Nox) activity and superoxide fluorescence intensity as well as the expression of p67phox, Rac1 and nitrotyrosine. Furthermore, VNS resulted in the phosphorylation and activation of adenosine monophosphate activated protein kinase (AMPK), which in turn led to an inactivation of Nox by protein kinase C (PKC); however, the phenomena were repressed by the administration of a muscarinic antagonist atropine. Taken together, these data indicate that VNS decreases ROS via AMPK-PKC-Nox pathway; this may have potential importance for the treatment of ischemic heart diseases. Full article

(This article belongs to the Special Issue Advances in Free Radicals in Biology and Medicine)

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18 pages, 1721 KiB

Open AccessArticle

Molecular Mechanisms of Pharmaceutical Drug Binding into Calsequestrin

by Arun K. Subra¹, Mark S. Nissen¹, Kevin M. Lewis¹, Ashwin K. Muralidharan¹, Emiliano J. Sanchez², Hendrik Milting³ and ChulHee Kang^1,2,*

¹ Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA

² School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USA

³ Herz- und Diabeteszentrum NRW, Klinik der Ruhr Universitaet Bochum, Erich and Hanna Klessmann-Institut für Kardiovaskuläre Forschung und Entwicklung, 32545 Bad Oeynhausen, Germany

Int. J. Mol. Sci. 2012, 13(11), 14326-14343; https://doi.org/10.3390/ijms131114326 - 6 Nov 2012

Cited by 8 | Viewed by 7397

Abstract

Calsequestrin (CASQ) is a major Ca²⁺-storage/buffer protein present in the sarcoplasmic reticulum of both skeletal (CASQ1) and cardiac (CASQ2) muscles. CASQ has significant affinity for a number of pharmaceutical drugs with known muscular toxicities. Our approach, with in silico molecular docking, [...] Read more.

Calsequestrin (CASQ) is a major Ca²⁺-storage/buffer protein present in the sarcoplasmic reticulum of both skeletal (CASQ1) and cardiac (CASQ2) muscles. CASQ has significant affinity for a number of pharmaceutical drugs with known muscular toxicities. Our approach, with in silico molecular docking, single crystal X-ray diffraction, and isothermal titration calorimetry (ITC), identified three distinct binding pockets on the surface of CASQ2, which overlap with 2-methyl-2,4-pentanediol (MPD) binding sites observed in the crystal structure. Those three receptor sites based on canine CASQ1 crystal structure gave a high correlation (R² = 0.80) to our ITC data. Daunomycin, doxorubicin, thioridazine, and trifluoperazine showed strong affinity to the S1 site, which is a central cavity formed between three domains of CASQ2. Some of the moderate-affinity drugs and some high-affinity drugs like amlodipine and verapamil displayed their binding into S2 sites, which are the thioredoxin-like fold present in each CASQ domain. Docking predictions combined with dissociation constants imply that presence of large aromatic cores and less flexible functional groups determines the strength of binding affinity to CASQ. In addition, the predicted binding pockets for both caffeine and epigallocatechin overlapped with the S1 and S2 sites, suggesting competitive inhibition by these natural compounds as a plausible explanation for their antagonistic effects on cardiotoxic side effects. Full article

(This article belongs to the Section Biochemistry)

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12 pages, 400 KiB

Open AccessArticle

Selection of Suitable Reference Genes for Normalization of Quantitative Real-Time PCR in Cartilage Tissue Injury and Repair in Rabbits

by Xiao-Xiang Peng^1,†, Rong-Lan Zhao^1,†, Wei Song¹, Hai-Rong Chu¹, Meng Li¹, Shu-Ya Song¹, Guang-Zhou Li¹ and Dong-Chun Liang^2,3,*

¹ Shandong Provincial Key Laboratory of Clinical Laboratory Diagnostics, Department of Medical Laboratory, Weifang Medical University, Shandong 261053, China

² Institute of Endocrinology and Metabolic Disease Hospital, Tianjin Medical University, Tianjin 300070, China

³ Key Lab of Hormone and Development, Tianjin Ministry, Tianjin 300070, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14344-14355; https://doi.org/10.3390/ijms131114344 - 6 Nov 2012

Cited by 21 | Viewed by 6091

Abstract

When studying the altered expression of genes associated with cartilage regeneration by quantitative real-time RT-PCR (RT-qPCR), reference genes with highly stable expression during different stages of chondrocyte developmental are necessary to normalize gene expression accurately. Until now, no reports evaluating expression changes of [...] Read more.

When studying the altered expression of genes associated with cartilage regeneration by quantitative real-time RT-PCR (RT-qPCR), reference genes with highly stable expression during different stages of chondrocyte developmental are necessary to normalize gene expression accurately. Until now, no reports evaluating expression changes of commonly used reference genes in rabbit articular cartilage have been published. In this study, defects were made in rabbit articular cartilage, with or without insulin-like growth factor 1 (IGF-1) treatment, to create different chondrocyte living environments. The stability and intensity of the expressions of the candidate reference genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S Ribosomal RNA (18S rRNA), cyclophilin (CYP), hypoxanthine phosphoribosyl transferase (HPRT1), and β-2-microglobulin (B2M) were evaluated. The data were analyzed by geNorm and NormFinder. B2M and 18S rRNA were identified to be suitable reference genes for rabbit cartilage tissues. Full article

(This article belongs to the Section Biochemistry)

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29 pages, 16474 KiB

Open AccessArticle

Electronic Structure and Phase Transition in Ferroelectic Sn2P2S6 Crystal

by Konstantin Glukhov^1,*, Kristina Fedyo¹, Juras Banys² and Yulian Vysochanskii¹

¹ Institute for Solid State Physics and Chemistry, Uzhgorod National University, Voloshyn Street 54, Uzhgorod 88000, Ukraine

² Faculty of Physics, Vilnius University, LT-10222 Vilnius, Lithuania

Int. J. Mol. Sci. 2012, 13(11), 14356-14384; https://doi.org/10.3390/ijms131114356 - 6 Nov 2012

Cited by 42 | Viewed by 8314

Abstract

An analysis of the P2S6 cluster electronic structure and its comparison with the crystal valence band in the paraelectric and ferroelectric phases has been done by first-principles calculations for Sn2P2S6 ferroelectrics. The origin of ferroelectricity has been outlined. It was established that the [...] Read more.

An analysis of the P2S6 cluster electronic structure and its comparison with the crystal valence band in the paraelectric and ferroelectric phases has been done by first-principles calculations for Sn2P2S6 ferroelectrics. The origin of ferroelectricity has been outlined. It was established that the spontaneous polarization follows from the stereochemical activity of the electron lone pair of tin cations, which is determined by hybridization with P2S6 molecular orbitals. The chemical bonds covalence increase and rearrangement are related to the valence band changes at transition from the paraelectric phase to the ferroelectric phase. Full article

(This article belongs to the Special Issue Molecular Symmetry)

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16 pages, 1689 KiB

Open AccessReview

Monitoring Biosensor Activity in Living Cells with Fluorescence Lifetime Imaging Microscopy

by Julia M. Hum, Amanda P. Siegel, Fredrick M. Pavalko and Richard N. Day^*

Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 635 Barnhill Dr., MS 333, Indianapolis, IN 46202, USA

Int. J. Mol. Sci. 2012, 13(11), 14385-14400; https://doi.org/10.3390/ijms131114385 - 7 Nov 2012

Cited by 22 | Viewed by 9884

Abstract

Live-cell microscopy is now routinely used to monitor the activities of the genetically encoded biosensor proteins that are designed to directly measure specific cell signaling events inside cells, tissues, or organisms. Most fluorescent biosensor proteins rely on Förster resonance energy transfer (FRET) to [...] Read more.

Live-cell microscopy is now routinely used to monitor the activities of the genetically encoded biosensor proteins that are designed to directly measure specific cell signaling events inside cells, tissues, or organisms. Most fluorescent biosensor proteins rely on Förster resonance energy transfer (FRET) to report conformational changes in the protein that occur in response to signaling events, and this is commonly measured with intensity-based ratiometric imaging methods. An alternative method for monitoring the activities of the FRET-based biosensor proteins is fluorescence lifetime imaging microscopy (FLIM). FLIM measurements are made in the time domain, and are not affected by factors that commonly limit intensity measurements. In this review, we describe the use of the digital frequency domain (FD) FLIM method for the analysis of FRET signals. We illustrate the methods necessary for the calibration of the FD FLIM system, and demonstrate the analysis of data obtained from cells expressing “FRET standard” fusion proteins. We then use the FLIM-FRET approach to monitor the changes in activities of two different biosensor proteins in specific regions of single living cells. Importantly, the factors required for the accurate determination and reproducibility of lifetime measurements are described in detail. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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20 pages, 584 KiB

Open AccessArticle

Changes on the Caco-2 Secretome through Differentiation Analyzed by 2-D Differential In-Gel Electrophoresis (DIGE)

by Andrés García-Lorenzo, Ana M. Rodríguez-Piñeiro^†, Francisco J. Rodríguez-Berrocal, María Páez de la Cadena and Vicenta S. Martínez-Zorzano^*

¹ Department of Biochemistry, Genetics and Immunology, Faculty of Biology, University of Vigo, 36310 Vigo, Spain

^† Present address: Institute of Biomedicine, University of Gothenburg, 41390 Gothenburg, Sweden

Int. J. Mol. Sci. 2012, 13(11), 14401-14420; https://doi.org/10.3390/ijms131114401 - 7 Nov 2012

Cited by 18 | Viewed by 8849

Abstract

Colorectal cancer is still a major health burden worldwide, and its diagnosis has not improved in recent years due to a lack of appropriate diagnostic serum markers. Aiming to find new diagnostic proteins, we applied the proteomic DIGE technology to analyze changes in [...] Read more.

Colorectal cancer is still a major health burden worldwide, and its diagnosis has not improved in recent years due to a lack of appropriate diagnostic serum markers. Aiming to find new diagnostic proteins, we applied the proteomic DIGE technology to analyze changes in the secretome before/after differentiation of the colon adenocarcinoma Caco-2 cell line, an accepted in vitro model to study colorectal tumorigenesis. When the secretomes from undifferentiated (tumor-like) and differentiated cells (resembling healthy enterocytes) were compared, we found 96 spots differentially expressed. After MS/MS analysis, 22 spots corresponding to 15 different proteins were identified. Principal component analysis demonstrated these 22 spots could serve as a discriminatory panel between the tumor-like and normal-like cells. Among the identified proteins, the translationally-controlled tumor protein (TCTP), the transforming growth factor-beta-induced protein ig-h3 (TGFβIp), and the Niemann-Pick disease type C2 protein (NPC2) are interesting candidates for future studies focused on their utility as serum biomarkers of colorectal cancer. Full article

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

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13 pages, 1356 KiB

Open AccessArticle

Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

by Xiaoyun Chen^1,†, Xiaofu Wang^1,†, Nuo Jin², Yu Zhou¹, Sainan Huang^1,3, Qingmei Miao¹, Qing Zhu¹ and Junfeng Xu^1,*

¹ Institute of Agriculture Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

² School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China

³ College of Chemistry and Life Science, Shenyang Normal University, Shenyang 110034, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14421-14433; https://doi.org/10.3390/ijms131114421 - 7 Nov 2012

Cited by 64 | Viewed by 9029

Abstract

Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common [...] Read more.

Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%–0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops. Full article

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12 pages, 2882 KiB

Open AccessArticle

Facile Synthesis of Calcium Borate Nanoparticles and the Annealing Effect on Their Structure and Size

by Maryam Erfani^1,*, Elias Saion¹, Nayereh Soltani¹

, Mansor Hashim¹, Wan Saffiey B. Wan Abdullah² and Manizheh Navasery¹

¹ Department of Physics, Faculty of Science, University Putra Malaysia, 43400UPM Serdang, Selangor, Malaysia

² Department of Non-ionizing Radiation, Malaysian Nuclear Agency (Nuclear Malaysia), Bangi, 43000 Kajang, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 14434-14445; https://doi.org/10.3390/ijms131114434 - 8 Nov 2012

Cited by 40 | Viewed by 8756

Abstract

Calcium borate nanoparticles have been synthesized by a thermal treatment method via facile co-precipitation. Differences of annealing temperature and annealing time and their effects on crystal structure, particle size, size distribution and thermal stability of nanoparticles were investigated. The formation of calcium borate [...] Read more.

Calcium borate nanoparticles have been synthesized by a thermal treatment method via facile co-precipitation. Differences of annealing temperature and annealing time and their effects on crystal structure, particle size, size distribution and thermal stability of nanoparticles were investigated. The formation of calcium borate compound was characterized by X-ray diffraction (XRD) and Fourier Transform Infrared spectroscopy (FTIR), Transmission electron microscopy (TEM), and Thermogravimetry (TGA). The XRD patterns revealed that the co-precipitated samples annealed at 700 °C for 3 h annealing time formed an amorphous structure and the transformation into a crystalline structure only occurred after 5 h annealing time. It was found that the samples annealed at 900 °C are mostly metaborate (CaB₂O₄) nanoparticles and tetraborate (CaB₄O₇) nanoparticles only observed at 970 °C, which was confirmed by FTIR. The TEM images indicated that with increasing the annealing time and temperature, the average particle size increases. TGA analysis confirmed the thermal stability of the annealed samples at higher temperatures. Full article

(This article belongs to the Section Materials Science)

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5 pages, 163 KiB

Open AccessArticle

Development of New Polymorphic Microsatellite Loci for the Barley Stem Gall Midge, Mayetiola hordei (Diptera: Cecidomyiidae) from an Enriched Library

by Maha Mezghani-Khemakhem^1,*, Dhia Bouktila^1,2, Nathalie Casse³, Houcine Maaroufi¹, Mohamed Makni¹ and Hanem Makni^1,4

¹ Genomics of Crop Insect Pests, Faculté des Sciences de Tunis, University of Tunis El-Manar, El Manar 2092, Tunisia

² Higher Institute of Biotechnology of Béja, University of Jendouba, Avenue Habib Bourguiba, Béja 9000, Tunisia

³ Laboratory Mer, Molécules, Santé (MMS), UFR Sciences and Techniques Le Mans, University of Maine, Le Mans 72085, France

⁴ Higher Institute of Animation for Youth and Culture, University of Tunis, Bir El Bey 2055, Tunisia

Int. J. Mol. Sci. 2012, 13(11), 14446-14450; https://doi.org/10.3390/ijms131114446 - 8 Nov 2012

Cited by 6 | Viewed by 5243

Abstract

Using an enriched library method, seven polymorphic microsatellite loci were isolated from the barley stem gall midge, Mayetiola hordei. Polymorphism at loci was surveyed on 57 individual midges collected on barley in Tunisia. Across loci, polymorphism ranged from two to six alleles [...] Read more.

Using an enriched library method, seven polymorphic microsatellite loci were isolated from the barley stem gall midge, Mayetiola hordei. Polymorphism at loci was surveyed on 57 individual midges collected on barley in Tunisia. Across loci, polymorphism ranged from two to six alleles per locus. The observed heterozygosity varied between 0.070 and 0.877. Based on the number of alleles detected and the associated levels of heterozygosity, we believe that these loci will prove useful for population genetic studies on M. hordei. Full article

(This article belongs to the Section Biochemistry)

19 pages, 1132 KiB

Open AccessArticle

A Generic Force Field for Protein Coarse-Grained Molecular Dynamics Simulation

by Junfeng Gu¹, Fang Bai², Honglin Li³ and Xicheng Wang^1,*

¹ State Key Laboratory of Structural Analysis for Industrial Equipment, Department of Engineering Mechanics, Dalian University of Technology, Dalian 116023, China

² Faculty of Chemical, Environmental and Biological Science and Technology, Dalian University of Technology, Dalian 116023, China

³ School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China

Int. J. Mol. Sci. 2012, 13(11), 14451-14469; https://doi.org/10.3390/ijms131114451 - 8 Nov 2012

Cited by 19 | Viewed by 8737

Abstract

Coarse-grained (CG) force fields have become promising tools for studies of protein behavior, but the balance of speed and accuracy is still a challenge in the research of protein coarse graining methodology. In this work, 20 CG beads have been designed based on [...] Read more.

Coarse-grained (CG) force fields have become promising tools for studies of protein behavior, but the balance of speed and accuracy is still a challenge in the research of protein coarse graining methodology. In this work, 20 CG beads have been designed based on the structures of amino acid residues, with which an amino acid can be represented by one or two beads, and a CG solvent model with five water molecules was adopted to ensure the consistence with the protein CG beads. The internal interactions in protein were classified according to the types of the interacting CG beads, and adequate potential functions were chosen and systematically parameterized to fit the energy distributions. The proposed CG force field has been tested on eight proteins, and each protein was simulated for 1000 ns. Even without any extra structure knowledge of the simulated proteins, the Cα root mean square deviations (RMSDs) with respect to their experimental structures are close to those of relatively short time all atom molecular dynamics simulations. However, our coarse grained force field will require further refinement to improve agreement with and persistence of native-like structures. In addition, the root mean square fluctuations (RMSFs) relative to the average structures derived from the simulations show that the conformational fluctuations of the proteins can be sampled. Full article

(This article belongs to the Section Biochemistry)

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22 pages, 376 KiB

Open AccessReview

Experimental and Therapeutic Opportunities for Stem Cells in Multiple Sclerosis

by Rickie Patani^1,2 and Siddharthan Chandran^3,*

¹ Anne McLaren Laboratory for Regenerative Medicine, Stem Cell Institute, University of Cambridge, Forvie Site, Robinson Way, Cambridge CB2 0SZ, UK

² Cambridge Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Forvie Site, Robinson Way, Cambridge CB2 0PY, UK

³ Euan MacDonald Centre, University of Edinburgh, Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK

Int. J. Mol. Sci. 2012, 13(11), 14470-14491; https://doi.org/10.3390/ijms131114470 - 8 Nov 2012

Cited by 6 | Viewed by 6807

Abstract

Multiple Sclerosis (MS) is an inflammatory demyelinating neurodegenerative disorder of the brain and spinal cord that causes significant disability in young adults. Although the precise aetiopathogenesis of MS remains unresolved, its pathological hallmarks include inflammation, demyelination, axonal injury (acute and chronic), astrogliosis and [...] Read more.

