19 pages, 3262 KiB  
Article
Increased Secreted Frizzled-Related Protein 5 mRNA Expression in the Adipose Tissue of Women with Nonalcoholic Fatty Liver Disease Associated with Obesity
by Laia Bertran 1,†, Marta Portillo-Carrasquer 1,†, Andrea Barrientos-Riosalido 1, Carmen Aguilar 1, David Riesco 2, Salomé Martínez 3, Amada Culebradas 4, Margarita Vives 4, Fàtima Sabench 4, Daniel Del Castillo 4, Cristóbal Richart 1,2 and Teresa Auguet 1,2,*
1 Grup de Recerca GEMMAIR (AGAUR)—Medicina Aplicada (URV), Departament de Medicina i Cirurgia, Universitat Rovira i Virgili (URV), Institut d’Investigació Sanitària Pere Virgili (IISPV), 43007 Tarragona, Spain
2 Servei Medicina Interna, Hospital Universitari de Tarragona Joan XXIII, Mallafré Guasch, 4, 43007 Tarragona, Spain
3 Servei Anatomia Patològica, Hospital Universitari de Tarragona Joan XXIII, Mallafré Guasch, 4, 43007 Tarragona, Spain
4 Servei de Cirurgia, Hospital Sant Joan de Reus, Departament de Medicina i Cirurgia, Universitat Rovira i Virgili (URV), IISPV, Avinguda Doctor Josep Laporte, 2, 43204 Reus, Spain
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(17), 9871; https://doi.org/10.3390/ijms23179871 - 30 Aug 2022
Cited by 2 | Viewed by 2956
Abstract
Secreted frizzled-related protein 5 (SFRP5) is an anti-inflammatory adipocytokine secreted by adipocytes that seems to be linked with nonalcoholic fatty liver disease (NAFLD). We aimed to evaluate the role of the SFRP5-wingless-MMTV integration site family member 5a (WNT5A) pathway, closely related to adipogenesis, [...] Read more.
Secreted frizzled-related protein 5 (SFRP5) is an anti-inflammatory adipocytokine secreted by adipocytes that seems to be linked with nonalcoholic fatty liver disease (NAFLD). We aimed to evaluate the role of the SFRP5-wingless-MMTV integration site family member 5a (WNT5A) pathway, closely related to adipogenesis, in subcutaneous (SAT) and visceral adipose tissues (VAT) and its relationship with obesity-related NAFLD. Our cohort was composed of 60 women with morbid obesity (MO), who underwent hypocaloric diet, subclassified according to their hepatic histopathology and 15 women with normal weight. We observed increased SFRP5 mRNA expression in VAT and lower WNT5A expression in SAT in MO compared to normal weight. We found elevated SFRP5 expression in nonalcoholic steatohepatitis (NASH) in SAT and in mild simple steatosis (SS) and NASH in VAT. We observed higher WNT5A expression in SS compared to normal liver in SAT, and a peak of WNT5A expression in mild SS. To conclude, we reported increased SFRP5 mRNA expression in SAT and VAT of NAFLD-related to obesity subjects, suggesting an implication of the SFRP5-WNT5A pathway in NAFLD pathogenesis, probably due to the adipose tissue-liver axis. Since the mechanisms by which this potential interaction takes place remain elusive, more research in this field is needed. Full article
(This article belongs to the Special Issue Nutrition and Metabolism in Health and Disease: From Gene to Organism)
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15 pages, 2239 KiB  
Article
Preliminary Characterization of the Epigenetic Modulation in the Human Mesenchymal Stem Cells during Chondrogenic Process
by Marco Miceli 1,2,*, Giuseppe Maria Maruotti 1, Laura Sarno 1, Luigi Carbone 1, Maurizio Guida 1,† and Alessandra Pelagalli 3,4,*,†
1 Department of Neuroscience, Reproductive and Odontostomatological Sciences, University of Naples “Federico II”, 80131 Naples, Italy
2 CEINGE Biotecnologie Avanzate, 80145 Naples, Italy
3 Department of Advanced Biomedical Sciences, University of Naples “Federico II”, 80131 Naples, Italy
4 Institute of Biostructures and Bioimaging (IBB), National Research Council (CNR), 80131 Naples, Italy
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(17), 9870; https://doi.org/10.3390/ijms23179870 - 30 Aug 2022
Cited by 1 | Viewed by 1701
Abstract
Regenerative medicine represents a growing hot topic in biomedical sciences, aiming at setting out novel therapeutic strategies to repair or regenerate damaged tissues and organs. For this perspective, human mesenchymal stem cells (hMSCs) play a key role in tissue regeneration, having the potential [...] Read more.
