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Int. J. Mol. Sci., Volume 9, Issue 11 (November 2008), Pages 2062-2321

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Research

Jump to: Review

Open AccessArticle Terminal Continuation (TC) RNA Amplification Enables Expression Profiling Using Minute RNA Input Obtained from Mouse Brain
Int. J. Mol. Sci. 2008, 9(11), 2091-2104; doi:10.3390/ijms9112091
Received: 16 September 2008 / Revised: 17 October 2008 / Accepted: 23 October 2008 / Published: 31 October 2008
Cited by 18 | PDF Full-text (2384 KB) | HTML Full-text | XML Full-text
Abstract
A novel methodology named terminal continuation (TC) RNA amplification has been developed to amplify RNA from minute amounts of starting material. Utility of the TC RNA amplification method is demonstrated with two new modifications including obviating the need for second strand synthesis, [...] Read more.
A novel methodology named terminal continuation (TC) RNA amplification has been developed to amplify RNA from minute amounts of starting material. Utility of the TC RNA amplification method is demonstrated with two new modifications including obviating the need for second strand synthesis, and purifying the amplification template using column filtration prior to in vitro transcription (IVT). Using four low concentrations of RNA extracted from mouse brain (1, 10, 25 and 50 ng), one round TC RNA amplification was compared to one round amplified antisense RNA (aRNA) in conjunction with column filtration and drop dialysis purification. The TC RNA amplification without second strand synthesis performed extremely well on customdesigned cDNA array platforms, and column filtration was found to provide higher positive detection of individual clones when hybridization signal intensity was subtracted from corresponding negative control hybridization signal levels. Results indicate that TC RNA amplification without second strand synthesis, in conjunction with column filtration, is an excellent method for RNA amplification from extremely small amounts of input RNA from mouse brain and postmortem human brain, and is compatible with microaspiration strategies and subsequent microarray analysis. Full article
(This article belongs to the Special Issue Advances in Molecular Neuropathology)
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Open AccessArticle TFIP11 Interacts with mDEAH9, an RNA Helicase Involved in Spliceosome Disassembly
Int. J. Mol. Sci. 2008, 9(11), 2105-2113; doi:10.3390/ijms9112105
Received: 18 August 2008 / Revised: 31 October 2008 / Accepted: 3 November 2008 / Published: 4 November 2008
Cited by 10 | PDF Full-text (516 KB) | HTML Full-text | XML Full-text
Abstract
Yeast proteins Ntr1, Ntr2 and Prp43 function in spliceosome disassembly. An Ntr1-Ntr2 protein complex recruits Prp43 to allow the removal of the lariat-intron in latestage RNA splicing activity. Based on amino-acid sequence similarities across species, TFIP11 and mDEAH9/Dhx15 have been identified as [...] Read more.
Yeast proteins Ntr1, Ntr2 and Prp43 function in spliceosome disassembly. An Ntr1-Ntr2 protein complex recruits Prp43 to allow the removal of the lariat-intron in latestage RNA splicing activity. Based on amino-acid sequence similarities across species, TFIP11 and mDEAH9/Dhx15 have been identified as homologues of yeast Ntr1 and Prp43, respectively. The N-terminal region of TFIP11 contains a G-patch, which is a highly conserved domain of many RNA-processing proteins. TFIP11 displays a unique and characteristic subnuclear localization pattern, in close proximity to SC35 nuclear speckles. Transfected GFP-tagged mDEAH9 displays an evenly distributed nuclear localization and is excluded from the nucleoli; however when TFIP11 and mDEAH9 are co-transfected, both proteins colocalize to distinct nuclear speckles. These data show that TFIP11 recruits mDEAH9 suggesting that these two proteins have similar biological activities to their yeast counterparts. Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Open AccessArticle Occurrence of Deoxynivalenol in Maize and Wheat in Serbia
Int. J. Mol. Sci. 2008, 9(11), 2114-2126; doi:10.3390/ijms9112114
Received: 21 July 2008 / Revised: 25 September 2008 / Accepted: 27 September 2008 / Published: 4 November 2008
Cited by 19 | PDF Full-text (256 KB) | HTML Full-text | XML Full-text
Abstract
A total of 226 samples of maize and 59 of wheat from the 2004−2007 harvests were investigated for the presence and concentration of deoxynivalenol (DON). Samples of the 2004 harvest were analyzed after their storing for one year in barns, while those [...] Read more.