Multiple Sclerosis (MS) is an inflammatory demyelinating neurodegenerative disorder of the brain and spinal cord that causes significant disability in young adults. Although the precise aetiopathogenesis of MS remains unresolved, its pathological hallmarks include inflammation, demyelination, axonal injury (acute and chronic), astrogliosis and variable remyelination. Despite major recent advances in therapeutics for the early stage of the disease there are currently no disease modifying treatments for the progressive stage of disease, whose pathological substrate is axonal degeneration. This represents the great and unmet clinical need in MS. Against this background, human stem cells offer promise both to improve understanding of disease mechanism(s) through in-vitro modeling as well as potentially direct use to supplement and promote remyelination, an endogenous reparative process where entire myelin sheaths are restored to demyelinated axons. Conceptually, stem cells can act directly to myelinate axons or indirectly through different mechanisms to promote endogenous repair; importantly these two mechanisms of action are not mutually exclusive. We propose that discovery of novel methods to invoke or enhance remyelination in MS may be the most effective therapeutic strategy to limit axonal damage and instigate restoration of structure and function in this debilitating condition. Human stem cell derived neurons and glia, including patient specific cells derived through reprogramming, provide an unprecedented experimental system to model MS “in a dish” as well as enable high-throughput drug discovery. Finally, we speculate upon the potential role for stem cell based therapies in MS. Full article

(This article belongs to the Special Issue Recent Advances in the Research of Multiple Sclerosis)

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26 pages, 1522 KiB

Open AccessArticle

Role of Key Residues at the Flavin Mononucleotide (FMN):Adenylyltransferase Catalytic Site of the Bifunctional Riboflavin Kinase/Flavin Adenine Dinucleotide (FAD) Synthetase from Corynebacterium ammoniagenes

by Ana Serrano¹, Susana Frago¹, Adrián Velázquez-Campoy^1,2 and Milagros Medina^1,*

¹ Department of Biochemistry and Cellular and Molecular Biology, Faculty of Sciences, Institute for Biocomputation and Physics of Complex Systems (BIFI)-Joint Unit BIFI-IQFR (CSIC), University of Zaragoza, Pedro Cerbuna, 12. 50009 Zaragoza, Spain

² Foundation, Aragonian Government, 50018 Zaragoza, Spain

Int. J. Mol. Sci. 2012, 13(11), 14492-14517; https://doi.org/10.3390/ijms131114492 - 8 Nov 2012

Cited by 30 | Viewed by 10180

Abstract

In mammals and in yeast the conversion of Riboflavin (RF) into flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) is catalysed by the sequential action of two enzymes: an ATP:riboflavin kinase (RFK) and an ATP:FMN adenylyltransferase (FMNAT). However, most prokaryotes depend on a [...] Read more.

In mammals and in yeast the conversion of Riboflavin (RF) into flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) is catalysed by the sequential action of two enzymes: an ATP:riboflavin kinase (RFK) and an ATP:FMN adenylyltransferase (FMNAT). However, most prokaryotes depend on a single bifunctional enzyme, FAD synthetase (FADS), which folds into two modules: the C-terminal associated with RFK activity and the N-terminal associated with FMNAT activity. Sequence and structural analysis suggest that the 28-HxGH-31, 123-Gx(D/N)-125 and 161-xxSSTxxR-168 motifs from FADS must be involved in ATP stabilisation for the adenylylation of FMN, as well as in FAD stabilisation for FAD phyrophosphorolysis. Mutants were produced at these motifs in the Corynebacterium ammoniagenes FADS (CaFADS). Their effects on the kinetic parameters of CaFADS activities (RFK, FMNAT and FAD pyrophosphorilase), and on substrates and product binding properties indicate that H28, H31, N125 and S164 contribute to the geometry of the catalytically competent complexes at the FMNAT-module of CaFADS. Full article

(This article belongs to the Special Issue Flavins)

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Graphical abstract

15 pages, 985 KiB

Open AccessArticle

Protein Profiling of Blood Samples from Patients with Hereditary Leiomyomatosis and Renal Cell Cancer by Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

by Takao Kamai^1,*, Naohisa Tomosugi², Hideyuki Abe¹, Yasushi Kaji³, Tetsunari Oyama⁴ and Ken-Ichiro Yoshida¹

¹ Department of Urology, Dokkyo Medical University, Tochigi 321-0293, Japan

² Proteomics Research Unit, Division of Advanced Medicine, Medical Research Institute, Kanazawa Medical University, Ishikawa 920-0293, Japan

³ Department of Radiology, Dokkyo Medical University, Tochigi 321-0293, Japan

⁴ Department of Pathology, Dokkyo Medical University, Tochigi 321-0293, Japan

Int. J. Mol. Sci. 2012, 13(11), 14518-14532; https://doi.org/10.3390/ijms131114518 - 8 Nov 2012

Cited by 12 | Viewed by 7038

Abstract

Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an extremely rare syndrome with autosomal dominant inheritance. HLRCC is characterized by a predisposition to leiomyomas of the skin and the uterus as well as renal cell carcinoma. The disease-related gene has been identified as [...] Read more.

Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an extremely rare syndrome with autosomal dominant inheritance. HLRCC is characterized by a predisposition to leiomyomas of the skin and the uterus as well as renal cell carcinoma. The disease-related gene has been identified as fumarate hydratase (fumarase, FH), which encodes an enzyme involved in the mitochondrial tricarboxylic acid cycle. Protein profiling may give some insight into the molecular pathways of HLRCC. Therefore, we performed protein profiling of blood samples from HLRCC patients, their family members, and healthy volunteers, using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) coupled with IMAC-Cu chips. For hierarchical clustering analysis, we used the 45 peaks that revealed significant differences in single-marker analysis over the range from 1500 to 15,000 m/z. Heat map analysis based on the results of clustering distinguished the HLRCC kindred from non-HLRCC subjects with a sensitivity of 94% and a specificity of 90%. SELDI-TOF MS profiling of blood samples can be applied to identify patients with HLRCC and to assess specific molecular mechanisms involved in this condition. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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12 pages, 477 KiB

Open AccessArticle

Optimization of Pressurized Liquid Extraction of Three Major Acetophenones from Cynanchum bungei Using a Box-Behnken Design

by Wei Li^1,†

, Li-Chun Zhao^2,†, Yin-Shi Sun³, Feng-Jie Lei¹, Zi Wang¹, Xiong-Bin Gui^2,* and Hui Wang^4,*

¹ College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun 130118, China

² The Affiliated Ruikang Hospital, Guangxi University of Chinese Medicine, Nanning 530011, China

³ College of Agronomy, Shandong Agricultural University, Taian 271018, China

⁴ China-Japan Union Hospital, Jilin University, Changchun 130033, China

^† These two authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14533-14544; https://doi.org/10.3390/ijms131114533 - 8 Nov 2012

Cited by 19 | Viewed by 7058

Abstract

In this work, pressurized liquid extraction (PLE) of three acetophenones (4-hydroxyacetophenone, baishouwubenzophenone, and 2,4-dihydroxyacetophenone) from Cynanchum bungei (ACB) were investigated. The optimal conditions for extraction of ACB were obtained using a Box-Behnken design, consisting of 17 experimental points, as follows: Ethanol (100%) as [...] Read more.

In this work, pressurized liquid extraction (PLE) of three acetophenones (4-hydroxyacetophenone, baishouwubenzophenone, and 2,4-dihydroxyacetophenone) from Cynanchum bungei (ACB) were investigated. The optimal conditions for extraction of ACB were obtained using a Box-Behnken design, consisting of 17 experimental points, as follows: Ethanol (100%) as the extraction solvent at a temperature of 120 °C and an extraction pressure of 1500 psi, using one extraction cycle with a static extraction time of 17 min. The extracted samples were analyzed by high-performance liquid chromatography using an UV detector. Under this optimal condition, the experimental values agreed with the predicted values by analysis of variance. The ACB extraction yield with optimal PLE was higher than that obtained by soxhlet extraction and heat-reflux extraction methods. The results suggest that the PLE method provides a good alternative for acetophenone extraction. Full article

(This article belongs to the Section Biochemistry)

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20 pages, 427 KiB

Open AccessArticle

Lateral Distribution of NBD-PC Fluorescent Lipid Analogs in Membranes Probed by Molecular Dynamics-Assisted Analysis of Förster Resonance Energy Transfer (FRET) and Fluorescence Quenching

by Luís M. S. Loura^1,2

¹ Faculty of Pharmacy, University of Coimbra, Health Sciences Campus, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal

² Centre for Chemistry-Coimbra, Rua Larga, 3004-535 Coimbra, Portugal

Int. J. Mol. Sci. 2012, 13(11), 14545-14564; https://doi.org/10.3390/ijms131114545 - 8 Nov 2012

Cited by 21 | Viewed by 8425

Abstract

Förster resonance energy transfer (FRET) is a powerful tool used for many problems in membrane biophysics, including characterization of the lateral distribution of lipid components and other species of interest. However, quantitative analysis of FRET data with a topological model requires adequate choices [...] Read more.

Förster resonance energy transfer (FRET) is a powerful tool used for many problems in membrane biophysics, including characterization of the lateral distribution of lipid components and other species of interest. However, quantitative analysis of FRET data with a topological model requires adequate choices for the values of several input parameters, some of which are difficult to obtain experimentally in an independent manner. For this purpose, atomistic molecular dynamics (MD) simulations can be potentially useful as they provide direct detailed information on transverse probe localization, relative probe orientation, and membrane surface area, all of which are required for analysis of FRET data. This is illustrated here for the FRET pairs involving 1,6-diphenylhexatriene (DPH) as donor and either 1-palmitoyl,2-(6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] hexanoyl)- sn-glycero-3-phosphocholine (C6-NBD-PC) or 1-palmitoyl,2-(12-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]dodecanoyl)-sn-glycero-3-phosphocholine (C12-NBD-PC) as acceptors, in fluid vesicles of 1,2-dipalmitoyl-sn-3-glycerophosphocholine (DPPC, 50 °C). Incorporation of results from MD simulations improves the statistical quality of model fitting to the experimental FRET data. Furthermore, the decay of DPH in the presence of moderate amounts of C12-NBD-PC (>0.4 mol%) is consistent with non-random lateral distribution of the latter, at variance with C6-NBD-PC, for which aggregation is ruled out up to 2.5 mol% concentration. These conclusions are supported by analysis of NBD-PC fluorescence self-quenching. Implications regarding the relative utility of these probes in membrane studies are discussed. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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14 pages, 1133 KiB

Open AccessArticle

Chronic Supplementation of Paeonol Combined with Danshensu for the Improvement of Vascular Reactivity in the Cerebral Basilar Artery of Diabetic Rats

by Jing Hu^1,2,†, Ya-Ling Li^3,†, Zi-Lin Li^2,†, Hua Li¹, Xuan-Xuan Zhou¹, Peng-Cheng Qiu¹, Qian Yang¹ and Si-Wang Wang^1,*

¹ Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China

² General Hospital of Lanzhou Command, PLA, Lanzhou 730050, China

³ Department of Special Diagnosis, The Second Authority Clinic of Lanzhou Command, PLA, Lanzhou 730000, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14565-14578; https://doi.org/10.3390/ijms131114565 - 8 Nov 2012

Cited by 29 | Viewed by 6318

Abstract

One of the leading causes of death in the world is cerebrovascular disease. Numerous Chinese traditional medicines, such as Cortex Moutan (root bark of Paeonia suffruticosa Andrew) and Radix Salviae miltiorrhizae (root and rhizome of Salvia miltiorrhiza Bunge), protect against cerebrovascular diseases and [...] Read more.

One of the leading causes of death in the world is cerebrovascular disease. Numerous Chinese traditional medicines, such as Cortex Moutan (root bark of Paeonia suffruticosa Andrew) and Radix Salviae miltiorrhizae (root and rhizome of Salvia miltiorrhiza Bunge), protect against cerebrovascular diseases and exhibit anti-atherosclerotic effects. Traditional medicines have been routinely used for a long time in China. In addition, these two herbs are prescribed together in clinical practice. Therefore, the pharmacodynamic interactions between the active constituents of these two herbs, which are paeonol (Pae) and danshensu (DSS), should be particularly studied. The study of Pae and DSS can provide substantial foundations in understanding their mechanisms and empirical evidence to support clinical practice. This study investigated the effects and possible mechanisms of the pharmacodynamic interaction between Pae and DSS on cerebrovascular malfunctioning in diabetes. Experimental diabetes was induced in rats, which was then treated with Pae, DSS, and Pae + DSS for eight weeks. Afterward, cerebral arteries from all groups were isolated and equilibrated in an organ bath with Krebs buffer and ring tension. Effects of Pae, DSS, and Pae + DSS were observed on vessel relaxation with or without endothelium as well as on the basal tonus of vessels from normal and diabetic rats. Indexes about oxidative stress were also determined. We report that the cerebral arteries from diabetic rats show decreased vascular reactivity to acetylcholine (ACh) which was corrected in Pae, DSS, and Pae + DSS treated groups. Furthermore, phenylephrine (PE)-induced contraction response decreased in the treated groups. Phenylephrine and CaCl₂-induced vasoconstrictions are partially inhibited in the three treated groups under Ca²⁺-free medium. Pre-incubated with tetraethylammonium, a non-selective K⁺ channel blocker, the antagonized relaxation responses increased in DSS and Pae + DSS treated diabetic groups compared with those in diabetic and Pae-treated diabetic groups. In addition, superoxide dismutase activity and thiobarbituric acid reactive substances content significantly changed in the presence of Pae + DSS. We therefore conclude that both Pae and DSS treatments prevent diabetes-induced vascular damage. Furthermore, Pae + DSS prove to be the most efficient treatment regimen. The combination of Pae and DSS produce significant protective effects through the reduction of oxidative stress and through intracellular Ca²⁺ regulatory mechanisms. Full article

(This article belongs to the Section Biochemistry)

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27 pages, 2177 KiB

Open AccessReview

Glatiramer Acetate in Treatment of Multiple Sclerosis: A Toolbox of Random Co-Polymers for Targeting Inflammatory Mechanisms of both the Innate and Adaptive Immune System?

by Babak Jalilian, Halldór Bjarki Einarsson and Thomas Vorup-Jensen^*

Department of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, Building 1242, DK-8000, Aarhus C, Denmark

Int. J. Mol. Sci. 2012, 13(11), 14579-14605; https://doi.org/10.3390/ijms131114579 - 9 Nov 2012

Cited by 27 | Viewed by 16149

Abstract

Multiple sclerosis is a disease of the central nervous system, resulting in the demyelination of neurons, causing mild to severe symptoms. Several anti-inflammatory treatments now play a significant role in ameliorating the disease. Glatiramer acetate (GA) is a formulation of random polypeptide copolymers [...] Read more.

Multiple sclerosis is a disease of the central nervous system, resulting in the demyelination of neurons, causing mild to severe symptoms. Several anti-inflammatory treatments now play a significant role in ameliorating the disease. Glatiramer acetate (GA) is a formulation of random polypeptide copolymers for the treatment of relapsing-remitting MS by limiting the frequency of attacks. While evidence suggests the influence of GA on inflammatory responses, the targeted molecular mechanisms remain poorly understood. Here, we review the multiple pharmacological modes-of-actions of glatiramer acetate in treatment of multiple sclerosis. We discuss in particular a newly discovered interaction between the leukocyte-expressed integrin α_Mβ₂ (also called Mac-1, complement receptor 3, or CD11b/CD18) and perspectives on the GA co-polymers as an influence on the function of the innate immune system. Full article

(This article belongs to the Special Issue Recent Advances in the Research of Multiple Sclerosis)

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Graphical abstract

17 pages, 307 KiB

Open AccessReview

Roles of Nitric Oxide and Asymmetric Dimethylarginine in Pregnancy and Fetal Programming

by Li-Tung Huang^1,2,†, Chih-Sung Hsieh^3,4,†, Kow-Aung Chang⁵ and You-Lin Tain^1,6,*

¹ Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 800, Taiwan

² Department of Traditional Chinese Medicine, Chang Gung University, Linkow 333, Taiwan

³ Department of Pediatric Surgery, Pingtung Christian Hospital, Pingtung 900, Taiwan

⁴ Department of Nursing, MeiHo University, Pingtung 900, Taiwan

⁵ Department of Anesthesiology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 800, Taiwan

⁶ Center for Translational Research in Biomedical Sciences, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 800, Taiwan

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14606-14622; https://doi.org/10.3390/ijms131114606 - 9 Nov 2012

Cited by 64 | Viewed by 9196

Abstract

Nitric oxide (NO) regulates placental blood flow and actively participates in trophoblast invasion and placental development. Asymmetric dimethylarginine (ADMA) can inhibit NO synthase, which generates NO. ADMA has been associated with uterine artery flow disturbances such as preeclampsia. Substantial experimental evidence has reliably [...] Read more.

Nitric oxide (NO) regulates placental blood flow and actively participates in trophoblast invasion and placental development. Asymmetric dimethylarginine (ADMA) can inhibit NO synthase, which generates NO. ADMA has been associated with uterine artery flow disturbances such as preeclampsia. Substantial experimental evidence has reliably supported the hypothesis that an adverse in utero environment plays a role in postnatal physiological and pathophysiological programming. Growing evidence suggests that the placental nitrergic system is involved in epigenetic fetal programming. In this review, we discuss the roles of NO and ADMA in normal and compromised pregnancies as well as the link between placental insufficiency and epigenetic fetal programming. Full article

(This article belongs to the Special Issue ADMA and Nitrergic System)

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11 pages, 1185 KiB

Open AccessArticle

Molecular Systematics of Genus Atractylodes (Compositae, Cardueae): Evidence from Internal Transcribed Spacer (ITS) and trnL-F Sequences

by Hua-Sheng Peng^1,2, Qing-Jun Yuan¹, Qian-Quan Li¹ and Lu-Qi Huang^1,*

¹ Institute of Chinese Materia Medica, China Academy of Chinese Medical Science, Beijing 100700, China

² Department of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230031, China

Int. J. Mol. Sci. 2012, 13(11), 14623-14633; https://doi.org/10.3390/ijms131114623 - 9 Nov 2012

Cited by 27 | Viewed by 6208

Abstract

To determine the evolutionary relationships among all members of the genus Atractylodes (Compositae, Cardueae), we conducted molecular phylogenetic analyses of one nuclear DNA (nrDNA) region (internal transcribed spacer, ITS) and one chloroplast DNA (cpDNA) region (intergenic spacer region of trnL-F [...] Read more.