Regenerative medicine represents a growing hot topic in biomedical sciences, aiming at setting out novel therapeutic strategies to repair or regenerate damaged tissues and organs. For this perspective, human mesenchymal stem cells (hMSCs) play a key role in tissue regeneration, having the potential to differentiate into many cell types, including chondrocytes. Accordingly, in the last few years, researchers have focused on several in vitro strategies to optimize hMSC differentiation protocols, including those relying on epigenetic manipulations that, in turn, lead to the modulation of gene expression patterns. Therefore, in the present study, we investigated the role of the class II histone deacetylase (HDAC) inhibitor, MC1568, in the hMSCs-derived chondrogenesis. The hMSCs we used for this work were the hMSCs obtained from the amniotic fluid, given their greater differentiation capacity. Our preliminary data documented that MC1568 drove both the improvement and acceleration of hMSCs chondrogenic differentiation in vitro, since the differentiation process in MC1568-treated cells took place in about seven days, much less than that normally observed, namely 21 days. Collectively, these preliminary data might shed light on the validity of such a new differentiative protocol, in order to better assess the potential role of the epigenetic modulation in the process of the hypertrophic cartilage formation, which represents the starting point for endochondral ossification. Full article
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22 pages, 4061 KiB  
Article
Analysis of Dormancy-Associated Transcriptional Networks Reveals a Shared Quiescence Signature in Lung and Colorectal Cancer
by Adriano Cuccu 1, Federica Francescangeli 1, Maria Laura De Angelis 1, Alessandro Bruselles 1, Alessandro Giuliani 2 and Ann Zeuner 1,*
1 Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
2 Environment and Health Department, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy
Int. J. Mol. Sci. 2022, 23(17), 9869; https://doi.org/10.3390/ijms23179869 - 30 Aug 2022
Cited by 17 | Viewed by 3465
Abstract
Quiescent cancer cells (QCCs) are a common feature of solid tumors, representing a major obstacle to the long-term success of cancer therapies. We isolated QCCs ex vivo from non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) xenografts with a label-retaining strategy and [...] Read more.
Quiescent cancer cells (QCCs) are a common feature of solid tumors, representing a major obstacle to the long-term success of cancer therapies. We isolated QCCs ex vivo from non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) xenografts with a label-retaining strategy and compared QCCs gene expression profiles to identify a shared “quiescence signature”. Principal Component Analysis (PCA) revealed a specific component neatly discriminating quiescent and replicative phenotypes in NSCLC and CRC. The discriminating component showed significant overlapping, with 688 genes in common including ZEB2, a master regulator of stem cell plasticity and epithelial-to-mesenchymal transition (EMT). Gene set enrichment analysis showed that QCCs of both NSCLC and CRC had an increased expression of factors related to stemness/self renewal, EMT, TGF-β, morphogenesis, cell adhesion and chemotaxis, whereas proliferating cells overexpressed Myc targets and factors involved in RNA metabolism. Eventually, we analyzed in depth by means of a complex network approach, both the ‘morphogenesis module’ and the subset of differentially expressed genes shared by NCSLC and CRC. This allowed us to recognize different gene regulation network wiring for quiescent and proliferating cells and to underpin few genes central for network integration that may represent new therapeutic vulnerabilities. Altogether, our results highlight common regulatory pathways in QCCs of lung and colorectal tumors that may be the target of future therapeutic interventions. Full article
(This article belongs to the Special Issue Advances in Genome Regulation in Cancer)
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24 pages, 8139 KiB  
Review
An Eye on Kupffer Cells: Development, Phenotype and the Macrophage Niche
by Andrey Elchaninov 1,2,*, Polina Vishnyakova 1,3, Egor Menyailo 4, Gennady Sukhikh 1 and Timur Fatkhudinov 3,4
1 Laboratory of Regenerative Medicine, National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of Ministry of Healthcare of Russian Federation, 117997 Moscow, Russia
2 Histology Department, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
3 Histology Department, Medical Institute, Peoples’ Friendship University of Russia, 117198 Moscow, Russia
4 Laboratory of Growth and Development, Avtsyn Research Institute of Human Morphology of FSBI “Petrovsky National Research Centre of Surgery”, 117418 Moscow, Russia
Int. J. Mol. Sci. 2022, 23(17), 9868; https://doi.org/10.3390/ijms23179868 - 30 Aug 2022
Cited by 26 | Viewed by 8952
Abstract
Macrophages are key participants in the maintenance of tissue homeostasis under normal and pathological conditions, and implement a rich diversity of functions. The largest population of resident tissue macrophages is found in the liver. Hepatic macrophages, termed Kupffer cells, are involved in the [...] Read more.