To determine the evolutionary relationships among all members of the genus Atractylodes (Compositae, Cardueae), we conducted molecular phylogenetic analyses of one nuclear DNA (nrDNA) region (internal transcribed spacer, ITS) and one chloroplast DNA (cpDNA) region (intergenic spacer region of trnL-F). In ITS and ITS + trnL-F trees, all members of Atractylodes form a monophyletic clade. Atractylodes is a sister group of the Carlina and Atractylis branch. Atractylodes species were distributed among three clades: (1) A. carlinoides (located in the lowest base of the Atractylodes phylogenetic tree), (2) A. macrocephala, and (3) the A. lancea complex, including A. japonica, A. coreana, A. lancea, A. lancea subsp. luotianensis, and A. chinensis. The taxonomic controversy over the classification of species of Atractylodes is mainly concentrated in the A. lancea complex. In base on molecular results, the intraspecific division of Atractylodes lancea is not supported, and A. coreana should be treated as a synonym A. chinensis. Full article

(This article belongs to the Section Biochemistry)

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14 pages, 899 KiB

Open AccessArticle

Radix Paeoniae Rubra and Radix Paeoniae Alba Attenuate CCl₄-Induced Acute Liver Injury: An Ultra-Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS) Based Metabolomic Approach for the Pharmacodynamic Study of Traditional Chinese Medicines (TCMs)

by Rui Wang¹, Ai-Zhen Xiong^2,3, Zhong-Qiu Teng⁴, Qi-Wei Yang¹, Yan-Hong Shi¹ and Li Yang^2,3,*

¹ School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

² The Ministry of Education (MOE) Key Laboratory for Standardization of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

³ The State Administration of Traditional Chinese Medicine (SATCM), Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

⁴ School of Life Science, Beijing Institute of Technology, Beijing 100081, China

Int. J. Mol. Sci. 2012, 13(11), 14634-14647; https://doi.org/10.3390/ijms131114634 - 9 Nov 2012

Cited by 57 | Viewed by 9230

Abstract

Metabolomics has been frequently used in pharmacodynamic studies, especially those on traditional Chinese medicine (TCM). Radix Paeoniae Alba and Radix Paeoniae Rubra are popularly used in TCM, and both have hepatoprotective effects. In this study, a CCl₄-induced acute liver injury rat [...] Read more.

Metabolomics has been frequently used in pharmacodynamic studies, especially those on traditional Chinese medicine (TCM). Radix Paeoniae Alba and Radix Paeoniae Rubra are popularly used in TCM, and both have hepatoprotective effects. In this study, a CCl₄-induced acute liver injury rat model was established and confirmed by the observed serum aminotransferase activities. The metabolomics approach was applied to study the influence of Radix Paeoniae Alba and Radix Paeoniae Rubra on the metabolic changes in rats with acute liver injury. The partial least-squares-discriminant analysis (PLS-DA) of rat serum and their ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) fingerprints allowed discrimination of controlled, acute liver injury-model rats after administration of the two types of TCMs. The time-dependent PLS-DA plots showed that the changes in the metabolic patterns of the rats, which were administered with the TCMs, had stabilized within 2 h after they received the intraperitoneal CCl₄ injection. The results indicated the protective effect of TCMs against liver injury. Several potential biomarkers were detected and identified, which included creatine, deoxycholic acid, choline, 5-methylenetetrahydrofolate, folic acid, and glycocholic acid. The physiological significance of these metabolic changes was discussed. Full article

(This article belongs to the Section Biochemistry)

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19 pages, 327 KiB

Open AccessReview

An Advanced Electron Spin Resonance (ESR) Spin-Trapping and LC/(ESR)/MS Technique for the Study of Lipid Peroxidation

by Yi Xu, Yan Gu and Steven Y. Qian^*

Department of Pharmaceutical Sciences, College of Pharmacy, Nursing and Allied Sciences, North Dakota State University, Fargo, ND 58108, USA

Int. J. Mol. Sci. 2012, 13(11), 14648-14666; https://doi.org/10.3390/ijms131114648 - 12 Nov 2012

Cited by 22 | Viewed by 8717

Abstract

There are two types of nutritionally important polyunsaturated fatty acids (PUFAs), namely ω-6s and ω-3s. PUFAs and their metabolites generated from lipid peroxidation via cyclooxygenase (COX) and lipoxygenase (LOX) are believed to be involved in a variety of physiological and pathological processes in [...] Read more.

There are two types of nutritionally important polyunsaturated fatty acids (PUFAs), namely ω-6s and ω-3s. PUFAs and their metabolites generated from lipid peroxidation via cyclooxygenase (COX) and lipoxygenase (LOX) are believed to be involved in a variety of physiological and pathological processes in the human body. Both COX- and LOX-catalyzed PUFA peroxidation are complex events that generate a series of radicals, which may then bind proteins, target DNA/RNA, and lead to a number of biological changes. However, due to the lack of an appropriate method, it was not possible until recently to identify the short-lived PUFA-derived radicals in COX-/LOX-catalyzed peroxidation. Failure to characterize free radicals during peroxidation has greatly restricted our knowledge about COX/LOX biology in human health. Here we review the development and refinement of combined ESR spin trapping and LC/ESR/MS to characterize PUFA-derived radicals formed from in vitro (cell-free) peroxidation. We also present the most recent approach for studying peroxidation in cells which allows us to directly assess the potential bioactivity of PUFA-derived free radicals. This advanced technique has resulted in a major breakthrough in radical structural characterization, as well as assessment of free radical-associated cell growth response, thereby greatly improving our knowledge of PUFAs, COX-/LOX-catalyzed lipid peroxidation, and their related biological consequences. Full article

(This article belongs to the Special Issue Advances in Free Radicals in Biology and Medicine)

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12 pages, 1155 KiB

Open AccessArticle

Role of Btg2 in the Progression of a PDGF-Induced Oligodendroglioma Model

by Irene Appolloni¹, Sebastiano Curreli², Sara Caviglia², Manuela Barilari¹, Eleonora Gambini², Aldo Pagano^1,2 and Paolo Malatesta^1,2,*

¹ San Martino-IST, Largo Rosanna Benzi 10, 16132 Genoa, Italy

² Department of Experimental Medicine (DiMES), University of Genoa, 16132 Genoa, Italy

Int. J. Mol. Sci. 2012, 13(11), 14667-14678; https://doi.org/10.3390/ijms131114667 - 12 Nov 2012

Cited by 8 | Viewed by 6748

Abstract

Tumor progression is a key aspect in oncology. Not even the overexpression of a powerful oncogenic stimulus such as platelet derived growth factor-B (PDGF-B) is sufficient per se to confer full malignancy to cells. In previous studies we showed that neural progenitors overexpressing [...] Read more.

Tumor progression is a key aspect in oncology. Not even the overexpression of a powerful oncogenic stimulus such as platelet derived growth factor-B (PDGF-B) is sufficient per se to confer full malignancy to cells. In previous studies we showed that neural progenitors overexpressing PDGF-B need to undergo progression to acquire the capability to give rise to secondary tumor following transplant. By comparing the expression profile of PDGF-expressing cells before and after progression, we found that progressed tumors consistently downregulate the expression of the antiproliferative gene Btg2. We therefore tested whether the downregulation of Btg2 is sufficient and necessary for glioma progression with loss and gain of function experiments. Our results show that downregulation of Btg2 is not sufficient but is necessary for tumor progression since the re-introduction of Btg2 in fully progressed tumors dramatically impairs their gliomagenic potential. These results suggest an important role of Btg2 in glioma progression. Accordingly with this view, the analysis of public datasets of human gliomas showed that reduced level of Btg2 expression correlates with a significantly worse prognosis. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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19 pages, 328 KiB

Open AccessArticle

Dual Bioactivities of Essential Oil Extracted from the Leaves of Artemisia argyi as an Antimelanogenic versus Antioxidant Agent and Chemical Composition Analysis by GC/MS

by Huey-Chun Huang¹, Hsiao-Fen Wang², Kuang-Hway Yih³, Long-Zen Chang^4,* and Tsong-Min Chang^3,*

¹ Department of Medical Laboratory Science and Biotechnology, China Medical University, No 91 Hsueh-Shih Road, Taichung 40402, Taiwan

² Department of Hair styling & Design, Hung Kuang University, No. 34, Chung-Chie Road, Shalu, Taichung 43302, Taiwan

³ Department of Applied Cosmetology & Master Program of Cosmetic Science, Hung Kuang University, No. 34, Chung-Chie Road, Shalu, Taichung 43302, Taiwan

⁴ General of Agriculture Bureau of Taichung City, No. 89, Sec 2, Taichung Port Road, Xitun Dist., Taichung 40701, Taiwan

Int. J. Mol. Sci. 2012, 13(11), 14679-14697; https://doi.org/10.3390/ijms131114679 - 12 Nov 2012

Cited by 92 | Viewed by 10730

Abstract

The study was aimed at investigating the antimelanogenic and antioxidant properties of essential oil when extracted from the leaves of Artemisia argyi, then analyzing the chemical composition of the essential oil. The inhibitory effect of the essential oil on melanogenesis was evaluated [...] Read more.

The study was aimed at investigating the antimelanogenic and antioxidant properties of essential oil when extracted from the leaves of Artemisia argyi, then analyzing the chemical composition of the essential oil. The inhibitory effect of the essential oil on melanogenesis was evaluated by a mushroom tyrosinase activity assay and B16F10 melanoma cell model. The antioxidant capacity of the essential oil was assayed by spectrophotometric analysis, and the volatile chemical composition of the essential oil was analyzed with gas chromatography-mass spectrometry (GC/MS). The results revealed that the essential oil significantly inhibits mushroom tyrosinase activity (IC₅₀ = 19.16 mg/mL), down-regulates B16F10 intracellular tyrosinase activity and decreases the amount of melanin content in a dose-dependent pattern. Furthermore, the essential oil significantly scavenged 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) ABTS radicals, showed an apparent reduction power as compared with metal-ion chelating activities. The chemicals constituents in the essential oil are ether (23.66%), alcohols (16.72%), sesquiterpenes (15.21%), esters (11.78%), monoterpenes (11.63%), ketones (6.09%), aromatic compounds (5.01%), and account for a 90.10% analysis of its chemical composition. It is predicted that eucalyptol and the other constituents, except for alcohols, in the essential oil may contribute to its antioxidant activities. The results indicated that essential oil extracted from A. argyi leaves decreased melanin production in B16F10 cells and showed potent antioxidant activity. The essential oil can thereby be applied as an inhibitor of melanogenesis and could also act as a natural antioxidant in skin care products. Full article

(This article belongs to the Section Biochemistry)

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9 pages, 214 KiB

Open AccessArticle

Non-Alcoholic Fatty Liver Disease Is not Related to the Incidence of Diabetic Nephropathy in Type 2 Diabetes

by Yu-Tao Zhan^1,†, Chuan Zhang^2,†, Li Li¹, Chun-Shan Bi¹, Xin Song³ and Shu-Tian Zhang^4,*

¹ Department of Gastroenterology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China

² Department of Gastroenterology, Beijing Chaoyang Hospital Jingxi Campus, Capital Medical University, Beijing 100430, China

³ Department of Clinic Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China

⁴ Department of Gastroenterology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14698-14706; https://doi.org/10.3390/ijms131114698 - 12 Nov 2012

Cited by 19 | Viewed by 6166

Abstract

To analyze the association between non-alcoholic fatty liver disease (NAFLD) and the incidence of diabetic nephropathy in patients with type 2 diabetes, the incidence of diabetic nephropathy was assessed in 413 type 2 diabetic patients, by testing the 24 h urinary albumin excretion [...] Read more.

To analyze the association between non-alcoholic fatty liver disease (NAFLD) and the incidence of diabetic nephropathy in patients with type 2 diabetes, the incidence of diabetic nephropathy was assessed in 413 type 2 diabetic patients, by testing the 24 h urinary albumin excretion rate (UAER). The NAFLD was diagnosed based on patient’s medical history and liver ultrasound. The difference in diabetic nephropathy incidence between patients with and without NAFLD was tested by χ². Multivariate logistic regression analysis was used to assess the factors associated with diabetic nephropathy among type 2 diabetic patients. Total 363 out of 413 type 2 diabetic patients were enrolled in this study. The incidences of NAFLD and diabetic nephropathy in participants were approximately 56% (202/363) and 38% (137/363) respectively, and there was no significant difference in the prevalence of diabetic nephropathy between patients with and without NAFLD (37.1% vs. 38.5%, p = 0.787). The duration of diabetes (odds ratio [OR] 1.065, 95% confidence interval [CI] 1.014–1.120, p = 0.012), waist circumference (OR 1.077, 95% CI 1.040–1.116, p = 0.000), and fasting blood glucose (FBG; OR 1.136, 95% CI 1.023–1.1262, p = 0.017) were significantly associated with diabetic nephropathy, whereas sex, high blood pressure, total cholesterol (TC), triglyceride (TG), and ankle brachial pressure index (ABI) were not significantly associated with the disorder. The present results suggest that NAFLD is not related to the incidence of diabetic nephropathy in type 2 diabetes, but the duration of diabetes, waist circumference, and FBG are important factors for diabetic nephropathy in type 2 diabetes. Full article

(This article belongs to the Section Biochemistry)

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16 pages, 233 KiB

Open AccessArticle

Biological Activities of Toninia candida and Usnea barbata Together with Their Norstictic Acid and Usnic Acid Constituents

by Branislav Ranković¹, Marijana Kosanić^1,*, Tatjana Stanojković², Perica Vasiljević³ and Nedeljko Manojlović⁴

¹ Department of Biology, Faculty of Science, University of Kragujevac, 34000 Kragujevac, Serbia

² Institute of Oncology and Radiology of Serbia, 11000 Belgrade, Serbia

³ Department of Biology and Ecology, Faculty of Sciences and Mathematics, University of Niš, 18000 Niš, Serbia

⁴ Department of Pharmacy, Faculty of Medical Sciences, University of Kragujevac, 34000 Kragujevac, Serbia

Int. J. Mol. Sci. 2012, 13(11), 14707-14722; https://doi.org/10.3390/ijms131114707 - 12 Nov 2012

Cited by 110 | Viewed by 10081

Abstract

The aim of this study was to investigate the chemical composition of acetone extracts of the lichens Toninia candida and Usnea barbata and in vitro antioxidant, antimicrobial, and anticancer activities of these extracts together with some of their major metabolites. The chemical composition [...] Read more.

The aim of this study was to investigate the chemical composition of acetone extracts of the lichens Toninia candida and Usnea barbata and in vitro antioxidant, antimicrobial, and anticancer activities of these extracts together with some of their major metabolites. The chemical composition of T. candida and U. barbata extracts was determined using HPLC-UV analysis. The major phenolic compounds in these extracts were norstictic acid (T. candida) and usnic acid (U. barbata). Antioxidant activity was evaluated by free radical scavenging, superoxide anion radical scavenging, reducing power and determination of total phenolic compounds. Results of the study proved that norstictic acid had the largest antioxidant activity. The total content of phenols in the extracts was determined as the pyrocatechol equivalent. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration using the broth microdilution method. The most active was usnic acid with minimum inhibitory concentration values ranging from 0.0008 to 0.5 mg/mL. Anticancer activity was tested against FemX (human melanoma) and LS174 (human colon carcinoma) cell lines using the microculture tetrazolium test. Usnic acid was found to have the strongest anticancer activity towards both cell lines with IC₅₀ values of 12.72 and 15.66 μg/mL. Full article

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19 pages, 718 KiB

Open AccessArticle

Influence of Dose on Particle Size and Optical Properties of Colloidal Platinum Nanoparticles

by Elham Gharibshahi and Elias Saion^*

Department of Physics, University Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 14723-14741; https://doi.org/10.3390/ijms131114723 - 12 Nov 2012

Cited by 134 | Viewed by 14036

Abstract

Attempts to produce colloidal platinum nanoparticles by using steady absorption spectra with various chemical-based reduction methods often resulted in the fast disappearance of the absorption maxima leaving reduced platinum nanoparticles with little information on their optical properties. We synthesized colloidal platinum nanoparticles in [...] Read more.

Attempts to produce colloidal platinum nanoparticles by using steady absorption spectra with various chemical-based reduction methods often resulted in the fast disappearance of the absorption maxima leaving reduced platinum nanoparticles with little information on their optical properties. We synthesized colloidal platinum nanoparticles in an aqueous solution of polyvinyl pyrrolidone by gamma radiolytic reduction method, which produced steady absorption spectra of fully reduced and highly pure platinum nanoparticles free from by-product impurities or reducing agent contamination. The average particle size was found to be in the range of 3.4–5.3 nm and decreased with increasing dose due to the domination of nucleation over ion association in the formation of metal nanoparticles by the gamma radiolytic reduction method. The platinum nanoparticles exhibit optical absorption spectra with two absorption peaks centered at about 216 and 264 nm and the peaks blue shifted to lower wavelengths with decreasing particle size. The absorption spectra of platinum nanoparticles were also calculated using quantum mechanical treatment and coincidently a good agreement was obtained between the calculated and measured absorption peaks at various particle sizes. This indicates that the 216 and 264-nm absorption peaks of platinum nanoparticles conceivably originated from the intra-band transitions of conduction electrons of (n = 5, l = 2) and (n = 6, l = 0) energy states respectively to higher energy states. The absorption energies, i.e., conduction band energies of platinum nanoparticles derived from the absorption peaks increased with increasing dose and decreased with increasing particle size. Full article

(This article belongs to the Section Materials Science)

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24 pages, 1337 KiB

Open AccessReview

Single Molecule Fluorescence Detection and Tracking in Mammalian Cells: The State-of-the-Art and Future Perspectives

by Marisa L. Martin-Fernandez^* and David T. Clarke

Central Laser Facility, Science & Technology Facilities Council, Research Complex at Harwell, Rutherford Appleton Laboratory, Harwell Oxford, Didcot OX11 0QX, UK

Int. J. Mol. Sci. 2012, 13(11), 14742-14765; https://doi.org/10.3390/ijms131114742 - 13 Nov 2012

Cited by 25 | Viewed by 9483

Abstract

Insights from single-molecule tracking in mammalian cells have the potential to greatly contribute to our understanding of the dynamic behavior of many protein families and networks which are key therapeutic targets of the pharmaceutical industry. This is particularly so at the plasma membrane, [...] Read more.

Insights from single-molecule tracking in mammalian cells have the potential to greatly contribute to our understanding of the dynamic behavior of many protein families and networks which are key therapeutic targets of the pharmaceutical industry. This is particularly so at the plasma membrane, where the method has begun to elucidate the mechanisms governing the molecular interactions that underpin many fundamental processes within the cell, including signal transduction, receptor recognition, cell-cell adhesion, etc. However, despite much progress, single-molecule tracking faces challenges in mammalian samples that hinder its general application in the biomedical sciences. Much work has recently focused on improving the methods for fluorescent tagging of target molecules, detection and localization of tagged molecules, which appear as diffraction-limited spots in charge-coupled device (CCD) images, and objectively establishing the correspondence between moving particles in a sequence of image frames to follow their diffusive behavior. In this review we outline the state-of-the-art in the field and discuss the advantages and limitations of the methods available in the context of specific applications, aiming at helping researchers unfamiliar with single molecules methods to plan out their experiments. Full article

(This article belongs to the Special Issue Advances in Single Molecule Spectroscopy)

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Graphical abstract

22 pages, 401 KiB

Open AccessArticle

Alzheimer’s Disease-Associated Neurotoxic Peptide Amyloid-Β Impairs Base Excision Repair in Human Neuroblastoma Cells

by Anne Forestier¹, Thierry Douki¹, Sylvie Sauvaigo¹, Viviana De Rosa¹, Christine Demeilliers² and Walid Rachidi^1,*

¹ Nucleic Acids Lesions Laboratory, SCIB/INAC, CEA, Joseph Fourier University-Grenoble 1, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France

² INSERM U1055, Joseph Fourier University-Grenoble 1, 38000 Grenoble, France

Int. J. Mol. Sci. 2012, 13(11), 14766-14787; https://doi.org/10.3390/ijms131114766 - 13 Nov 2012

Cited by 23 | Viewed by 7824

Abstract

Alzheimer’s disease (AD) is the leading cause of dementia in developed countries. It is characterized by two major pathological hallmarks, one of which is the extracellular aggregation of the neurotoxic peptide amyloid-β (Aβ), which is known to generate oxidative stress. In this study, [...] Read more.

Alzheimer’s disease (AD) is the leading cause of dementia in developed countries. It is characterized by two major pathological hallmarks, one of which is the extracellular aggregation of the neurotoxic peptide amyloid-β (Aβ), which is known to generate oxidative stress. In this study, we showed that the presence of Aβ in a neuroblastoma cell line led to an increase in both nuclear and mitochondrial DNA damage. Unexpectedly, a concomitant decrease in basal level of base excision repair, a major route for repairing oxidative DNA damage, was observed at the levels of both gene expression and protein activity. Moreover, the addition of copper sulfate or hydrogen peroxide, used to mimic the oxidative stress observed in AD-affected brains, potentiates Aβ-mediated perturbation of DNA damage/repair systems in the “Aβ cell line”. Taken together, these findings indicate that Aβ could act as double-edged sword by both increasing oxidative nuclear/mitochondrial damage and preventing its repair. The synergistic effects of increased ROS production, accumulated DNA damage and impaired DNA repair could participate in, and partly explain, the massive loss of neurons observed in Alzheimer’s disease since both oxidative stress and DNA damage can trigger apoptosis. Full article

(This article belongs to the Special Issue DNA Damage and Repair in Degenerative Diseases)

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25 pages, 676 KiB

Open AccessArticle

Phosphatidylethanol in Blood as a Marker of Chronic Alcohol Use: A Systematic Review and Meta-Analysis

by Guido Viel^*,†

, Rafael Boscolo-Berto^*,†, Giovanni Cecchetto, Paolo Fais, Alessandro Nalesso and Santo Davide Ferrara

¹ Department of Molecular Medicine, Forensic Toxicology and Antidoping Unit, University of Padova, Via Falloppio 50, Padova 35121, Italy

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14788-14812; https://doi.org/10.3390/ijms131114788 - 13 Nov 2012

Cited by 210 | Viewed by 17882

Abstract

The present paper aims at a systematic review of the current knowledge on phosphatidylethanol (PEth) in blood as a direct marker of chronic alcohol use and abuse. In March 2012, the search through “MeSH” and “free-text” protocols in the databases Medline/PubMed, SCOPUS, Web [...] Read more.

The present paper aims at a systematic review of the current knowledge on phosphatidylethanol (PEth) in blood as a direct marker of chronic alcohol use and abuse. In March 2012, the search through “MeSH” and “free-text” protocols in the databases Medline/PubMed, SCOPUS, Web of Science, and Ovid/Embase, combining the terms phosphatidylethanol and alcohol, provided 444 records, 58 of which fulfilled the inclusion criteria and were used to summarize the current evidence on the formation, distribution and degradation of PEth in human blood: (1), the presence and distribution of different PEth molecular species (2), the most diffused analytical methods devoted to PEth identification and quantization (3), the clinical efficiency of total PEth quantification as a marker of chronic excessive drinking (4), and the potential utility of this marker for identifying binge drinking behaviors (5). Twelve papers were included in the meta-analysis and the mean (M) and 95% confidence interval (CI) of total PEth concentrations in social drinkers (DAI ≤ 60 g/die; M = 0.288 µM; CI 0.208–0.367 µM) and heavy drinkers (DAI > 60 g/die; M = 3.897 µM; CI 2.404–5.391 µM) were calculated. The present analysis demonstrates a good clinical efficiency of PEth for detecting chronic heavy drinking. Full article

(This article belongs to the Special Issue Phospholipids: Molecular Sciences 2012)

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15 pages, 323 KiB

Open AccessArticle

The Murine PSE/TATA-Dependent Transcriptome: Evidence of Functional Homologies with Its Human Counterpart

by Maria Jessica Bruzzone¹, Paola Gavazzo², Sara Massone¹, Carolina Balbi¹, Federico Villa¹, Anastasia Conti³, Giorgio Dieci³

, Ranieri Cancedda^1,4 and Aldo Pagano^1,4,*

¹ Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy

² Institute of Biophysics, National Council of Research (CNR), Via De Marini 6, 16149 Genoa, Italy

³ Department of Biosciences, University of Parma, Parco Area delle Scienze 23/A, 43124 Parma, Italy

⁴ San Martino-IST, Largo Rosanna Benzi 10, 16132 Genoa, Italy

Int. J. Mol. Sci. 2012, 13(11), 14813-14827; https://doi.org/10.3390/ijms131114813 - 13 Nov 2012

Cited by 2 | Viewed by 5726

Abstract

A series of recent studies demonstrated an unexpectedly high frequency of intronic RNA polymerase (pol) III transcription units spread throughout the human genome. The investigation of a subset of these transcripts revealed their tissue/cell-specific transcription together with the involvement in relevant physiopathological pathways. [...] Read more.

A series of recent studies demonstrated an unexpectedly high frequency of intronic RNA polymerase (pol) III transcription units spread throughout the human genome. The investigation of a subset of these transcripts revealed their tissue/cell-specific transcription together with the involvement in relevant physiopathological pathways. Despite this evidence, these transcripts did not seem to have murine orthologs, based on their nucleotide sequence, resulting in a limitation of the experimental approaches aimed to study their function. In this work, we have extended our investigation to the murine genome identifying 121 pairs of mouse/human transcripts displaying syntenic subchromosomal localization. The analysis in silico of this set of putative noncoding (nc)RNAs suggest their association with alternative splicing as suggested by recent experimental evidence. The investigation of one of these pairs taken as experimental model in mouse hippocampal neurons provided evidence of a human/mouse functional homology that does not depend on underlying sequence conservation. In this light, the collection of transcriptional units here reported can be considered as a novel source for the identification and the study of novel regulatory elements involved in relevant biological processes. Full article

(This article belongs to the Section Biochemistry)

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17 pages, 298 KiB

Open AccessArticle

Involvement of Salicylic Acid on Antioxidant and Anticancer Properties, Anthocyanin Production and Chalcone Synthase Activity in Ginger (Zingiber officinale Roscoe) Varieties

by Ali Ghasemzadeh^*

, Hawa Z. E. Jaafar^* and Ehsan Karimi

Department of Crop Science, Faculty of Agriculture, University Putra Malaysia, Serdang 43400, Selangor, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 14828-14844; https://doi.org/10.3390/ijms131114828 - 13 Nov 2012

Cited by 78 | Viewed by 10787

Abstract

The effect of foliar application of salicylic acid (SA) at different concentrations (10⁻³ M and 10⁻⁵ M) was investigated on the production of secondary metabolites (flavonoids), chalcone synthase (CHS) activity, antioxidant activity and anticancer activity (against breast cancer cell lines MCF-7 [...] Read more.

The effect of foliar application of salicylic acid (SA) at different concentrations (10⁻³ M and 10⁻⁵ M) was investigated on the production of secondary metabolites (flavonoids), chalcone synthase (CHS) activity, antioxidant activity and anticancer activity (against breast cancer cell lines MCF-7 and MDA-MB-231) in two varieties of Malaysian ginger, namely Halia Bentong and Halia Bara. The results of high performance liquid chromatography (HPLC) analysis showed that application of SA induced the synthesis of anthocyanin and fisetin in both varieties. Anthocyanin and fisetin were not detected in the control plants. Accordingly, the concentrations of some flavonoids (rutin and apigenin) decreased significantly in plants treated with different concentrations of SA. The present study showed that SA enhanced the chalcone synthase (CHS) enzyme activity (involving flavonoid synthesis) and recorded the highest activity value of 5.77 nkat /mg protein in Halia Bara with the 10⁻⁵ M SA treatment. As the SA concentration was decreased from 10⁻³ M to 10⁻⁵ M, the free radical scavenging power (FRAP) increased about 23% in Halia Bentong and 10.6% in Halia Bara. At a concentration of 350 μg mL⁻¹, the DPPH antioxidant activity recorded the highest value of 58.30%–72.90% with the 10⁻⁵ M SA treatment followed by the 10⁻³ M SA (52.14%–63.66%) treatment. The lowest value was recorded in the untreated control plants (42.5%–46.7%). These results indicate that SA can act not only as an inducer but also as an inhibitor of secondary metabolites. Meanwhile, the highest anticancer activity against MCF-7 and MDA-MB-231 cell lines was observed for H. Bara extracts treated with 10⁻⁵ M SA with values of 61.53 and 59.88%, respectively. The results suggest that the high anticancer activity in these varieties may be related to the high concentration of potent anticancer components including fisetin and anthocyanin. The results thus indicate that the synthesis of flavonoids in ginger can be increased by foliar application of SA in a controlled environment and that the anticancer activity in young ginger extracts could be improved. Full article

(This article belongs to the Section Biochemistry)

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20 pages, 309 KiB

Open AccessReview

The Role of Vascular Endothelial Growth Factor A Polymorphisms in Breast Cancer

by Doonyapat Sa-nguanraksa and Pornchai O-charoenrat^*

Division of Head-Neck and Breast Surgery, Department of Surgery, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand

Int. J. Mol. Sci. 2012, 13(11), 14845-14864; https://doi.org/10.3390/ijms131114845 - 13 Nov 2012

Cited by 43 | Viewed by 10303

Abstract

Breast cancer is the most common cancer in females and the leading cause of cancer death in women worldwide. Angiogenesis, the formation of new blood vessels, plays an important role in the development and progression of breast cancer. Vascular endothelial growth factor A [...] Read more.

Breast cancer is the most common cancer in females and the leading cause of cancer death in women worldwide. Angiogenesis, the formation of new blood vessels, plays an important role in the development and progression of breast cancer. Vascular endothelial growth factor A (VEGFA), the key modulator of angiogenesis, is highly expressed in cancer tissue and correlates with its more aggressive features. Polymorphisms of VEGFA alter the levels of expression and subsequently influence the susceptibility and aggressiveness of breast cancer. Assessment of VEGFA polymorphisms may be used for the identification of patients suitable for anti-VEGFA therapy. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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6 pages, 185 KiB

Open AccessArticle

New Triterpenoids with Cytotoxic Activity from Actinidia Valvata

by Li-Ping Qu^†, Guo-Yin Zheng^†, Yong-Hua Su, Hui-Qing Zhang, Yan-Long Yang, Hai-Liang Xin and Chang-Quan Ling^*

¹ Department of Traditional Chinese Medicine, Changhai Hospital of Second Military Medical University, Shanghai 200433, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14865-14870; https://doi.org/10.3390/ijms131114865 - 13 Nov 2012

Cited by 7 | Viewed by 5463

Abstract

Two new triterpenoids, 30-O-β-D-glucopyranosyloxy-2α,3α,24-trihydroxyurs-12, 18-diene-28-oic acid O-β-D-glucopyranosyl ester (1) and 2α,3β,3,30-tetrahydroxyurs-12, 18-diene-28-oic acid O-β-D-glucopyranosyl ester (2) were isolated from roots of Actinidia valvata Dunn. Their structures were elucidated by means of extensive spectroscopic studies. Both [...] Read more.

Two new triterpenoids, 30-O-β-D-glucopyranosyloxy-2α,3α,24-trihydroxyurs-12, 18-diene-28-oic acid O-β-D-glucopyranosyl ester (1) and 2α,3β,3,30-tetrahydroxyurs-12, 18-diene-28-oic acid O-β-D-glucopyranosyl ester (2) were isolated from roots of Actinidia valvata Dunn. Their structures were elucidated by means of extensive spectroscopic studies. Both these two new compounds showed moderate cytotoxic activity in vitro against BEL-7402 and SMMC-7721 tumor cell line. Full article

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18 pages, 475 KiB

Open AccessArticle

Influence of Chemical Extraction on Rheological Behavior, Viscoelastic Properties and Functional Characteristics of Natural Heteropolysaccharide/Protein Polymer from Durio zibethinus Seed

by Bahareh Tabatabaee Amid and Hamed Mirhosseini^*

Department of Food Technology, Faculty of Food Science and Technology, University Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 14871-14888; https://doi.org/10.3390/ijms131114871 - 13 Nov 2012

Cited by 17 | Viewed by 5811

Abstract

In recent years, the demand for a natural plant-based polymer with potential functions from plant sources has increased considerably. The main objective of the current study was to study the effect of chemical extraction conditions on the rheological and functional properties of the [...] Read more.

In recent years, the demand for a natural plant-based polymer with potential functions from plant sources has increased considerably. The main objective of the current study was to study the effect of chemical extraction conditions on the rheological and functional properties of the heteropolysaccharide/protein biopolymer from durian (Durio zibethinus) seed. The efficiency of different extraction conditions was determined by assessing the extraction yield, protein content, solubility, rheological properties and viscoelastic behavior of the natural polymer from durian seed. The present study revealed that the soaking process had a more significant (p < 0.05) effect than the decolorizing process on the rheological and functional properties of the natural polymer. The considerable changes in the rheological and functional properties of the natural polymer could be due to the significant (p < 0.05) effect of the chemical extraction variables on the protein fraction present in the molecular structure of the natural polymer from durian seed. The natural polymer from durian seed had a more elastic (or gel like) behavior compared to the viscous (liquid like) behavior at low frequency. The present study revealed that the natural heteropolysaccharide/protein polymer from durian seed had a relatively low solubility ranging from 9.1% to 36.0%. This might be due to the presence of impurities, insoluble matter and large particles present in the chemical structure of the natural polymer from durian seed. Full article

(This article belongs to the Section Biochemistry)

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9 pages, 481 KiB

Open AccessArticle

Optimization of Lipase Production by Burkholderia sp. Using Response Surface Methodology

by Chia-Feng Lo, Chi-Yang Yu, I-Ching Kuan and Shiow-Ling Lee^*

Department of Bioengineering, Tatung University, No 40, Sec. 3, Zhongshan N. Rd., Taipei 104, Taiwan

Int. J. Mol. Sci. 2012, 13(11), 14889-14897; https://doi.org/10.3390/ijms131114889 - 13 Nov 2012

Cited by 42 | Viewed by 7729

Abstract

Response surface methodology (RSM) was employed to optimize the extracellular lipase production by Burkholderia sp. HL-10. Preliminary tests showed that olive oil, tryptone and Tween-80 exhibited significant effects on the lipase production. The optimum concentrations of these three components were determined using a [...] Read more.

Response surface methodology (RSM) was employed to optimize the extracellular lipase production by Burkholderia sp. HL-10. Preliminary tests showed that olive oil, tryptone and Tween-80 exhibited significant effects on the lipase production. The optimum concentrations of these three components were determined using a faced-centered central composite design (FCCCD). The analysis of variance revealed that the established model was significant (p < 0.01). The optimized medium containing 0.65% olive oil (v/v), 2.42% tryptone (w/v) and 0.15% Tween-80 (v/v) resulted in a maximum activity of 122.3 U/mL, about three fold higher than that in basal medium. Approximately 99% of validity of the predicted value was achieved. Full article

(This article belongs to the Special Issue Enzyme Optimization and Immobilization)

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19 pages, 829 KiB

Open AccessReview

A DNA Repair BRCA1 Estrogen Receptor and Targeted Therapy in Breast Cancer

by Adisorn Ratanaphan

Laboratory of Pharmaceutical Biotechnology, Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand

Int. J. Mol. Sci. 2012, 13(11), 14898-14916; https://doi.org/10.3390/ijms131114898 - 14 Nov 2012

Cited by 22 | Viewed by 14086

Abstract

BRCA1 is a key mediator of DNA repair pathways and participates in the maintenance of the genomic integrity of cells. The control of DNA damage repair mechanisms by BRCA1 is of great interest since molecular defects in this pathway may reflect a predictive [...] Read more.