Macrophages are key participants in the maintenance of tissue homeostasis under normal and pathological conditions, and implement a rich diversity of functions. The largest population of resident tissue macrophages is found in the liver. Hepatic macrophages, termed Kupffer cells, are involved in the regulation of multiple liver functionalities. Specific differentiation profiles and functional activities of tissue macrophages have been attributed to the shaping role of the so-called tissue niche microenvironments. The fundamental macrophage niche concept was lately shaken by a flood of new data, leading to a revision and substantial update of the concept, which constitutes the main focus of this review. The macrophage community discusses contemporary evidence on the developmental origins of resident macrophages, notably Kupffer cells and the issues of heterogeneity of the hepatic macrophage populations, as well as the roles of proliferation, cell death and migration processes in the maintenance of macrophage populations of the liver. Special consideration is given to interactions of Kupffer cells with other local cell lineages, including Ito cells, sinusoidal endothelium and hepatocytes, which participate in the maintenance of their phenotypical and functional identity. Full article
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13 pages, 1774 KiB  
Article
Absorption and Metabolism of the Natural Sweeteners Erythritol and Xylitol in Humans: A Dose-Ranging Study
by Valentine Bordier 1,2, Fabienne Teysseire 1,2, Frank Senner 3, Götz Schlotterbeck 3, Jürgen Drewe 4, Christoph Beglinger 1,2, Bettina K. Wölnerhanssen 1,2,*,† and Anne Christin Meyer-Gerspach 1,2,*,†
1 St. Clara Research Ltd., St. Claraspital, 4002 Basel, Switzerland
2 Faculty of Medicine, University of Basel, 4001 Basel, Switzerland
3 Institute for Chemistry and Bioanalytics, School of Life Science, FHNW University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland
4 Department of Clinical Pharmacology and Toxicology, University Hospital Basel, 4001 Basel, Switzerland
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(17), 9867; https://doi.org/10.3390/ijms23179867 - 30 Aug 2022
Cited by 20 | Viewed by 6843
Abstract
The natural sweeteners erythritol and xylitol might be helpful to reduce sugar consumption and therefore prevent obesity and diabetes. The aim of the present study was to determine the absorption and metabolization into erythronate of different concentrations of erythritol and xylitol. Seventeen healthy [...] Read more.
The natural sweeteners erythritol and xylitol might be helpful to reduce sugar consumption and therefore prevent obesity and diabetes. The aim of the present study was to determine the absorption and metabolization into erythronate of different concentrations of erythritol and xylitol. Seventeen healthy lean participants received intragastric solutions of 10, 25, or 50 g erythritol or 7, 17, or 35 g xylitol on three study days in a randomized order. The study was double blinded with respect to the doses administered. We assessed plasma concentrations of erythritol, xylitol, and erythronate at fixed time intervals after administration with gas chromatography-mass spectrometry. We found: (i) a dose-dependent and saturable absorption of erythritol, (ii) a very low absorption of xylitol, (iii) a dose-dependent metabolization of erythritol into erythronate, and (iv) no metabolization of xylitol into erythronate. The implications of the metabolization of erythritol into erythronate for human health remain to be determined and more research in this area is needed. Full article
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24 pages, 3782 KiB  
Review
The Mechanism-Based Inactivation of CYP3A4 by Ritonavir: What Mechanism?
by Nancy H. C. Loos 1, Jos H. Beijnen 2,3 and Alfred H. Schinkel 1,*
1 The Netherlands Cancer Institute, Division of Pharmacology, 1066 CX Amsterdam, The Netherlands
2 Faculty of Science, Department of Pharmaceutical Sciences, Division of Pharmacoepidemiology and Clinical Pharmacology, Utrecht University, 3584 CS Utrecht, The Netherlands
3 The Netherlands Cancer Institute, Division of Pharmacy and Pharmacology, 1066 CX Amsterdam, The Netherlands
Int. J. Mol. Sci. 2022, 23(17), 9866; https://doi.org/10.3390/ijms23179866 - 30 Aug 2022
Cited by 76 | Viewed by 9422
Abstract
Ritonavir is the most potent cytochrome P450 (CYP) 3A4 inhibitor in clinical use and is often applied as a booster for drugs with low oral bioavailability due to CYP3A4-mediated biotransformation, as in the treatment of HIV (e.g., lopinavir/ritonavir) and more recently COVID-19 (Paxlovid [...] Read more.
Ritonavir is the most potent cytochrome P450 (CYP) 3A4 inhibitor in clinical use and is often applied as a booster for drugs with low oral bioavailability due to CYP3A4-mediated biotransformation, as in the treatment of HIV (e.g., lopinavir/ritonavir) and more recently COVID-19 (Paxlovid or nirmatrelvir/ritonavir). Despite its clinical importance, the exact mechanism of ritonavir-mediated CYP3A4 inactivation is still not fully understood. Nonetheless, ritonavir is clearly a potent mechanism-based inactivator, which irreversibly blocks CYP3A4. Here, we discuss four fundamentally different mechanisms proposed for this irreversible inactivation/inhibition, namely the (I) formation of a metabolic-intermediate complex (MIC), tightly coordinating to the heme group; (II) strong ligation of unmodified ritonavir to the heme iron; (III) heme destruction; and (IV) covalent attachment of a reactive ritonavir intermediate to the CYP3A4 apoprotein. Ritonavir further appears to inactivate CYP3A4 and CYP3A5 with similar potency, which is important since ritonavir is applied in patients of all ethnicities. Although it is currently not possible to conclude what the primary mechanism of action in vivo is, it is unlikely that any of the proposed mechanisms are fundamentally wrong. We, therefore, propose that ritonavir markedly inactivates CYP3A through a mixed set of mechanisms. This functional redundancy may well contribute to its overall inhibitory efficacy. Full article
(This article belongs to the Section Molecular Pharmacology)
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14 pages, 4750 KiB  
Article
Selenoprotein P Concentrations in the Cerebrospinal Fluid and Serum of Individuals Affected by Amyotrophic Lateral Sclerosis, Mild Cognitive Impairment and Alzheimer’s Dementia
by Teresa Urbano 1, Marco Vinceti 1,2,3,*, Jessica Mandrioli 2,4, Annalisa Chiari 4, Tommaso Filippini 1,2,5, Roberta Bedin 2,4, Manuela Tondelli 2,4, Cecilia Simonini 2,4, Giovanna Zamboni 2,4, Misaki Shimizu 6 and Yoshiro Saito 6
1 CREAGEN—Environmental, Genetic and Nutritional Epidemiology Research Center, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 287 Via Campi, 41125 Modena, Italy
2 Center for Neurosciences and Neurotechnology, Department of Biomedical, Metabolic, and Neural Sciences, University of Modena and Reggio Emilia, 287 Via Campi, 41125 Modena, Italy
3 Department of Epidemiology, Boston University School of Public Health, 715 Albany Street, Boston, MA 02118, USA
4 Neurology Unit, Azienda Ospedaliero Universitaria di Modena, 71 Via del Pozzo, 41121 Modena, Italy
5 School of Public Health, University of California Berkeley, 1995 University Avenue, Berkeley, CA 94704, USA
6 Laboratory of Molecular Biology and Metabolism, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan
Int. J. Mol. Sci. 2022, 23(17), 9865; https://doi.org/10.3390/ijms23179865 - 30 Aug 2022
Cited by 23 | Viewed by 3747
Abstract
Selenoprotein P, a selenium-transporter protein, has been hypothesized to play a role in the etiology of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and Alzheimer’s dementia (AD). However, data in humans are scarce and largely confined to autoptic samples. In this case–control [...] Read more.
Selenoprotein P, a selenium-transporter protein, has been hypothesized to play a role in the etiology of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and Alzheimer’s dementia (AD). However, data in humans are scarce and largely confined to autoptic samples. In this case–control study, we determined selenoprotein P concentrations in both the cerebrospinal fluid (CSF) and the serum of 50 individuals diagnosed with ALS, 30 with AD, 54 with mild cognitive impairment (MCI) and of 30 controls, using sandwich enzyme-linked immunosorbent assay (ELISA) methods. We found a positive and generally linear association between CSF and serum selenoprotein P concentrations in all groups. CSF selenoprotein P and biomarkers of neurodegeneration were positively associated in AD, while for MCI, we found an inverted-U-shaped relation. CSF selenoprotein P concentrations were higher in AD and MCI than in ALS and controls, while in serum, the highest concentrations were found in MCI and ALS. Logistic and cubic spline regression analyses showed an inverse association between CSF selenoprotein P levels and ALS risk, and a positive association for AD risk, while an inverted-U-shaped relation with MCI risk emerged. Conversely, serum selenoprotein P concentrations were positively associated with risk of all conditions but only in their lower range. Overall, these findings indicate some abnormalities of selenoprotein P concentrations in both the central nervous system and blood associated with ALS and neurocognitive disorders, though in different directions. These alterations may reflect either phenomena of etiologic relevance or disease-induced alterations of nutritional and metabolic status. Full article
(This article belongs to the Special Issue Molecular Research on Amyotrophic Lateral Sclerosis)
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18 pages, 2668 KiB  
Article
Alpha-Synuclein: The Spark That Flames Dopaminergic Neurons, In Vitro and In Vivo Evidence
by Alexandre Henriques 1,2, Laura Rouvière 1, Elodie Giorla 2, Clémence Farrugia 2, Bilal El Waly 2, Philippe Poindron 1,2 and Noëlle Callizot 1,2,*
1 Neuro-Sys, In Vitro Pharmacology Department, 13120 Gardanne, France
2 Neuro-Sys Vivo, 13120 Gardanne, France
Int. J. Mol. Sci. 2022, 23(17), 9864; https://doi.org/10.3390/ijms23179864 - 30 Aug 2022
Cited by 18 | Viewed by 4034
Abstract
Mitochondria, α-syn fibrils and the endo-lysosomal system are key players in the pathophysiology of Parkinson’s disease. The toxicity of α-syn is amplified by cell-to-cell transmission and aggregation of endogenous species in newly invaded neurons. Toxicity of α-syn PFF was investigated using primary cultures [...] Read more.