BRCA1 is a key mediator of DNA repair pathways and participates in the maintenance of the genomic integrity of cells. The control of DNA damage repair mechanisms by BRCA1 is of great interest since molecular defects in this pathway may reflect a predictive value in terms of a cell’s sensitivity to DNA damaging agents or anticancer drugs. BRCA1 has been found to exhibit a hormone-dependent pattern of expression in breast cells. Wild-type BRCA1 is required for the inhibition of the growth of breast tumor cells in response to the pure steroidal ERα antagonist fulvestrant. Also a loss of BRCA1-mediated transcriptional activation of ERα expression results in increased resistance to ERα antagonists. Platinum-based drugs, poly(ADP-ribose) polymerase (PARP) inhibitors, and their combination are currently included in chemotherapy regimens for breast cancer. Preclinical and clinical studies in a BRCA1-defective setting have recently indicated a rationale for the use of these compounds against hereditary breast cancers. Initial findings indicate that neoadjuvant use of cisplatin results in high rates of complete pathological response in patients with breast cancer who have BRCA1 mutations. Cisplatin produces a better response in triple-negative breast cancer (TNBC) than in non-TNBC diseases in both the neoadjuvant and adjuvant settings. This implies that TNBC cells may harbor a dysfunctional BRCA1 repair pathway. Full article

(This article belongs to the Special Issue DNA Damage and Repair in Degenerative Diseases)

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12 pages, 926 KiB

Open AccessArticle

Low Temperature Hall Effect Investigation of Conducting Polymer-Carbon Nanotubes Composite Network

by Afarin Bahrami^1,2, Zainal Abidin Talib^1,*, Wan Mahmood Mat Yunus¹, Kasra Behzad^1,3, Mahnaz M.Abdi⁴ and Fasih Ud Din¹

¹ Department of Physics, Faculty of Science, University Putra Malaysia, Serdang 43400, Malaysia

² Department of Physics, Faculty of Science, Islamic Azad University, Eslamshahr Branch, 3314767653, Iran

³ Department of Physics, Faculty of Science, Islamic Azad University, Shahre Ghods Branch, 3754198811, Iran

⁴ Department of Chemistry, Faculty of Science, University Putra Malaysia, Serdang 43400, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 14917-14928; https://doi.org/10.3390/ijms131114917 - 14 Nov 2012

Cited by 23 | Viewed by 7132

Abstract

Polypyrrole (PPy) and polypyrrole-carboxylic functionalized multi wall carbon nanotube composites (PPy/f-MWCNT) were synthesized by in situ chemical oxidative polymerization of pyrrole on the carbon nanotubes (CNTs). The structure of the resulting complex nanotubes was characterized by transmission electron microscopy (TEM) and [...] Read more.

Polypyrrole (PPy) and polypyrrole-carboxylic functionalized multi wall carbon nanotube composites (PPy/f-MWCNT) were synthesized by in situ chemical oxidative polymerization of pyrrole on the carbon nanotubes (CNTs). The structure of the resulting complex nanotubes was characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). The effects of f-MWCNT concentration on the electrical properties of the resulting composites were studied at temperatures between 100 K and 300 K. The Hall mobility and Hall coefficient of PPy and PPy/f-MWCNT composite samples with different concentrations of f-MWCNT were measured using the van der Pauw technique. The mobility decreased slightly with increasing temperature, while the conductivity was dominated by the gradually increasing carrier density. Full article

(This article belongs to the Section Materials Science)

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17 pages, 756 KiB

Open AccessReview

Probing Nucleic Acid Interactions and Pre-mRNA Splicing by Förster Resonance Energy Transfer (FRET) Microscopy

by Eva Šimková and David Staněk^*

Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic

Int. J. Mol. Sci. 2012, 13(11), 14929-14945; https://doi.org/10.3390/ijms131114929 - 14 Nov 2012

Cited by 12 | Viewed by 12969

Abstract

Förster resonance energy transfer (FRET) microscopy is a powerful technique routinely used to monitor interactions between biomolecules. Here, we focus on the techniques that are used for investigating the structure and interactions of nucleic acids (NAs). We present a brief overview of the [...] Read more.

Förster resonance energy transfer (FRET) microscopy is a powerful technique routinely used to monitor interactions between biomolecules. Here, we focus on the techniques that are used for investigating the structure and interactions of nucleic acids (NAs). We present a brief overview of the most commonly used FRET microscopy techniques, their advantages and drawbacks. We list experimental approaches recently used for either in vitro or in vivo studies. Next, we summarize how FRET contributed to the understanding of pre-mRNA splicing and spliceosome assembly. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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10 pages, 201 KiB

Open AccessArticle

Rapid Development of Microsatellite Markers for the Endangered Fish Schizothorax biddulphi (Günther) Using Next Generation Sequencing and Cross-Species Amplification

by Wei Luo^1,†, Zhulan Nie^1,†, Fanbin Zhan¹, Jie Wei², Weimin Wang¹ and Zexia Gao^1,*

¹ Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China

² Key Laboratory of Tarim Animal Husbandry Science and Technology, College of Animal Science, Tarim University, Alar 843300, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 14946-14955; https://doi.org/10.3390/ijms131114946 - 14 Nov 2012

Cited by 41 | Viewed by 6967

Abstract

Tarim schizothoracin (Schizothorax biddulphi) is an endemic fish species native to the Tarim River system of Xinjiang and has been classified as an extremely endangered freshwater fish species in China. Here, we used a next generation sequencing platform (ion torrent PGM™) [...] Read more.

Tarim schizothoracin (Schizothorax biddulphi) is an endemic fish species native to the Tarim River system of Xinjiang and has been classified as an extremely endangered freshwater fish species in China. Here, we used a next generation sequencing platform (ion torrent PGM™) to obtain a large number of microsatellites for S. biddulphi, for the first time. A total of 40577 contigs were assembled, which contained 1379 SSRs. In these SSRs, the number of dinucleotide repeats were the most frequent (77.08%) and AC repeats were the most frequently occurring microsatellite, followed by AG, AAT and AT. Fifty loci were randomly selected for primer development; of these, 38 loci were successfully amplified and 29 loci were polymorphic across panels of 30 individuals. The H_o ranged from 0.15 to 0.83, and H_e ranged from 0.15 to 0.85, with 3.5 alleles per locus on average. Cross-species utility indicated that 20 of these markers were successfully amplified in a related, also an endangered fish species, S. irregularis. This study suggests that PGM™ sequencing is a rapid and cost-effective tool for developing microsatellite markers for non-model species and the developed microsatellite markers in this study would be useful in Schizothorax genetic analysis. Full article

(This article belongs to the Section Biochemistry)

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6 pages, 170 KiB

Open AccessTechnical Note

Polymorphisms in Myostatin Gene and Associations with Growth Traits in the Common Carp (Cyprinus carpio L.)

by Yanhong Sun^1,2, Xiaomu Yu¹ and Jingou Tong^1,*

¹ State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China

² Graduate School of Chinese Academy of Sciences, Beijing 100039, China

Int. J. Mol. Sci. 2012, 13(11), 14956-14961; https://doi.org/10.3390/ijms131114956 - 14 Nov 2012

Cited by 45 | Viewed by 5594

Abstract

Myostatin (MSTN) is a member of the transforming growth factor-β superfamily that negatively regulates skeletal muscle development and growth. In the present study, partial genomic fragments of MSTN were screened for single nucleotide polymorphisms (SNPs) in selected common carp individuals from [...] Read more.

Myostatin (MSTN) is a member of the transforming growth factor-β superfamily that negatively regulates skeletal muscle development and growth. In the present study, partial genomic fragments of MSTN were screened for single nucleotide polymorphisms (SNPs) in selected common carp individuals from wild populations, and two SNPs in intron 2 (c.371 + 749A > G, c.371 + 781T > C) and two synonymous SNPs in exon 3 (c.42A > G, c.72C > T) were identified. Genotyping by direct sequencing of polymerase chain reaction (PCR) products for these four SNPs were performed in 162 individuals from a commercial hatchery population. Association analysis showed that two SNPs in exon 3 were significantly associated with body weight (BW) and condition factor (K), and haplotype analyses revealed that haplotype H7H8 showed better growth performance. Our results demonstrated that some of the SNPs in MSTN may have positive effects on growth traits and suggested that MSTN could be a candidate gene for growth and marker-assisted selection in common carp. Full article

(This article belongs to the Section Biochemistry)

11 pages, 172 KiB

Open AccessArticle

Genomic Polymorphism of Human Papillomavirus Type 52 in Women from Northeast China

by Zhengrong Sun¹, Zhitao Lu¹, Jianhua Liu^1,2, Guili Wang¹, Weiqiang Zhou¹, Lianxia Yang¹, Chao Liu¹ and Qiang Ruan^1,*

¹ Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Heping District, Shenyang 110004, China

² Department of clinical laboratory, The Affiliated Shengjing Hospital, China Medical University, Heping District, Shenyang 110004, China

Int. J. Mol. Sci. 2012, 13(11), 14962-14972; https://doi.org/10.3390/ijms131114962 - 15 Nov 2012

Cited by 14 | Viewed by 6010

Abstract

Human papillomavirus (HPV) 52 is an oncogenic HPV type prevalent in Asia. The aim of the study was to analyze HPV 52 genetic variations in women from Northeast China. To explore the intratypic variants of HPV 52, the genomic regions of L1, [...] Read more.

Human papillomavirus (HPV) 52 is an oncogenic HPV type prevalent in Asia. The aim of the study was to analyze HPV 52 genetic variations in women from Northeast China. To explore the intratypic variants of HPV 52, the genomic regions of L1, E6, E7 and long control region (LCR) of HPV 52, which have been identified in women from Northeast China by HPV GenoArray test, were analyzed. Twenty-five mutations were identified in the regions examined. Of the mutations found in the L1 gene, three novel nonsynonymous mutations of C5640T, A5641T and G5642A were located within the region that encodes the binding domain of neutralizing antibodies against HPV 52. Although four variations were identified in HPV 52 E6 and E7 genes, no significant association was found between the mutations and the cytological lesion of the patients. Eight mutations, including a novel CTT7681–7683 deletion, found in the LCR of HPV 52 encompassed the known transcription binding sites, which may possibly affect the transcription of the oncogenic genes of E6 and E7. The most prevalent HPV 52 variant in women from northeastern China belongs to clade L1-LN-A. The genetic variations of HPV 52, including three novel nonsynonymous mutations of C5640T, A5641T and G5642A in the L1 gene and a novel CTT7681–7683 deletion in the LCR, were first documented in strains from women in Northeast China. The statistical result showed no associations between the variants and the severities of the infected women. These findings provide new data regarding gene variations of HPV 52. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

19 pages, 351 KiB

Open AccessArticle

Healing, Antioxidant and Cytoprotective Properties of Indigofera truxillensis in Different Models of Gastric Ulcer in Rats

by Anderson Luiz-Ferreira^1,2,*

, Maira Cola³, Victor Barbastefano², Felipe Meira De-Faria³, Ana Beatriz A. de Almeida², Elisângela Farias-Silva², Tamara Regina Calvo⁴, Clélia A. Hiruma-Lima⁵, Wagner Vilegas⁴

and Alba Regina M. Souza-Brito²

¹ Department of Biological Sciences, Federal University of Goiás, 75704-020, Catalão, Goiás, Brazil

² Department of Structural and Functional Biology, Biology Institute, Campinas University, 13083-865, Campinas, São Paulo, Brazil

³ Department of Pharmacology, Faculty of Medical Sciences, University of Campinas, 13083-887, Campinas, São Paulo, Brazil

⁴ Department of Organic Chemistry, Institute of Chemistry, São Paulo State University, 14800-900, Araraquara, São Paulo, Brazil

⁵ Department of Physiology, Institute of Biosciences, São Paulo State University, 18618-000, Botucatu, São Paulo, Brazil

Int. J. Mol. Sci. 2012, 13(11), 14973-14991; https://doi.org/10.3390/ijms131114973 - 15 Nov 2012

Cited by 17 | Viewed by 7301

Abstract

The present study evaluated the antiulcerogenic activity and mechanisms of the aqueous (AqF 100 mg/kg) and ethyl acetate (AcF 50 mg/kg) fractions from Indigofera truxillensis leaves. This dose was selected to assess its activity on ulcer healing and its action on gastric acid [...] Read more.

The present study evaluated the antiulcerogenic activity and mechanisms of the aqueous (AqF 100 mg/kg) and ethyl acetate (AcF 50 mg/kg) fractions from Indigofera truxillensis leaves. This dose was selected to assess its activity on ulcer healing and its action on gastric acid and mucus secretion, prostaglandin production and antioxidant enzyme activity (superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rd)). Gastric ulcer was induced by absolute ethanol. Antisecretory action, mucus and prostaglandin production, healing and antioxidant enzyme activities were evaluated for both fractions. AqF and AcF significantly inhibited the gastric mucosal damage caused by ethanol. This effect was statistically significant at 100 and 50 mg/kg compared with the vehicle. Neither fraction interfered with gastric secretion. AcF increased the PGE₂ production, and both fractions increased mucus production. l-NAME did not alter the gastroprotection exerted by the fractions, but N-ethylmaleimide attenuated only AcF. In the ischemia/reperfusion model both fractions inhibited the mucosal damage. AcF increased SOD, GSH-Px and GSH-Rd activity, but AqF increased only SOD and GSH-Px. In the acetic acid-induced ulcer model AcF only accelerated ulcer healing. These results showed that Indigofera truxillensis acted as a gastroprotective agent, stimulating protective factors and antioxidants enzymes. Full article

(This article belongs to the Section Biochemistry)

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20 pages, 2503 KiB

Open AccessArticle

Betulin Complex in γ-Cyclodextrin Derivatives: Properties and Antineoplasic Activities in In Vitro and In Vivo Tumor Models

by Codruta Şoica¹, Cristina Dehelean^1,*, Corina Danciu¹, Hai Ming Wang², Gerhard Wenz², Rita Ambrus³, Florina Bojin⁴

and Mariana Anghel⁴

¹ Faculty of Pharmacy, University of Medicine and Pharmacy, 2 Eftimie Murgu, Timisoara 300041, Romania

² Organic Macromolecular Chemistry, Saarland University, Campus Saarbrücken, Saarbrücken D-66123, Germany

³ Institute of Pharmaceutical Technology, University of Szeged, 6 Eotvos ut., Szeged H-6720, Hungary

⁴ Faculty of Medicine, University of Medicine and Pharmacy, 2 Eftimie Murgu, Timisoara 300041, Romania

Int. J. Mol. Sci. 2012, 13(11), 14992-15011; https://doi.org/10.3390/ijms131114992 - 15 Nov 2012

Cited by 70 | Viewed by 9609

Abstract

Given the present high incidence of melanoma and skin cancer, interest in potential drugs of plant origin has increased significantly. Pentacyclic lupane-type triterpenes are widely distributed in plants, offering numerous pharmacological benefits. Betulin is an important compound in the bark of Betula pendula [...] Read more.

Given the present high incidence of melanoma and skin cancer, interest in potential drugs of plant origin has increased significantly. Pentacyclic lupane-type triterpenes are widely distributed in plants, offering numerous pharmacological benefits. Betulin is an important compound in the bark of Betula pendula Roth and has important therapeutic properties, including antitumor activities. Its biological effect is limited by its poor water solubility, which can be improved by cyclodextrin complexation. The best results have been obtained by using a novel cyclodextrin derivative, octakis-[6-deoxy-6-(2-sulfanyl ethanesulfonate)]-γ-CD. The complexes between betulin and the previously mentioned cyclodextrin were analyzed by scanning electron microscopy (SEM)and differential scanning calorimetry (DSC) and pharmacologically evaluated in vitro (MTT and immunocytochemistry tests) and in vivo in C57BL/6J mice. The solubility of betulin is improved by cyclodextrin complexation, which creates a stable complex that improves the in vitro and in vivo properties of the active compound. Full article

(This article belongs to the Section Materials Science)

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30 pages, 1408 KiB

Open AccessReview

Dynamic Control of Electron Transfers in Diflavin Reductases

by Louise Aigrain¹, Fataneh Fatemi², Oriane Frances², Ewen Lescop² and Gilles Truan^3,4,5,*

¹ Gene Machines Group, Clarendon Laboratory, Department of Physics, University of Oxford, Parks Road, Oxford OX1 3PU, UK

² Institut de Chimie des Substances Naturelles, CNRS, UPR 2301, Centre de Recherche de Gif, 1 Av. de la Terrasse, 91198 Gif-sur-Yvette Cedex, France

³ Université de Toulouse; INSA, UPS, INP; LISBP, 135 Avenue de Rangueil, F-31077 Toulouse, France

⁴ Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse, France

⁵ CNRS, UMR5504, F-31400 Toulouse, France

Int. J. Mol. Sci. 2012, 13(11), 15012-15041; https://doi.org/10.3390/ijms131115012 - 15 Nov 2012

Cited by 27 | Viewed by 8259

Abstract

Diflavin reductases are essential proteins capable of splitting the two-electron flux from reduced pyridine nucleotides to a variety of one electron acceptors. The primary sequence of diflavin reductases shows a conserved domain organization harboring two catalytic domains bound to the FAD and FMN [...] Read more.

Diflavin reductases are essential proteins capable of splitting the two-electron flux from reduced pyridine nucleotides to a variety of one electron acceptors. The primary sequence of diflavin reductases shows a conserved domain organization harboring two catalytic domains bound to the FAD and FMN flavins sandwiched by one or several non-catalytic domains. The catalytic domains are analogous to existing globular proteins: the FMN domain is analogous to flavodoxins while the FAD domain resembles ferredoxin reductases. The first structural determination of one member of the diflavin reductases family raised some questions about the architecture of the enzyme during catalysis: both FMN and FAD were in perfect position for interflavin transfers but the steric hindrance of the FAD domain rapidly prompted more complex hypotheses on the possible mechanisms for the electron transfer from FMN to external acceptors. Hypotheses of domain reorganization during catalysis in the context of the different members of this family were given by many groups during the past twenty years. This review will address the recent advances in various structural approaches that have highlighted specific dynamic features of diflavin reductases. Full article

(This article belongs to the Special Issue Flavins)

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12 pages, 2758 KiB

Open AccessArticle

Antifungal Activity of (KW)_n or (RW)_n Peptide against Fusarium solani and Fusarium oxysporum

by Ramamourthy Gopal¹, Hyungjong Na², Chang Ho Seo³ and Yoonkyung Park^1,2,*

¹ Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Korea

² Department of Biotechnology, Chosun University, Gwangju 501-759, Korea

³ Department of Bioinformatics, Kongju National University, Kongju 314-701, Korea

Int. J. Mol. Sci. 2012, 13(11), 15042-15053; https://doi.org/10.3390/ijms131115042 - 15 Nov 2012

Cited by 31 | Viewed by 8138

Abstract

The presence of lysine (Lys) or arginine (Arg) and tryptophan (Trp) are important for the antimicrobial effects of cationic peptides. Therefore, we designed and synthesized a series of antimicrobial peptides with various numbers of Lys (or Arg) and Trp repeats [(KW and RW) [...] Read more.

The presence of lysine (Lys) or arginine (Arg) and tryptophan (Trp) are important for the antimicrobial effects of cationic peptides. Therefore, we designed and synthesized a series of antimicrobial peptides with various numbers of Lys (or Arg) and Trp repeats [(KW and RW)_n-NH₂, where n equals 2, 3, 4, or 5]. Antifungal activities of these peptides increased with chain length. Light microscopy demonstrated that longer peptides (n = 4, 5) strongly inhibited in vitro growth of Fusarium solani, and Fusarium oxysporum, at 4–32 μM. Furthermore, longer peptides displayed potent fungicidal activities against a variety of agronomical important filamentous fungi, including F. solani and F. oxysporum, at their minimal inhibitory concentrations (MICs). However, RW series peptides showed slightly higher fungicidal activities than KW peptides against the two strains. Taken together, the results of this study indicate that these short peptides would be good candidates for use as synthetic or transgenic antifungal agents. Full article

(This article belongs to the Special Issue Green Biocides)

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20 pages, 257 KiB

Open AccessReview

A Review of Molecular Mechanisms of the Anti-Leukemic Effects of Phenolic Compounds in Honey

by Murtala B. Abubakar^1,*, Wan Zaidah Abdullah², Siti Amrah Sulaiman³ and Ang Boon Suen¹

¹ Department of Physiology, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

² Department of Haematology, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

³ Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 15054-15073; https://doi.org/10.3390/ijms131115054 - 15 Nov 2012

Cited by 78 | Viewed by 12375

Abstract

Hematologic malignancies constitute about 9% of all new cases of cancers as reported via the GLOBOCAN series by International Agency for Research on Cancer (IARC) in 2008. So far, the conventional therapeutic and surgical approaches to cancer therapy have not been able to [...] Read more.

Hematologic malignancies constitute about 9% of all new cases of cancers as reported via the GLOBOCAN series by International Agency for Research on Cancer (IARC) in 2008. So far, the conventional therapeutic and surgical approaches to cancer therapy have not been able to curtail the rising incidence of cancers, including hematological malignancies, worldwide. The last decade has witnessed great research interest in biological activities of phenolic compounds that include anticancer, anti-oxidation and anti-inflammation, among other things. A large number of anticancer agents combat cancer through cell cycle arrest, induction of apoptosis and differentiation, as well as through inhibition of cell growth and proliferation, or a combination of two or more of these mechanisms. Various phenolic compounds from different sources have been reported to be promising anticancer agents by acting through one of these mechanisms. Honey, which has a long history of human consumption both for medicinal and nutritional uses, contains a variety of phenolic compounds such as flavonoids, phenolic acids, coumarins and tannins. This paper presents a review on the molecular mechanisms of the anti-leukemic activity of various phenolic compounds on cell cycle, cell growth and proliferation and apoptosis, and it advocates that more studies should be conducted to determine the potential role of honey in both chemoprevention and chemotherapy in leukemia. Full article

(This article belongs to the Section Biochemistry)

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12 pages, 605 KiB

Open AccessArticle

Histone Deacetylase (HDAC) Inhibitors Down-Regulate Endothelial Lineage Commitment of Umbilical Cord Blood Derived Endothelial Progenitor Cells

by Florin Iordache, Cosmin Buzila, Andrei Constantinescu, Eugen Andrei and Horia Maniu^*

Institute of Cellular Biology and Pathology of Romanian Academy, “Nicolae Simionescu”, 8, B.P. Hasdeu, Bucharest 050568, Romania

Int. J. Mol. Sci. 2012, 13(11), 15074-15085; https://doi.org/10.3390/ijms131115074 - 15 Nov 2012

Cited by 19 | Viewed by 7374

Abstract

To test the involvement of histone deacetylases (HDACs) activity in endothelial lineage progression, we investigated the effects of HDAC inhibitors on endothelial progenitors cells (EPCs) derived from umbilical cord blood (UCB). Adherent EPCs, that expressed the endothelial marker proteins (PCAM-1, CD105, CD133, and [...] Read more.

To test the involvement of histone deacetylases (HDACs) activity in endothelial lineage progression, we investigated the effects of HDAC inhibitors on endothelial progenitors cells (EPCs) derived from umbilical cord blood (UCB). Adherent EPCs, that expressed the endothelial marker proteins (PCAM-1, CD105, CD133, and VEGFR₂) revealed by flow cytometry were treated with three HDAC inhibitors: Butyrate (BuA), Trichostatin A (TSA), and Valproic acid (VPA). RT-PCR assay showed that HDAC inhibitors down-regulated the expression of endothelial genes such as VE-cadherin, CD133, CXCR4 and Tie-2. Furthermore, flow cytometry analysis illustrated that HDAC inhibitors selectively reduce the expression of VEGFR₂, CD117, VE-cadherin, and ICAM-1, whereas the expression of CD34 and CD45 remained unchanged, demonstrating that HDAC is involved in endothelial differentiation of progenitor cells. Real-Time PCR demonstrated that TSA down-regulated telomerase activity probably via suppression of hTERT expression, suggesting that HDAC inhibitor decreased cell proliferation. Cell motility was also decreased after treatment with HDAC inhibitors as shown by wound-healing assay. The balance of acethylation/deacethylation kept in control by the activity of HAT (histone acetyltransferases)/HDAC enzymes play an important role in differentiation of stem cells by regulating proliferation and endothelial lineage commitment. Full article

(This article belongs to the Section Biochemistry)

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21 pages, 353 KiB

Open AccessReview

Bacterial Bio-Resources for Remediation of Hexachlorocyclohexane

by Analía Alvarez^1,2,†, Claudia S. Benimeli^1,3,†, Juliana M. Saez¹, María S. Fuentes¹, Sergio A. Cuozzo^1,2, Marta A. Polti^1,2 and María J. Amoroso^1,3,4,*

¹ Pilot Plant of Industrial and Microbiological Processes (PROIMI), CONICET, Av. Belgrano y Pasaje Caseros, 4000 Tucumán, Argentina

² Natural Sciences College and Miguel Lillo Institute, National University of Tucumán, Miguel Lillo 205, 4000 Tucumán, Argentina

³ North University of Saint Thomas Aquines, 9 de Julio 165, 4000 Tucumán, Argentina

⁴ Biochemistry, Chemistry and Pharmacy College, National University of Tucumán, Ayacucho 471, 4000 Tucumán, Argentina

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 15086-15106; https://doi.org/10.3390/ijms131115086 - 15 Nov 2012

Cited by 68 | Viewed by 10265

Abstract

In the last few decades, highly toxic organic compounds like the organochlorine pesticide (OP) hexachlorocyclohexane (HCH) have been released into the environment. All HCH isomers are acutely toxic to mammals. Although nowadays its use is restricted or completely banned in most countries, it [...] Read more.

In the last few decades, highly toxic organic compounds like the organochlorine pesticide (OP) hexachlorocyclohexane (HCH) have been released into the environment. All HCH isomers are acutely toxic to mammals. Although nowadays its use is restricted or completely banned in most countries, it continues posing serious environmental and health concerns. Since HCH toxicity is well known, it is imperative to develop methods to remove it from the environment. Bioremediation technologies, which use microorganisms and/or plants to degrade toxic contaminants, have become the focus of interest. Microorganisms play a significant role in the transformation and degradation of xenobiotic compounds. Many Gram-negative bacteria have been reported to have metabolic abilities to attack HCH. For instance, several Sphingomonas strains have been reported to degrade the pesticide. On the other hand, among Gram-positive microorganisms, actinobacteria have a great potential for biodegradation of organic and inorganic toxic compounds. This review compiles and updates the information available on bacterial removal of HCH, particularly by Streptomyces strains, a prolific genus of actinobacteria. A brief account on the persistence and deleterious effects of these pollutant chemical is also given. Full article

(This article belongs to the Special Issue Green Biocides)

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19 pages, 201 KiB

Open AccessReview

Mast Cells in the Pathogenesis of Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis

by Massimo Costanza¹, Mario P. Colombo² and Rosetta Pedotti^1,*

¹ Neuroimmunology and Neuromuscular Disorder Unit, Neurological Institute Foundation IRCCS C. Besta, via Amadeo 42, Milan 20133, Italy

² Molecular Immunology Unit, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, via Amadeo 42, Milan 20133, Italy

Int. J. Mol. Sci. 2012, 13(11), 15107-15125; https://doi.org/10.3390/ijms131115107 - 16 Nov 2012

Cited by 32 | Viewed by 8642

Abstract

Mast cells (MCs) are best known as key immune players in immunoglobulin E (IgE)-dependent allergic reactions. In recent years, several lines of evidence have suggested that MCs might play an important role in several pathological conditions, including autoimmune disorders such as multiple sclerosis [...] Read more.

Mast cells (MCs) are best known as key immune players in immunoglobulin E (IgE)-dependent allergic reactions. In recent years, several lines of evidence have suggested that MCs might play an important role in several pathological conditions, including autoimmune disorders such as multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), an animal model for MS. Since their first description in MS plaques in the late 1800s, much effort has been put into elucidating the contribution of MCs to the development of central nervous system (CNS) autoimmunity. Mouse models of MC-deficiency have provided a valuable experimental tool for dissecting MC involvement in MS and EAE. However, to date there is still major controversy concerning the function of MCs in these diseases. Indeed, although MCs have been classically proposed as having a detrimental and pro-inflammatory role, recent literature has questioned and resized the contribution of MCs to the pathology of MS and EAE. In this review, we will present the main evidence obtained in MS and EAE on this topic, and discuss the critical and controversial aspects of such evidence. Full article

(This article belongs to the Special Issue Recent Advances in the Research of Multiple Sclerosis)

11 pages, 284 KiB

Open AccessArticle

Catalytic Hydrogenation of the Sweet Principles of Stevia rebaudiana, Rebaudioside B, Rebaudioside C, and Rebaudioside D and Sensory Evaluation of Their Reduced Derivatives

by Indra Prakash¹, Mary Campbell² and Venkata Sai Prakash Chaturvedula^1,*

¹ Organic Chemistry Department, Global Research and Development, The Coca-Cola Company, One Coca-Cola Plaza, Atlanta, GA 30313, USA

² Product Development, Coca-Cola North America Division, The Coca-Cola Company, One Coca-Cola Plaza, Atlanta, GA 30313, USA

Int. J. Mol. Sci. 2012, 13(11), 15126-15136; https://doi.org/10.3390/ijms131115126 - 16 Nov 2012

Cited by 30 | Viewed by 8632

Abstract

Catalytic hydrogenation of rebaudioside B, rebaudioside C, and rebaudioside D; the three ent-kaurane diterpene glycosides isolated from Stevia rebaudiana was carried out using Pd(OH)₂. Reduction of steviol glycosides was performed using straightforward synthetic chemistry with the catalyst Pd(OH)₂ and [...] Read more.

Catalytic hydrogenation of rebaudioside B, rebaudioside C, and rebaudioside D; the three ent-kaurane diterpene glycosides isolated from Stevia rebaudiana was carried out using Pd(OH)₂. Reduction of steviol glycosides was performed using straightforward synthetic chemistry with the catalyst Pd(OH)₂ and structures of the corresponding dihydro derivatives were characterized on the basis of 1D and 2D nuclear magnetic resonance (NMR) spectral data indicating that all are novel compounds being reported for the first time. Also, the taste properties of all reduced compounds were evaluated against their corresponding original steviol glycosides and sucrose. Full article

(This article belongs to the Section Biochemistry)

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25 pages, 869 KiB

Open AccessReview

Förster Resonance Energy Transfer (FRET) as a Tool for Dissecting the Molecular Mechanisms for Maturation of the Shigella Type III Secretion Needle Tip Complex

by Nicholas E. Dickenson^* and William D. Picking

Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK 74078, USA

Int. J. Mol. Sci. 2012, 13(11), 15137-15161; https://doi.org/10.3390/ijms131115137 - 16 Nov 2012

Cited by 13 | Viewed by 7350

Abstract

Förster resonance energy transfer (FRET) provides a powerful tool for monitoring intermolecular interactions and a sensitive technique for studying Å-level protein conformational changes. One system that has particularly benefited from the sensitivity and diversity of FRET measurements is the maturation of the Shigella [...] Read more.

Förster resonance energy transfer (FRET) provides a powerful tool for monitoring intermolecular interactions and a sensitive technique for studying Å-level protein conformational changes. One system that has particularly benefited from the sensitivity and diversity of FRET measurements is the maturation of the Shigella type III secretion apparatus (T3SA) needle tip complex. The Shigella T3SA delivers effector proteins into intestinal cells to promote bacterial invasion and spread. The T3SA is comprised of a basal body that spans the bacterial envelope and a needle with an exposed tip complex that matures in response to environmental stimuli. FRET measurements demonstrated bile salt binding by the nascent needle tip protein IpaD and also mapped resulting structural changes which led to the recruitment of the translocator IpaB. At the needle tip IpaB acts as a sensor for host cell contact but prior to secretion, it is stored as a heterodimeric complex with the chaperone IpgC. FRET analyses showed that chaperone binding to IpaB’s N-terminal domain causes a conformational change in the latter. These FRET analyses, with other biophysical methods, have been central to understanding T3SA maturation and will be highlighted, focusing on the details of the FRET measurements and the relevance to this particular system. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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15 pages, 639 KiB

Open AccessArticle

The Characterization of SaPIN2b, a Plant Trichome-Localized Proteinase Inhibitor from Solanum americanum

by Ming Luo^1,2,†, Ling-Wen Ding^1,†, Zhi-Juan Ge^1,†, Zhen-Yu Wang¹, Bo-Lun Hu¹, Xiao-Bei Yang¹, Qiao-Yang Sun¹ and Zeng-Fu Xu^1,3,*

¹ State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China

² Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China

³ Key Laboratory of Tropical Plant Resource and Sustainable Use, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Menglun, Yunnan 666303, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 15162-15176; https://doi.org/10.3390/ijms131115162 - 16 Nov 2012

Cited by 17 | Viewed by 7147

Abstract

Proteinase inhibitors play an important role in plant resistance of insects and pathogens. In this study, we characterized the serine proteinase inhibitor SaPIN2b, which is constitutively expressed in Solanum americanum trichomes and contains two conserved motifs of the proteinase inhibitor II (PIN2) family. [...] Read more.

Proteinase inhibitors play an important role in plant resistance of insects and pathogens. In this study, we characterized the serine proteinase inhibitor SaPIN2b, which is constitutively expressed in Solanum americanum trichomes and contains two conserved motifs of the proteinase inhibitor II (PIN2) family. The recombinant SaPIN2b (rSaPIN2b), which was expressed in Escherichia coli, was demonstrated to be a potent proteinase inhibitor against a panel of serine proteinases, including subtilisin A, chymotrypsin and trypsin. Moreover, rSaPIN2b also effectively inhibited the proteinase activities of midgut trypsin-like proteinases that were extracted from the devastating pest Helicoverpa armigera. Furthermore, the overexpression of SaPIN2b in transgenic tobacco plants resulted in enhanced resistance against H. armigera. Taken together, our results demonstrated that SaPIN2b is a potent serine proteinase inhibitor that may act as a protective protein in plant defense against insect attacks. Full article

(This article belongs to the Special Issue Advances in Molecular Plant Biology)

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16 pages, 515 KiB

Open AccessArticle

Detection of Glycomic Alterations Induced by Overexpression of P-Glycoprotein on the Surfaces of L1210 Cells Using Sialic Acid Binding Lectins

by Tatiana Bubencíkova¹, Dana Cholujová², Lucia Messingerová¹, Danica Mislovicova³, Mario Seres¹, Albert Breier^1,* and Zdena Sulova^1,*

¹ Institute of Molecular Physiology and Genetics, Center of Excellence of the Slovak Research and Development Agency “BIOMEMBRANES2008”, Slovak Academy of Sciences, Vlarska 5, Bratislava 83334, Slovakia

² Cancer Research Institute, Slovak Academy of Sciences, Vlarska 7, Bratislava 83391, Slovakia

³ Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava 84538, Slovakia

Int. J. Mol. Sci. 2012, 13(11), 15177-15192; https://doi.org/10.3390/ijms131115177 - 16 Nov 2012

Cited by 13 | Viewed by 7262

Abstract

P-glycoprotein (P-gp) overexpression is the most frequently observed cause of multidrug resistance in neoplastic cells. In our experiments, P-gp was expressed in L1210 mice leukemia cells (S cells) by selection with vincristine (R cells) or transfection with the gene encoding human P-gp (T [...] Read more.

P-glycoprotein (P-gp) overexpression is the most frequently observed cause of multidrug resistance in neoplastic cells. In our experiments, P-gp was expressed in L1210 mice leukemia cells (S cells) by selection with vincristine (R cells) or transfection with the gene encoding human P-gp (T cells). Remodeling of cell surface sugars is associated with P-gp expression in L1210 cells as a secondary cellular response. In this study, we monitored the alteration of cell surface saccharides by Sambucus nigra agglutinin (SNA), wheat germ agglutinin (WGA) and Maackia amurensis agglutinin (MAA). Sialic acid is predominantly linked to the surface of S, R and T cells via α-2,6 branched sugars that tightly bind SNA. The presence of sialic acid linked to the cell surface via α-2,3 branched sugars was negligible, and the binding of MAA (recognizing this branch) was much less pronounced than SNA. WGA induced greater cell death than SNA, which was bound to the cell surface and agglutinated all three L1210 cell-variants more effectively than WGA. Thus, the ability of lectins to induce cell death did not correlate with their binding efficiency and agglutination potency. Compared to S cells, P-gp positive R and T cells contain a higher amount of N-acetyl-glucosamine on their cell surface, which is associated with improved WGA binding. Both P-gp positive variants of L1210 cells are strongly resistant to vincristine as P-gp prototypical drug. This resistance could not be altered by liberalization of terminal sialyl residues from the cell surface by sialidase. Full article

(This article belongs to the Section Molecular Toxicology)

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16 pages, 249 KiB

Open AccessArticle

Nitric Oxide-Dependent Posttranslational Modification in Plants: An Update

by Jeremy Astier^* and Christian Lindermayr^*

Institute of Biochemical Plant Pathology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany

Int. J. Mol. Sci. 2012, 13(11), 15193-15208; https://doi.org/10.3390/ijms131115193 - 16 Nov 2012

Cited by 190 | Viewed by 11875

Abstract

Nitric oxide (NO) has been demonstrated as an essential regulator of several physiological processes in plants. The understanding of the molecular mechanism underlying its critical role constitutes a major field of research. NO can exert its biological function through different ways, such as [...] Read more.