Mitochondria, α-syn fibrils and the endo-lysosomal system are key players in the pathophysiology of Parkinson’s disease. The toxicity of α-syn is amplified by cell-to-cell transmission and aggregation of endogenous species in newly invaded neurons. Toxicity of α-syn PFF was investigated using primary cultures of dopaminergic neurons or on aged mice after infusion in the SNpc and combined with mild inhibition of GBA. In primary dopaminergic neurons, application of α-syn PFF induced a progressive cytotoxicity associated with mitochondrial dysfunction, oxidative stress, and accumulation of lysosomes suggesting that exogenous α-syn reached the lysosome (from the endosome). Counteracting the α-syn endocytosis with a clathrin inhibitor, dopaminergic neuron degeneration was prevented. In vivo, α-syn PFF induced progressive neurodegeneration of dopaminergic neurons associated with motor deficits. Histology revealed progressive aggregation of α-syn and microglial activation and accounted for the seeding role of α-syn, injection of which acted as a spark suggesting a triggering of cell-to-cell toxicity. We showed for the first time that a localized SNpc α-syn administration combined with a slight lysosomal deficiency and aging triggered a progressive lesion. The cellular and animal models described could help in the understanding of the human disease and might contribute to the development of new therapies. Full article
(This article belongs to the Special Issue Pathogenesis of Diseases of the Central Nervous System)
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15 pages, 4240 KiB  
Article
Evaluation of the Anti-Atopic Dermatitis Effects of α-Boswellic Acid on Tnf-α/Ifn-γ-Stimulated HaCat Cells and DNCB-Induced BALB/c Mice
by Ya-Chu Tsai 1,2,†, Hsun-Hao Chang 3,†, Sheng-Chieh Chou 4,5,*, Thomas W. Chu 1,6, Yu-Jou Hsu 7, Chien-Yu Hsiao 8, Yuan-Hsin Lo 9, Nan-Lin Wu 10, Der-Chen Chang 11 and Chi-Feng Hung 2,5,12,*
1 Department of Dermatology, Far Eastern Memorial Hospital, New Taipei City 220, Taiwan
2 School of Medicine, Fu Jen Catholic University, New Taipei City 242, Taiwan
3 Department of Cardiology, Tainan Municipal Hospital (Managed by Show Chwan Medical Care Corporation), Tainan City 701, Taiwan
4 Department of Surgery, Taoyuan Armed Force General Hospital, Taoyuan City 325, Taiwan
5 Department of Surgery, Taoyuan Armed Force General Hospital Hsinchu Branch, Hainchu City 300, Taiwan
6 Eastern Virginia Medical School, Norfolk, VA 23507, USA
7 Graduate Institute of Biomedical and Pharmaceutical Science, Fu Jen Catholic University, New Taipei City 242, Taiwan
8 Research Center for Food and Cosmetic Safety, Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan
9 Department of Dermatology, Fu Jen Catholic University Hospital, Fu Jen Catholic University, New Taipei City 242, Taiwan
10 Department of Dermatology, MacKay Memorial Hospital, Taipei 104, Taiwan
11 Department of Mathematics and Statistics, Department of Computer Science, Georgetown University, Washington, DC 20057, USA
12 School of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan
These authors contributed equally to this work.
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Int. J. Mol. Sci. 2022, 23(17), 9863; https://doi.org/10.3390/ijms23179863 - 30 Aug 2022
Cited by 17 | Viewed by 5151
Abstract
Boswellic acids, triterpenoids derived from the genus Boswellia (Burseraceae), are known for their anti-inflammatory and anti-tumor efficacy. Atopic dermatitis is a chronic, non-infectious inflammatory skin disease. However, the effects of α-boswellic acid on atopic dermatitis have not been studied. Therefore, in this study [...] Read more.
Boswellic acids, triterpenoids derived from the genus Boswellia (Burseraceae), are known for their anti-inflammatory and anti-tumor efficacy. Atopic dermatitis is a chronic, non-infectious inflammatory skin disease. However, the effects of α-boswellic acid on atopic dermatitis have not been studied. Therefore, in this study we examined the expression level of pro-inflammatory cytokines, histopathological analysis, and physiological data from BALB/c mice with atopic-like dermatitis induced by 2,4-dinitrochlorobenzene and TNF-α/IFN-γ-stimulated HaCaT cells to better understand the agent’s anti-atopic dermatitis efficacy. First, we found that α-boswellic reduced the epidermal thickening, mast cell numbers, and dermal infiltration of 2,4-dinitrochlorobenzene-induced atopic-like dermatitis in BALB/c mice. Furthermore, we also found that α-boswellic acid can restore transepidermal water loss and skin reddening in mice. In human keratinocytes inflamed by TNF-α/IFN-γ, α-boswellic acid inhibited MAP kinase activation and showed a reduction in NF-κB nuclear translocation. Finally, α-boswellic acid can reduce the expression level of cytokines (IL-1β, IL-6, and IL-8) following the stimulation of TNF-α/IFN-γ in HaCaT cells. Taken together, our study suggests that α-boswellic acids are a potential component for the development of anti-atopic dermatitis drugs. Full article
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15 pages, 1337 KiB  
Review
Prime Editing: An All-Rounder for Genome Editing
by Chenyu Lu, Jingyu Kuang, Tong Shao, Sisi Xie, Ming Li, Lingyun Zhu * and Lvyun Zhu *
1 Department of Biology and Chemistry, College of Sciences, National University of Defense Technology, Changsha 410073, China
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(17), 9862; https://doi.org/10.3390/ijms23179862 - 30 Aug 2022
Cited by 27 | Viewed by 6344
Abstract
Prime editing (PE), as a “search-and-replace” genome editing technology, has shown the attractive potential of versatile genome editing ability, which is, in principle, currently superior to other well-established genome-editing technologies in the all-in-one operation scope. However, essential technological solutions of PE technology, such [...] Read more.
Prime editing (PE), as a “search-and-replace” genome editing technology, has shown the attractive potential of versatile genome editing ability, which is, in principle, currently superior to other well-established genome-editing technologies in the all-in-one operation scope. However, essential technological solutions of PE technology, such as the improvement of genome editing efficiency, the inhibition of potential off-targets and intended edits accounting for unexpected side-effects, and the development of effective delivery systems, are necessary to broaden its application. Since the advent of PE, many optimizations have been performed on PE systems to improve their performance, resulting in bright prospects for application in many fields. This review briefly discusses the development of PE technology, including its functional principle, noteworthy barriers restraining its application, current efforts in technical optimization, and its application directions and potential risks. This review may provide a concise and informative insight into the burgeoning field of PE, highlight the exciting prospects for this powerful tool, and provide clues for questions that may propel the field forward. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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12 pages, 1333 KiB  
Article
Establishment and Characterization of Novel Human Intestinal In Vitro Models for Absorption and First-Pass Metabolism Studies
by Randy Przybylla 1, Christina Susanne Mullins 1, Mathias Krohn 1, Stefan Oswald 2 and Michael Linnebacher 1,*
1 Molecular Oncology and Immunotherapy, Clinic of General Surgery, 18057 Rostock, Germany
2 Institute of Pharmacology and Toxicology, Rostock University Medical Center, 18057 Rostock, Germany
Int. J. Mol. Sci. 2022, 23(17), 9861; https://doi.org/10.3390/ijms23179861 - 30 Aug 2022
Cited by 7 | Viewed by 2883
Abstract
Commonly used intestinal in vitro models are limited in their potential to predict oral drug absorption. They either lack the capability to form a tight cellular monolayer mimicking the intestinal epithelial barrier or the expression of cytochrome P450 3A4 (CYP3A4). The aim of [...] Read more.
Commonly used intestinal in vitro models are limited in their potential to predict oral drug absorption. They either lack the capability to form a tight cellular monolayer mimicking the intestinal epithelial barrier or the expression of cytochrome P450 3A4 (CYP3A4). The aim of this study was to establish a platform of colorectal cancer patient-derived cell lines for evaluation of human intestinal drug absorption and metabolism. We characterized ten 2D cell lines out of our collection with confluent outgrowth and long-lasting barrier forming potential as well as suitability for high throughput applications with special emphasis on expression and inducibility of CYP3A4. By assessment of the transepithelial electrical resistance (TEER) the cells barrier function capacity can be quantified. Very high TEER levels were detected for HROC60. A high basal CYP3A4 expression and function was found for HROC32. Eight cell lines showed higher CYP3A4 induction by stimulation via the vitamin D receptor compared to Caco-2 cells (5.1- to 16.8-fold change). Stimulation of the pregnane X receptor led to higher CYP3A4 induction in two cell lines. In sum, we identified the two cell lines HROC183 T0 M2 and HROC217 T1 M2 as useful tools for in vitro drug absorption studies. Due to their high TEER values and inducibility by drug receptor ligands, they may be superior to Caco-2 cells to analyze oral drug absorption and intestinal drug–drug interactions. Significance statement: Selecting appropriate candidates is important in preclinical drug development. Therefore, cell models to predict absorption from the human intestine are of the utmost importance. This study revealed that the human cell lines HROC183 T0 M2 and HROC217 T1 M2 may be better suited models and possess higher predictive power of pregnane X receptor- and vitamin D-mediated drug metabolism than Caco-2 cells. Consequently, they represent useful tools for predicting intestinal absorption and simultaneously enable assessment of membrane permeability and first-pass metabolism. Full article
(This article belongs to the Special Issue Biological Interfaces in Gastrointestinal Cancer 2.0)
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13 pages, 1393 KiB  
Article
Sensitivity of the Transport of Plastic Nanoparticles to Typical Phosphates Associated with Ionic Strength and Solution pH
by Xingyu Liu, Yan Liang *, Yongtao Peng, Tingting Meng, Liling Xu and Pengcheng Dong
School of Resources, Environment and Materials, Guangxi University, Nanning 530004, China
Int. J. Mol. Sci. 2022, 23(17), 9860; https://doi.org/10.3390/ijms23179860 - 30 Aug 2022
Cited by 3 | Viewed by 1939
Abstract
The influence of phosphates on the transport of plastic particles in porous media is environmentally relevant due to their ubiquitous coexistence in the subsurface environment. This study investigated the transport of plastic nanoparticles (PNPs) via column experiments, paired with Derjaguin–Landau–Verwey–Overbeek calculations and numerical [...] Read more.