Nitric oxide (NO) has been demonstrated as an essential regulator of several physiological processes in plants. The understanding of the molecular mechanism underlying its critical role constitutes a major field of research. NO can exert its biological function through different ways, such as the modulation of gene expression, the mobilization of second messengers, or interplays with protein kinases. Besides this signaling events, NO can be responsible of the posttranslational modifications (PTM) of target proteins. Several modifications have been identified so far, whereas metal nitrosylation, the tyrosine nitration and the S-nitrosylation can be considered as the main ones. Recent data demonstrate that these PTM are involved in the control of a wide range of physiological processes in plants, such as the plant immune system. However, a great deal of effort is still necessary to pinpoint the role of each PTM in plant physiology. Taken together, these new advances in proteomic research provide a better comprehension of the role of NO in plant signaling. Full article

(This article belongs to the Special Issue Advances in Proteomic Research)

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18 pages, 589 KiB

Open AccessArticle

Isolation and Structural Characterization of Lignin from Cotton Stalk Treated in an Ammonia Hydrothermal System

by Sumin Kang¹, Lingping Xiao¹

, Lingyan Meng¹, Xueming Zhang^1,*

and Runcang Sun^1,2,*

¹ Institute of Biomass Chemistry and Technology, Beijing Forestry University, Beijing 100083, China

² State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510640, China

Int. J. Mol. Sci. 2012, 13(11), 15209-15226; https://doi.org/10.3390/ijms131115209 - 16 Nov 2012

Cited by 73 | Viewed by 8371

Abstract

To investigate the potential for the utilization of cotton stalk, ammonia hydrothermal treatment was applied to fractionate the samples into aqueous ammonia-soluble and ammonia-insoluble portions. The ammonia-soluble portion was purified to yield lignin fractions. The lignin fractions obtained were characterized by wet chemistry [...] Read more.

To investigate the potential for the utilization of cotton stalk, ammonia hydrothermal treatment was applied to fractionate the samples into aqueous ammonia-soluble and ammonia-insoluble portions. The ammonia-soluble portion was purified to yield lignin fractions. The lignin fractions obtained were characterized by wet chemistry (carbohydrate analysis) and spectroscopy methods (FT-IR, ¹³C and ¹H-¹³C HSQC NMR spectroscopy) as well as gel permeation chromatography (GPC). The results showed that the cotton stalk lignin fractions were almost absent of neutral sugars (0.43%–1.29%) and had relatively low average molecular weights (1255–1746 g/mol). The lignin fractions belonged to typical G-S lignin, which was composed predominately of G-type units (59%) and noticeable amounts of S-type units (40%) together with a small amount of H-type units (~1%). Furthermore, the ammonia-extractable lignin fractions were mainly composed of β-O-4' inter-unit linkages (75.6%), and small quantities of β-β' (12.2%), together with lower amounts of β-5' carbon-carbon linkages (7.4%) and p-hydroxycinnamyl alcohol end groups. Full article

(This article belongs to the Section Materials Science)

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14 pages, 1162 KiB

Open AccessArticle

Modification of Förster Resonance Energy Transfer Efficiencyat Interfaces

by Jörg Enderlein

III. Institute of Physics – Biophysics, Georg August University, Friedrich-Hund-Platz 1, D-37077 Göttingen, Germany

Int. J. Mol. Sci. 2012, 13(11), 15227-15240; https://doi.org/10.3390/ijms131115227 - 19 Nov 2012

Cited by 14 | Viewed by 6693

Abstract

We present a theoretical study on the impact of an interface on the FRET efficiency of a surface-bound acceptor-donor system. The FRET efficiency can be modified by two effects. Firstly, the donor’s electromagnetic field at the acceptor’s position is changed due to the [...] Read more.

We present a theoretical study on the impact of an interface on the FRET efficiency of a surface-bound acceptor-donor system. The FRET efficiency can be modified by two effects. Firstly, the donor’s electromagnetic field at the acceptor’s position is changed due to the partial reflection of the donor’s field. Secondly, both the donor’s and the acceptor’s quantum yield of fluorescence can be changed due to the interface-induced enhancement of the radiative emission rate (Purcell effect). Numerical results for a FRET-pair at a glass-water interface are given. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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11 pages, 227 KiB

Open AccessArticle

Synthesis Method for Thiosulfonate and Report of Its Insecticidal Activity in Anagasta kuehniella (Lepidoptera: Pyralidae)

by Edson Dos A. Dos Santos¹, Fernando M. Gonçalves², Paulo César Prado¹, Daniele Y. Sasaki², Dênis P. De Lima¹ and Maria Lígia Rodrigues Macedo^2,*

¹ Laboratory of Synthesis and Transformation of Organic Molecules, Chemistry Section of CCET, Federal University of Mato Grosso do Sul, CP 549, CEP 79070-900, Campo Grande-MS, Brazil

² Laboratory of Protein Purification and its Biological Functions, CCBS, Federal University of Mato Grosso do Sul, CP 549, CEP 79070-900, Campo Grande-MS, Brazil

Int. J. Mol. Sci. 2012, 13(11), 15241-15251; https://doi.org/10.3390/ijms131115241 - 19 Nov 2012

Cited by 19 | Viewed by 6734

Abstract

Insect pests have caused economic losses valued at billions of dollars in agricultural production. Anagasta kuehniella (Zeller), the Mediterranean flour moth, is of major economic importance as a flour and grain feeder and is often a severe pest in flourmills. This study provides [...] Read more.

Insect pests have caused economic losses valued at billions of dollars in agricultural production. Anagasta kuehniella (Zeller), the Mediterranean flour moth, is of major economic importance as a flour and grain feeder and is often a severe pest in flourmills. This study provides a suitable route for the direct preparation of thiosulfonates 2 and 3 from thiols, under mild conditions, with good yields; these thiosulfonates were tested for their regulatory effect on insect growth. The chronic ingestion of thiosulfonates resulted in a significant reduction in larval survival and weight. In addition, the tryptic activity of larvae was sensitive to these thiosulfonates. Results suggest that thiosulfonates 2 and 3 have a potential antimetabolic effect when ingested by A. kuehniella. The use of AgNO₃/BF₃·OEt₂ and Al(H₂PO₄)₃/HNO₃ provides a suitable route for the direct preparation of thiosulfonates from thiols under mild conditions with good yields. These thiosulfonates were toxic for A. kuehniella larvae, suggesting their potential as biotechnological tools. Full article

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19 pages, 368 KiB

Open AccessArticle

Simple Estimation of Förster Resonance Energy Transfer (FRET) Orientation Factor Distribution in Membranes

by Luís M. S. Loura^1,2

¹ Faculty of Pharmacy, University of Coimbra, Health Sciences Campus, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal

² Centre for Chemistry-Coimbra, Rua Larga, 3004-535 Coimbra, Portugal

Int. J. Mol. Sci. 2012, 13(11), 15252-15270; https://doi.org/10.3390/ijms131115252 - 19 Nov 2012

Cited by 65 | Viewed by 9760

Abstract

Because of its acute sensitivity to distance in the nanometer scale, Förster resonance energy transfer (FRET) has found a large variety of applications in many fields of chemistry, physics, and biology. One important issue regarding the correct usage of FRET is its dependence [...] Read more.

Because of its acute sensitivity to distance in the nanometer scale, Förster resonance energy transfer (FRET) has found a large variety of applications in many fields of chemistry, physics, and biology. One important issue regarding the correct usage of FRET is its dependence on the donor-acceptor relative orientation, expressed as the orientation factor κ². Different donor/acceptor conformations can lead to κ² values in the 0 ≤ κ² ≤ 4 range. Because the characteristic distance for FRET, R₀, is proportional to (κ²)^1/6, uncertainties in the orientation factor are reflected in the quality of information that can be retrieved from a FRET experiment. In most cases, the average value of κ² corresponding to the dynamic isotropic limit (<κ²> = 2/3) is used for computation of R₀ and hence donor-acceptor distances and acceptor concentrations. However, this can lead to significant error in unfavorable cases. This issue is more critical in membrane systems, because of their intrinsically anisotropic nature and their reduced fluidity in comparison to most common solvents. Here, a simple numerical simulation method for estimation of the probability density function of κ² for membrane-embedded donor and acceptor fluorophores in the dynamic regime is presented. In the simplest form, the proposed procedure uses as input the most probable orientations of the donor and acceptor transition dipoles, obtained by experimental (including linear dichroism) or theoretical (such as molecular dynamics simulation) techniques. Optionally, information about the widths of the donor and/or acceptor angular distributions may be incorporated. The methodology is illustrated for special limiting cases and common membrane FRET pairs. Full article

(This article belongs to the Special Issue Förster Resonance Energy Transfer (FRET))

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8 pages, 225 KiB

Open AccessReview

Role of Oxidative Stress in Hepatocarcinogenesis Induced by Hepatitis C Virus

by Kyoko Tsukiyama-Kohara

Department of Animal Hygiene, Transboundary Animal Diseases Center, Joint Faculty of Veterinary Medicine Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan

Int. J. Mol. Sci. 2012, 13(11), 15271-15278; https://doi.org/10.3390/ijms131115271 - 19 Nov 2012

Cited by 37 | Viewed by 8402

Abstract

Hepatitis C virus (HCV) easily establishes chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). During the progression of HCV infections, reactive oxygen species (ROS) are generated, and these ROS then induce significant DNA damage. The role of ROS in the pathogenesis of HCV infection [...] Read more.

Hepatitis C virus (HCV) easily establishes chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). During the progression of HCV infections, reactive oxygen species (ROS) are generated, and these ROS then induce significant DNA damage. The role of ROS in the pathogenesis of HCV infection is still not fully understood. Recently, we found that HCV induced the expression of 3β-hydroxysterol ∆24-reductase (DHCR24). We also found that a HCV responsive region is present in the 5'-flanking genomic promoter region of DHCR24 and the HCV responsive region was characterized as (−167/−140). Moreover, the transcription factor Sp1 was found to bind to this region in response to oxidative stress under the regulation of ataxia telangiectasia mutated (ATM) kinase. Overexpression of DHCR24 impaired p53 activity by suppression of acetylation and increased interaction with MDM2. This impairment of p53 suppressed the hydrogen peroxide-induced apoptotic response in hepatocytes. Thus, a target of oxidative stress in HCV infection is DHCR24 through Sp1, which suppresses apoptotic responses and increases tumorigenicity. Full article

(This article belongs to the Special Issue Oxidative Stress and Ageing)

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12 pages, 2789 KiB

Open AccessArticle

Molecular Mechanisms of RADA16-1 Peptide on Fast Stop Bleeding in Rat Models

by Ting Wang¹, Xiaozhong Zhong², Songtao Wang^1,3, Fei Lv¹ and Xiaojun Zhao^1,*

¹ Institute for NanoBiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan, China

² College of Life Science, Jianghan University, Wuhan 430056, Hubei, China

³ National Engineering Research Center for solid state fermentation, Luzhou 640040, Sichuan, China

Int. J. Mol. Sci. 2012, 13(11), 15279-15290; https://doi.org/10.3390/ijms131115279 - 19 Nov 2012

Cited by 51 | Viewed by 10313

Abstract

Ionic self-assembly of the peptide RADARADARADARADA (RADA16-1) may form a well-defined nanofiber and eventually a hydrogel scaffold, with a water content of over 99.5%. This leads to the establishment of a nanofiber barrier that can be used to achieve complete hemostasis in less [...] Read more.

Ionic self-assembly of the peptide RADARADARADARADA (RADA16-1) may form a well-defined nanofiber and eventually a hydrogel scaffold, with a water content of over 99.5%. This leads to the establishment of a nanofiber barrier that can be used to achieve complete hemostasis in less than 20 s in multiple tissues and in a variety of different wounds. In the present study, the nanofiber scaffolds of RADA16-1 peptide were sonicated into smaller fragments to identify possible molecular mechanisms underlying the rapid cessation of bleeding associated with these materials. Atomic force microscopy (AFM), circular dichroism (CD), and rheometry were also used to evaluate the re-assembly kinetics of this peptide. A bleeding control experiment was performed in animal models to uncover the molecular mechanisms underlying this fast hemostasis. In this way, these sonicated fragments not only quickly reassembled into nanofibers indistinguishable from the original material, but the degree of reassembly was also correlated with an increase in the rigidity of the scaffold and increased as the time required for hemostasis increased. Full article

(This article belongs to the Section Biochemistry)

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14 pages, 3436 KiB

Open AccessArticle

Clinicopathological Signiﬁcance of NMIIA Overexpression in Human Gastric Cancer

by Dongning Liu^†, Lei Zhang^†, Zhiyong Shen, Fei Tan, Yanfeng Hu, Jiang Yu and Guoxin Li^*

¹ Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 15291-15304; https://doi.org/10.3390/ijms131115291 - 19 Nov 2012

Cited by 34 | Viewed by 8479

Abstract

Altered expressions of nonmuscle myosin IIA (NMIIA) have been observed in certain types of cancers, but the impact of the alterations in gastric cancer (GC) remains unclear. The purpose of this study was to evaluate the expression of NMIIA at the mRNA and [...] Read more.

Altered expressions of nonmuscle myosin IIA (NMIIA) have been observed in certain types of cancers, but the impact of the alterations in gastric cancer (GC) remains unclear. The purpose of this study was to evaluate the expression of NMIIA at the mRNA and protein level in patients with GC and to assess its clinical significance. We investigated the expression of NMIIA in fresh, paired GC tissues by reverse transcriptase polymerase chain reaction (RT-PCR; n = 14) and Western blot analysis (n = 36). Simultaneously, we performed immunohistochemistry (IHC) on paraffin embedded specimens, including 96 GC specimens, 30 matched normal specimens and 30 paired metastatic lymph node samples. NMIIA is overexpressed in GC compared with the adjacent normal gastric epithelium (p < 0.001) and high-level NMIIA expression is significantly correlated with the depth of wall invasion, lymph node metastasis, distant metastasis and Tumor Node Metastasis (TNM) stage. Furthermore, elevated NMIIA expression is an independent prognostic factor in multivariate analysis using the Cox regression model (p = 0.021). These findings indicate that overexpression of NMIIA may contribute to the progression and poor prognosis of GC. Full article

(This article belongs to the Special Issue Advances in Cancer Diagnosis)

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16 pages, 761 KiB

Open AccessArticle

Pharmacological Evaluation and Preliminary Pharmacokinetics Studies of a New Diclofenac Prodrug without Gastric Ulceration Effect

by Jean Leandro dos Santos^1,*, Vanessa Moreira², Michel Leandro Campos³, Rafael Consolin Chelucci¹, Karina Pereira Barbieri¹, Pollyana Cristina Maggio de Castro Souto², Márcio Hideki Matsubara², Catarina Teixeira², Priscila Longhin Bosquesi¹, Rosângela Gonçalves Peccinini³ and Chung Man Chin^1,*

¹ Lapdesf—Laboratory of Research and Drug Development, Drugs and Medicines Department, School of Pharmaceutical Science—UNESP Rodovia Araraquara Jaú Km. 01, 14801-902, Araraquara, SP, Brazil

² Pharmacology Laboratory, Inflammation unit, Butantan Institute, Vital Brazil Avenue 1500, 05503-900, São Paulo, Brazil

³ Natural Active Principles and Toxicology Department, School of Pharmaceutical Science—UNESP Rodovia Araraquara Jaú Km. 01, 14801-902, Araraquara, SP, Brazil

Int. J. Mol. Sci. 2012, 13(11), 15305-15320; https://doi.org/10.3390/ijms131115305 - 19 Nov 2012

Cited by 13 | Viewed by 7992

Abstract

Long-term nonsteroidal anti-inflammatory drugs (NSAIDs) therapy has been associated with several adverse effects such as gastric ulceration and cardiovascular events. Among the molecular modifications strategies, the prodrug approach is a useful tool to discover new safe NSAIDs. The 1-(2,6-dichlorophenyl)indolin-2-one is a diclofenac prodrug [...] Read more.

Long-term nonsteroidal anti-inflammatory drugs (NSAIDs) therapy has been associated with several adverse effects such as gastric ulceration and cardiovascular events. Among the molecular modifications strategies, the prodrug approach is a useful tool to discover new safe NSAIDs. The 1-(2,6-dichlorophenyl)indolin-2-one is a diclofenac prodrug which demonstrated relevant anti-inflammatory properties without gastro ulceration effect. In addition, the prodrug decreases PGE₂ levels, COX-2 expression and cellular influx into peritoneal cavity induced by carrageenan treatment. Preliminary pharmacokinetic studies have shown in vivo bioconversion of prodrug to diclofenac. This prodrug is a new nonulcerogenic NSAID useful to treat inflammatory events by long-term therapy. Full article

(This article belongs to the Section Biochemistry)

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Graphical abstract

22 pages, 266 KiB

Open AccessArticle

Primary, Secondary Metabolites, Photosynthetic Capacity and Antioxidant Activity of the Malaysian Herb Kacip Fatimah (Labisia Pumila Benth) Exposed to Potassium Fertilization under Greenhouse Conditions

by Mohd Hafiz Ibrahim, Hawa Z. E. Jaafar^*, Ehsan Karimi

and Ali Ghasemzadeh

Department of Crop Science, Faculty of Agriculture, University Putra Malaysia, 43400 Serdang, Selangor, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 15321-15342; https://doi.org/10.3390/ijms131115321 - 20 Nov 2012

Cited by 94 | Viewed by 9336

Abstract

A randomized complete block design was used to characterize the relationship between production of total phenolics, flavonoids, ascorbic acid, carbohydrate content, leaf gas exchange, phenylalanine ammonia-lyase (PAL), soluble protein, invertase and antioxidant enzyme activities (ascorbate peroxidase (APX), catalase (CAT) and superoxide dismutase (SOD) [...] Read more.