The influence of phosphates on the transport of plastic particles in porous media is environmentally relevant due to their ubiquitous coexistence in the subsurface environment. This study investigated the transport of plastic nanoparticles (PNPs) via column experiments, paired with Derjaguin–Landau–Verwey–Overbeek calculations and numerical simulations. The trends of PNP transport vary with increasing concentrations of NaH2PO4 and Na2HPO4 due to the coupled effects of increased electrostatic repulsion, the competition for retention sites, and the compression of the double layer. Higher pH tends to increase PNP transport due to the enhanced deprotonation of surfaces. The release of retained PNPs under reduced IS and increased pH is limited because most of the PNPs were irreversibly captured in deep primary minima. The presence of physicochemical heterogeneities on solid surfaces can reduce PNP transport and increase the sensitivity of the transport to IS. Furthermore, variations in the hydrogen bonding when the two phosphates act as proton donors will result in different influences on PNP transport at the same IS. This study highlights the sensitivity of PNP transport to phosphates associated with the solution chemistries (e.g., IS and pH) and is helpful for better understanding the fate of PNPs and other colloidal contaminants in the subsurface environment. Full article
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10 pages, 2704 KiB  
Article
Improving the Laboratory Diagnosis of M-like Variants Related to Alpha1-Antitrypsin Deficiency
by Valentina Barzon 1, Stefania Ottaviani 2,*, Alice Maria Balderacchi 2, Alessandra Corino 2, Davide Piloni 2, Giulia Accordino 2, Manuela Coretti 2, Francesca Mariani 2, Angelo Guido Corsico 1,2 and Ilaria Ferrarotti 2
1 Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, University of Pavia, 27100 Pavia, Italy
2 Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, Laboratory of Biochemistry and Genetics, Pulmonology Unit, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy
Int. J. Mol. Sci. 2022, 23(17), 9859; https://doi.org/10.3390/ijms23179859 - 30 Aug 2022
Cited by 7 | Viewed by 2636
Abstract
Alpha1-antitrypsin (AAT) is a serine protease inhibitor that is encoded by the highly polymorphic SERPINA1 gene. Mutations in this gene can lead to AAT deficiency (AATD), which is associated with an increased risk of lung and/or liver disease. On the basis of electrophoretic [...] Read more.
Alpha1-antitrypsin (AAT) is a serine protease inhibitor that is encoded by the highly polymorphic SERPINA1 gene. Mutations in this gene can lead to AAT deficiency (AATD), which is associated with an increased risk of lung and/or liver disease. On the basis of electrophoretic migration, AAT variants are named with capital letters; M (medium) signifies the normal protein. Among pathological variants, the M-like ones represent a heterogeneous group of rare allelic variants that exhibit the same electrophoretic pattern as the M wild-type protein, which makes them difficult to detect with routine methods. In order to avoid their misdiagnosis, the present study defines and validates effective methods for the detection of two pathogenic M-like variants, Mwurzburg and Mwhitstable. Comparison of protein phenotypes using isoelectric focusing of samples that presented the Mwurzburg variant, as revealed by exons 5 sequencing, identified a particular electrophoretic pattern amenable to the Mwurzburg protein. The specific phenotyping pattern was retrospectively validated, thus enabling the detection of 16 patients with Mwurzburg variant among the subjects already tested but not sequenced according to our diagnostic algorithm. The Mwhitstable allele was detected by intron 4 sequencing of SERPINA1 gene. Mwurzburg and Mwhitstable are often misdiagnosed and the introduction of diagnostic improvements can help the clinical management, especially in patients with established lung disease without any other reported risk factors. Full article
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11 pages, 678 KiB  
Article
MMP-9 Levels in the Gingival Crevicular Fluid of Chilean Rosacea Patients
by Javier Fernández 1, Constanza Jiménez 2, Dafna Benadof 2, Paulina Morales 2, Jessica Astorga 3, Felipe Cáceres 2, Marcela Hernández 3,4, Alejandra Fernández 2,* and Fernando Valenzuela 5,*
1 Centro Internacional de Estudios Clínicos, Probity Medical Research, Santiago 8420383, Chile
2 Department of Oral Pathology, Faculty of Dentistry, Universidad Andres Bello, Santiago 8370133, Chile
3 Laboratory of Periodontal Biology, Faculty of Dentistry, Universidad de Chile, Santiago 8380544, Chile
4 Department of Oral Pathology and Medicine, Faculty of Dentistry, Universidad de Chile, Santiago 8380544, Chile
5 Department of Dermatology, Faculty of Medicine, Universidad de Los Andes, Av. Plaza 2501, Las Condes, Santiago 7620157, Chile
Int. J. Mol. Sci. 2022, 23(17), 9858; https://doi.org/10.3390/ijms23179858 - 30 Aug 2022
Cited by 9 | Viewed by 2394
Abstract
Rosacea is a chronic inflammatory skin disease whose prevalence rates remain unknown in Chile. Laboratory benchmark testing for this disease is not useful, therefore, we aimed to evaluate the gingival crevicular fluid (GCF) levels of extracellular metalloproteinases (MMP)-2 and MMP-9 as novel rosacea [...] Read more.