A randomized complete block design was used to characterize the relationship between production of total phenolics, flavonoids, ascorbic acid, carbohydrate content, leaf gas exchange, phenylalanine ammonia-lyase (PAL), soluble protein, invertase and antioxidant enzyme activities (ascorbate peroxidase (APX), catalase (CAT) and superoxide dismutase (SOD) in Labisia pumila Benth var. alata under four levels of potassium fertilization experiments (0, 90, 180 and 270 kg K/ha) conducted for 12 weeks. It was found that the production of total phenolics, flavonoids, ascorbic acid and carbohydrate content was affected by the interaction between potassium fertilization and plant parts. As the potassium fertilization levels increased from 0 to 270 kg K/ha, the production of soluble protein and PAL activity increased steadily. At the highest potassium fertilization (270 kg K/ha) L. pumila exhibited signiﬁcantly higher net photosynthesis (A), stomatal conductance (g_s), intercellular CO₂ (C_i), apparent quantum yield (ɸ) and lower dark respiration rates (R_d), compared to the other treatments. It was found that the production of total phenolics, flavonoids and ascorbic acid are also higher under 270 kg K/ha compared to 180, 90 and 0 kg K/ha. Furthermore, from the present study, the invertase activity was also found to be higher in 270 kg K/ha treatment. The antioxidant enzyme activities (APX, CAT and SOD) were lower under high potassium fertilization (270 kg K/ha) and have a significant negative correlation with total phenolics and flavonoid production. From this study, it was observed that the up-regulation of leaf gas exchange and downregulation of APX, CAT and SOD activities under high supplementation of potassium fertilizer enhanced the carbohydrate content that simultaneously increased the production of L. pumila secondary metabolites, thus increasing the health promoting effects of this plant. Full article

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17 pages, 453 KiB

Open AccessArticle

Targeting Death Receptor TRAIL-R2 by Chalcones for TRAIL-Induced Apoptosis in Cancer Cells

by Ewelina Szliszka^†, Dagmara Jaworska^†

, Małgorzata Ksek, Zenon P. Czuba and Wojciech Król^*

¹ Chair and Department of Microbiology and Immunology, Medical University of Silesia in Katowice, Jordana 19, 41-808 Zabrze, Poland

^† These authors contributed equally to this work.

Int. J. Mol. Sci. 2012, 13(11), 15343-15359; https://doi.org/10.3390/ijms131115343 - 20 Nov 2012

Cited by 58 | Viewed by 8900

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in cancer cells without toxicity to normal cells. TRAIL binds to death receptors, TRAIL-R1 (DR4) and TRAIL-R2 (DR5) expressed on cancer cell surface and activates apoptotic pathways. Endogenous TRAIL plays an important role in immune [...] Read more.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in cancer cells without toxicity to normal cells. TRAIL binds to death receptors, TRAIL-R1 (DR4) and TRAIL-R2 (DR5) expressed on cancer cell surface and activates apoptotic pathways. Endogenous TRAIL plays an important role in immune surveillance and defense against cancer cells. However, as more tumor cells are reported to be resistant to TRAIL mediated death, it is important to search for and develop new strategies to overcome this resistance. Chalcones can sensitize cancer cells to TRAIL-induced apoptosis. We examined the cytotoxic and apoptotic effects of TRAIL in combination with four chalcones: chalcone, isobavachalcone, licochalcone A and xanthohumol on HeLa cancer cells. The cytotoxicity was measured by MTT and LDH assays. The apoptosis was detected using annexin V-FITC staining by flow cytometry and fluorescence microscopy. Death receptor expression was analyzed using flow cytometry. The decreased expression of death receptors in cancer cells may be the cause of TRAIL-resistance. Chalcones enhance TRAIL-induced apoptosis in HeLa cells through increased expression of TRAIL-R2. Our study has indicated that chalcones augment the antitumor activity of TRAIL and confirm their cancer chemopreventive properties. Full article

(This article belongs to the Special Issue Advances in Molecular Oncology (special issue))

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13 pages, 547 KiB

Open AccessArticle

Eclipsed Acetaldehyde as a Precursor for Producing Vinyl Alcohol

by Osman I. Osman^*

, Abdulrahman O. Alyoubi, Shabaan A. K. Elroby

, Rifaat H. Hilal and Saadullah G. Aziz

Chemistry Department, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia

Int. J. Mol. Sci. 2012, 13(11), 15360-15372; https://doi.org/10.3390/ijms131115360 - 20 Nov 2012

Cited by 5 | Viewed by 8245

Abstract

The MP2 and DFT/B3LYP methods at 6-311++G(d,p) and aug-cc-pdz basis sets have been used to probe the origin of relative stability preference for eclipsed acetaldehyde over its bisected counterpart. A relative energy stability range of 1.02 to 1.20 kcal/mol, in favor of the [...] Read more.

The MP2 and DFT/B3LYP methods at 6-311++G(d,p) and aug-cc-pdz basis sets have been used to probe the origin of relative stability preference for eclipsed acetaldehyde over its bisected counterpart. A relative energy stability range of 1.02 to 1.20 kcal/mol, in favor of the eclipsed conformer, was found and discussed. An NBO study at these chemistry levels complemented these findings and assigned the eclipsed acetaldehyde preference mainly to the vicinal antiperiplanar hyperconjugative interactions. The tautomeric interconversion between the more stable eclipsed acetaldehyde and vinyl alcohol has been achieved through a four-membered ring transition state (TS). The obtained barrier heights and relative stabilities of eclipsed acetaldehyde and the two conformers of vinyl alchol at these model chemistries have been estimated and discussed. Full article

(This article belongs to the Section Physical Chemistry, Theoretical and Computational Chemistry)

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14 pages, 2170 KiB

Open AccessArticle

Tenomodulin Inhibits Retinal Neovascularization in a Mouse Model of Oxygen-Induced Retinopathy

by Wei Wang^1,*, Zhongqiu Li¹, Tatsuhiko Sato² and Yusuke Oshima²

¹ Department of Ophthalmology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China

² Department of Ophthalmology and Visual Science, Osaka University Medical School, Yamadaoka, Suita, Osaka 5650871, Japan

Int. J. Mol. Sci. 2012, 13(11), 15373-15386; https://doi.org/10.3390/ijms131115373 - 20 Nov 2012

Cited by 10 | Viewed by 7155

Abstract

We aimed to determine the anti-angiogenic effect of tenomodulin (TeM) on retinal neovascularization in an oxygen-induced retinopathy (OIR) mouse model. OIR was induced in C57BL/6 mice by exposing seven-day-old mice to 75% oxygen for five days followed by room air for five days. [...] Read more.

We aimed to determine the anti-angiogenic effect of tenomodulin (TeM) on retinal neovascularization in an oxygen-induced retinopathy (OIR) mouse model. OIR was induced in C57BL/6 mice by exposing seven-day-old mice to 75% oxygen for five days followed by room air for five days. Control mice were exposed to room air from birth until postnatal day 17. Mice received intravitreal injections of 1 μg of TeM in one eye and PBS in the contralateral eye at P7 before being exposed to 75% oxygen. Eyes were collected at postnatal day 17. Retinal blood vessel patterns were visualized by fluorescein angiography. We quantified the number of neovascular nuclei that were present beyond the inner limiting membrane (ILM) using histological methods with a masked approach. Furthermore, double immunohistochemical staining of TeM was performed on retinas to identify nuclei protruding into the vitreous cavity. Western blot was used to detect exogenous TeM protein. The central nonperfusion area (NPA, mm²) of TeM-injected eyes was significantly different from that of OIR and PBS-injected eyes, and the number of nuclei in new blood vessels breaking through the ILM in each retinal cross-section significantly differed from that of OIR eyes and PBS-injected control eyes. Cellular nuclei of new blood vessels protruding into the vitreous cavity were also observed in TeM-injected retinas by immunohistochemistry. Western blotting revealed a 16-kDa immunoreactive protein, indicating incorporation of an exogenous TeM fragment into the retina. Our data shows that TeM can effectively inhibit pathological angiogenesis in mouse eyes; indicating its potential role in prevention and treatment of ocular neovascularization. Full article

(This article belongs to the Section Biochemistry)

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14 pages, 414 KiB

Open AccessArticle

Predicting Retention Times of Naturally Occurring Phenolic Compounds in Reversed-Phase Liquid Chromatography: A Quantitative Structure-Retention Relationship (QSRR) Approach

by Jamshed Akbar¹, Shahid Iqbal¹, Fozia Batool¹, Abdul Karim¹ and Kim Wei Chan^2,*

¹ Department of Chemistry, University of Sargodha, Sargodha 40100, Pakistan

² Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Serdang 43400, Malaysia

Int. J. Mol. Sci. 2012, 13(11), 15387-15400; https://doi.org/10.3390/ijms131115387 - 20 Nov 2012

Cited by 22 | Viewed by 7516

Abstract

Quantitative structure-retention relationships (QSRRs) have successfully been developed for naturally occurring phenolic compounds in a reversed-phase liquid chromatographic (RPLC) system. A total of 1519 descriptors were calculated from the optimized structures of the molecules using MOPAC2009 and DRAGON softwares. The data set of [...] Read more.

Quantitative structure-retention relationships (QSRRs) have successfully been developed for naturally occurring phenolic compounds in a reversed-phase liquid chromatographic (RPLC) system. A total of 1519 descriptors were calculated from the optimized structures of the molecules using MOPAC2009 and DRAGON softwares. The data set of 39 molecules was divided into training and external validation sets. For feature selection and mapping we used step-wise multiple linear regression (SMLR), unsupervised forward selection followed by step-wise multiple linear regression (UFS-SMLR) and artificial neural networks (ANN). Stable and robust models with significant predictive abilities in terms of validation statistics were obtained with negation of any chance correlation. ANN models were found better than remaining two approaches. HNar, IDM, Mp, GATS2v, DISP and 3D-MoRSE (signals 22, 28 and 32) descriptors based on van der Waals volume, electronegativity, mass and polarizability, at atomic level, were found to have significant effects on the retention times. The possible implications of these descriptors in RPLC have been discussed. All the models are proven to be quite able to predict the retention times of phenolic compounds and have shown remarkable validation, robustness, stability and predictive performance. Full article

(This article belongs to the Section Biochemistry)

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19 pages, 585 KiB

Open AccessReview

Marine Omega-3 Phospholipids: Metabolism and Biological Activities

by Lena Burri¹, Nils Hoem¹, Sebastiano Banni² and Kjetil Berge^1,*

¹ Aker BioMarine ASA, Fjordalléen 16, NO-0115 Oslo, Norway

² Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy

Int. J. Mol. Sci. 2012, 13(11), 15401-15419; https://doi.org/10.3390/ijms131115401 - 21 Nov 2012

Cited by 228 | Viewed by 24799

Abstract

The biological activities of omega-3 fatty acids (n-3 FAs) have been under extensive study for several decades. However, not much attention has been paid to differences of dietary forms, such as triglycerides (TGs) versus ethyl esters or phospholipids (PLs). New innovative marine raw [...] Read more.

The biological activities of omega-3 fatty acids (n-3 FAs) have been under extensive study for several decades. However, not much attention has been paid to differences of dietary forms, such as triglycerides (TGs) versus ethyl esters or phospholipids (PLs). New innovative marine raw materials, like krill and fish by-products, present n-3 FAs mainly in the PL form. With their increasing availability, new evidence has emerged on n-3 PL biological activities and differences to n-3 TGs. In this review, we describe the recently discovered nutritional properties of n-3 PLs on different parameters of metabolic syndrome and highlight their different metabolic bioavailability in comparison to other dietary forms of n-3 FAs. Full article

(This article belongs to the Special Issue Phospholipids: Molecular Sciences 2012)

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27 pages, 2386 KiB

Open AccessArticle

Errors in the Calculation of ²⁷Al Nuclear Magnetic Resonance Chemical Shifts

by Xianlong Wang^1,2,*, Chengfei Wang¹ and Hui Zhao¹

¹ Center of Bioinformatics, University of Electronic Science and Technology of China, No. 4, 2nd Section, Jianshe Road, Chengdu 610054, China

² Department of Chemistry, Bryn Mawr College, 101 North Merion Avenue, Bryn Mawr, PA 19010, USA

Int. J. Mol. Sci. 2012, 13(11), 15420-15446; https://doi.org/10.3390/ijms131115420 - 21 Nov 2012

Cited by 9 | Viewed by 8026

Abstract

Computational chemistry is an important tool for signal assignment of ²⁷Al nuclear magnetic resonance spectra in order to elucidate the species of aluminum(III) in aqueous solutions. The accuracy of the popular theoretical models for computing the ²⁷Al chemical shifts was evaluated [...] Read more.

Computational chemistry is an important tool for signal assignment of ²⁷Al nuclear magnetic resonance spectra in order to elucidate the species of aluminum(III) in aqueous solutions. The accuracy of the popular theoretical models for computing the ²⁷Al chemical shifts was evaluated by comparing the calculated and experimental chemical shifts in more than one hundred aluminum(III) complexes. In order to differentiate the error due to the chemical shielding tensor calculation from that due to the inadequacy of the molecular geometry prediction, single-crystal X-ray diffraction determined structures were used to build the isolated molecule models for calculating the chemical shifts. The results were compared with those obtained using the calculated geometries at the B3LYP/6-31G(d) level. The isotropic chemical shielding constants computed at different levels have strong linear correlations even though the absolute values differ in tens of ppm. The root-mean-square difference between the experimental chemical shifts and the calculated values is approximately 5 ppm for the calculations based on the X-ray structures, but more than 10 ppm for the calculations based on the computed geometries. The result indicates that the popular theoretical models are adequate in calculating the chemical shifts while an accurate molecular geometry is more critical. Full article

(This article belongs to the Special Issue Atoms in Molecules and in Nanostructures)

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Graphical abstract

17 pages, 189 KiB

Open AccessReview

Dietary Docosahexaenoic Acid (22:6) Incorporates into Cardiolipin at the Expense of Linoleic Acid (18:2): Analysis and Potential Implications

by Colin H. Cortie^* and Paul L. Else

Metabolic Research Centre (in IHMRI), School of Health Sciences, University of Wollongong, Wollongong, NSW 2522, Australia

Int. J. Mol. Sci. 2012, 13(11), 15447-15463; https://doi.org/10.3390/ijms131115447 - 21 Nov 2012

Cited by 26 | Viewed by 9389

Abstract

Cardiolipin is a signature phospholipid of major functional significance in mitochondria. In heart mitochondria the fatty acid composition of cardiolipin is commonly viewed as highly regulated due to its high levels of linoleic acid (18:2n − 6) and the dominant presence of [...] Read more.

Cardiolipin is a signature phospholipid of major functional significance in mitochondria. In heart mitochondria the fatty acid composition of cardiolipin is commonly viewed as highly regulated due to its high levels of linoleic acid (18:2n − 6) and the dominant presence of a 4×18:2 molecular species. However, analysis of data from a comprehensive compilation of studies reporting changes in fatty acid composition of cardiolipin in heart and liver mitochondria in response to dietary fat shows that, in heart the accrual of 18:2 into cardiolipin conforms strongly to its dietary availability at up to 20% of total dietary fatty acid and thereafter is regulated. In liver, no dietary conformer trend is apparent for 18:2 with regulated lower levels across the dietary range for 18:2. When 18:2 and docosahexaenoic acid (22:6n − 3) are present in the same diet, 22:6 is incorporated into cardiolipin of heart and liver at the expense of 18:2 when 22:6 is up to ~20% and 10% of total dietary fatty acid respectively. Changes in fatty acid composition in response to dietary fat are also compared for the two other main mitochondrial phospholipids, phosphatidylcholine and phosphatidylethanolamine, and the potential consequences of replacement of 18:2 with 22:6 in cardiolipin are discussed. Full article

(This article belongs to the Special Issue Phospholipids: Molecular Sciences 2012)

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11 pages, 219 KiB

Open AccessArticle

Associations between Endogenous Dimethylarginines and Renal Function in Healthy Children and Adolescents

by Aleksandra Jaźwińska-Kozuba¹, Jens Martens-Lobenhoffer², Andrzej Surdacki^3,*, Olga Kruszelnicka⁴, Jarosław Rycaj⁵, Urszula Godula-Stuglik⁶ and Stefanie M. Bode-Böger²

¹ Almed-Elektra Medical Center, 41-710 Ruda Śląska, Poland

² Institute for Clinical Pharmacology, Otto-von-Guericke University, 39120 Magdeburg, Germany

³ Department of Cardiology, Jagiellonian University/University Hospital, 31-501 Cracow, Poland

⁴ Department of Coronary Artery Disease, the John Paul II Hospital, 31-202 Cracow, Poland

⁵ Department of Cardiology, Congenital Heart Defects and Electrotherapy, Silesian Center for Heart Diseases in Zabrze, 41-800 Zabrze, Medical University of Silesia, Poland

⁶ Department of Pediatrics in Zabrze, 41-800 Zabrze, Medical University of Silesia, Poland

Int. J. Mol. Sci. 2012, 13(11), 15464-15474; https://doi.org/10.3390/ijms131115464 - 21 Nov 2012

Cited by 6 | Viewed by 6130

Abstract

The structural isomer of asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), is eliminated almost entirely by urinary excretion and considered a sensitive index of glomerular filtration rate (GFR). However, reports on this relationship in healthy subjects younger than 18 years of age are rare. [...] Read more.

The structural isomer of asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), is eliminated almost entirely by urinary excretion and considered a sensitive index of glomerular filtration rate (GFR). However, reports on this relationship in healthy subjects younger than 18 years of age are rare. Therefore, our aim was to investigate relations between endogenous dimethylarginines and renal function indices in healthy children and adolescents. We studied 40 subjects aged 3–18 years free of coexistent diseases or subclinical carotid atherosclerosis. A serum creatinine-derived estimated GFR (eGFR) was calculated by the revised bedside Schwartz equation. L-arginine, ADMA and SDMA were measured by liquid chromatography-tandem mass spectrometry. Mean eGFR was 122 ± 22 (SD) mL/min per 1.73 m². Creatinine and eGFR exhibited closer correlations with the SDMA/ADMA ratio (r = 0.64, p < 0.0001; r = −0.63, p < 0.0001, respectively) than with SDMA (r = 0.31, p = 0.05; r = −0.35, p = 0.03). Neither creatinine nor eGFR correlated with ADMA or L-arginine. Adjustment for age or height only slightly attenuated the associations between the SDMA/ADMA ratio and eGFR or creatinine. Our findings suggest the superiority of the SDMA/ADMA ratio over SDMA as a renal function index in healthy children. Thus, further studies are warranted to verify our preliminary results in a larger group of subjects below 18 years of age. Full article

(This article belongs to the Special Issue ADMA and Nitrergic System)

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Int. J. Mol. Sci., Volume 13, Issue 11 (November 2012) – 105 articles , Pages 13764-15495

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