Rosacea is a chronic inflammatory skin disease whose prevalence rates remain unknown in Chile. Laboratory benchmark testing for this disease is not useful, therefore, we aimed to evaluate the gingival crevicular fluid (GCF) levels of extracellular metalloproteinases (MMP)-2 and MMP-9 as novel rosacea biomarkers. We designed a cross-sectional study with a control group. Participants were systemically healthy adults (n = 20) and persons with rosacea (n = 18). We performed a periodontal evaluation and collected gingival crevicular fluid to measure MMP-2 and MMP-9 levels. Analysis showed mean and standard deviation of MMP-9 concentrations in the GCF for patients with rosacea was 764.52 ± 569.83 pg/mL; for healthy patients, it was 260.69 ± 170.43 pg/mL (p < 0.05). The diagnosis of rosacea was responsible for the levels of MMP-9 in the GCF (p < 0.05), as opposed to periodontitis, smoking, and age (p > 0.05). The Area under ROC for MMP-9 was 0.869 (95%, C.I: 0.719–0.956), with a sensitivity of 72.22% and specificity of 81.58% for the diagnosis of rosacea. We conclude that the quantification of MMP-9 in the GCF could be used as a biomarker of rosacea. Also, rosacea was responsible for increasing the levels of MMP-9 in the GCF independent of periodontal status. Full article
(This article belongs to the Special Issue State-of-the-Art Molecular Immunology in Chile)
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20 pages, 3366 KiB  
Article
Staphylococcus aureus Exfoliative Toxin E, Oligomeric State and Flip of P186: Implications for Its Action Mechanism
by Carolina Gismene 1,†, Jorge Enrique Hernández González 1,†, Angela Rocio Niño Santisteban 1, Andrey Fabricio Ziem Nascimento 2, Lucas dos Santos Cunha 3, Fábio Rogério de Moraes 1, Cristiano Luis Pinto de Oliveira 4, Caio C. Oliveira 3, Paola Jocelan Scarin Provazzi 5, Pedro Geraldo Pascutti 6, Raghuvir Krishnaswamy Arni 1,* and Ricardo Barros Mariutti 1,*
1 Multiuser Center for Biomolecular Innovation, IBILCE/UNESP, São José do Rio Preto 15054-000, Brazil
2 Brazilian Synchrotron Light Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas 13083-970, Brazil
3 Institute of Chemistry, University of Campinas, Campinas 13083-970, Brazil
4 Instituto de Física, University of São Paulo, São Paulo 05314-970, Brazil
5 Laboratory of Genomic Studies, Sao Paulo State University-UNESP, São José do Rio Preto 15054-000, Brazil
6 Laboratório de Modelagem e Dinâmica Molecular, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-901, Brazil
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(17), 9857; https://doi.org/10.3390/ijms23179857 - 30 Aug 2022
Cited by 5 | Viewed by 3290
Abstract
Staphylococcal exfoliative toxins (ETs) are glutamyl endopeptidases that specifically cleave the Glu381-Gly382 bond in the ectodomains of desmoglein 1 (Dsg1) via complex action mechanisms. To date, four ETs have been identified in different Staphylococcus aureus strains and ETE is the most recently characterized. [...] Read more.
Staphylococcal exfoliative toxins (ETs) are glutamyl endopeptidases that specifically cleave the Glu381-Gly382 bond in the ectodomains of desmoglein 1 (Dsg1) via complex action mechanisms. To date, four ETs have been identified in different Staphylococcus aureus strains and ETE is the most recently characterized. The unusual properties of ETs have been attributed to a unique structural feature, i.e., the 180° flip of the carbonyl oxygen (O) of the nonconserved residue 192/186 (ETA/ETE numbering), not conducive to the oxyanion hole formation. We report the crystal structure of ETE determined at 1.61 Å resolution, in which P186(O) adopts two conformations displaying a 180° rotation. This finding, together with free energy calculations, supports the existence of a dynamic transition between the conformations under the tested conditions. Moreover, enzymatic assays showed no significant differences in the esterolytic efficiency of ETE and ETE/P186G, a mutant predicted to possess a functional oxyanion hole, thus downplaying the influence of the flip on the activity. Finally, we observed the formation of ETE homodimers in solution and the predicted homodimeric structure revealed the participation of a characteristic nonconserved loop in the interface and the partial occlusion of the protein active site, suggesting that monomerization is required for enzymatic activity. Full article
(This article belongs to the Special Issue Protein Structure Research)
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