A total of 226 samples of maize and 59 of wheat from the 2004−2007 harvests were investigated for the presence and concentration of deoxynivalenol (DON). Samples of the 2004 harvest were analyzed after their storing for one year in barns, while those of the 2005−2007 harvest were taken directly off fields immediately after the harvest. The samples were analyzed by liquid chromatography on an ODS Hypersil column with DAD detector and ELISA methods. The average incidence rate of DON in maize from the 2004 harvest was 50% (concentration range 0.042−2.460 mg/kg, average value 0.536 mg/kg), while for those of the 2005−2007 harvest it was 32.4% (concentration range 0.027−2.210 mg/kg, average value 0.223 mg/kg). In the case of wheat incidence rate of DON for 2004 harvest was 50.0% (concentration range 0.630−1.840 mg/kg, average value 1.235 mg/kg), while for those of the 2005−2007 harvest it was 34.5% (concentration range 0.057−0.423 mg/kg, average value 0.190 mg/kg). Concentrations in two samples of maize and one of wheat (one sample of each cereal being of the 2004 harvest) were above the maximum level adopted by the European Commission. The results obtained were analyzed as a function of climatic conditions and compared with those of the neighboring countries where the relevant data existed. Full article
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Open AccessArticle Synthesis and Properties of Poly[p-(2,5-dihydroxy)- phenylenebenzobisoxazole] Fiber
Int. J. Mol. Sci. 2008, 9(11), 2159-2168; doi:10.3390/ijms9112159
Received: 13 July 2008 / Revised: 27 October 2008 / Accepted: 29 October 2008 / Published: 5 November 2008
Cited by 7 | PDF Full-text (404 KB) | HTML Full-text | XML Full-text
Abstract
The novel polymer poly[p-(2,5-dihydroxy)-phenylenebenzobisoxazole] (PBOH) fiber was synthesized in the presence of 2,5-dihydroxyterephthalicacid (DHTA) and 4,6-diamino-1,3-benzenediol in poly(phosphoric acid) (PPA) using typical polycondensation conditions. The crystalline solutions of liquid PBOH in PPA were spun into fibers using dry-jet wet spinning. [...] Read more.
The novel polymer poly[p-(2,5-dihydroxy)-phenylenebenzobisoxazole] (PBOH) fiber was synthesized in the presence of 2,5-dihydroxyterephthalicacid (DHTA) and 4,6-diamino-1,3-benzenediol in poly(phosphoric acid) (PPA) using typical polycondensation conditions. The crystalline solutions of liquid PBOH in PPA were spun into fibers using dry-jet wet spinning. Furthermore, the thermostability and mechanical properties of PBOH were compared with poly(p-phenylene-2,6-benzoxazole) (PBO) in order to investigate the relationship between the chain structure and properties. The results indicated that the thermal degradation temperature of PBOH was above 750K and the tensile strength of the PBOH fiber was 3.1GPa, which were much lower than those of PBO fiber. The compressive strength of PBOH fiber was 331 M Pa, which was slightly higher than that of PBO fiber. In addition, molecular simulation was employed to explain why the compressive strength of PBOH fiber did not increase significantly compared to PBO fiber. Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Open AccessArticle Survey of Slaughtered Pigs for Occurrence of Ochratoxin A and Porcine Nephropathy in Serbia
Int. J. Mol. Sci. 2008, 9(11), 2169-2183; doi:10.3390/ijms9112169
Received: 19 March 2008 / Revised: 29 October 2008 / Accepted: 31 October 2008 / Published: 7 November 2008
Cited by 25 | PDF Full-text (292 KB) | HTML Full-text | XML Full-text
Abstract
Samples of blood, kidney and liver were randomly selected from slaughtered pigs (n=90) and analyzed for ochratoxin A by HPLC. In addition, in order to obtain information on the occurrence of nephropathy, histological examinations were carried out. Of the 90 liver samples, [...] Read more.
Samples of blood, kidney and liver were randomly selected from slaughtered pigs (n=90) and analyzed for ochratoxin A by HPLC. In addition, in order to obtain information on the occurrence of nephropathy, histological examinations were carried out. Of the 90 liver samples, 26.6% contained OTA in the range of 0.22-14.5 ng/g. The incidence of OTA in serum and kidney were very similar (31%, 33.3%), with a maximum concentration of 220.8 ng/mL, and 52.5 ng/g, respectively. Histopathological examination of kidneys confirmed tubulopathies with edema and cell vacuolization. In addition, hemorrhages and necrosis of proximal kidney tubules’ cells were found. Full article
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Open AccessCommunication Effect of Ammonia on the Gas-Phase Hydration of the Common Atmospheric Ion HSO4-
Int. J. Mol. Sci. 2008, 9(11), 2184-2193; doi:10.3390/ijms9112184
Received: 23 May 2008 / Revised: 20 October 2008 / Accepted: 23 October 2008 / Published: 7 November 2008
Cited by 16 | PDF Full-text (408 KB) | HTML Full-text | XML Full-text
Abstract
Hydration directly affects the mobility, thermodynamic properties, lifetime and nucleation rates of atmospheric ions. In the present study, the role of ammonia on the formation of hydrogen bonded complexes of the common atmospheric hydrogensulfate (HSO4-) ion with water has [...] Read more.
Hydration directly affects the mobility, thermodynamic properties, lifetime and nucleation rates of atmospheric ions. In the present study, the role of ammonia on the formation of hydrogen bonded complexes of the common atmospheric hydrogensulfate (HSO4-) ion with water has been investigated using the Density Functional Theory (DFT). Our findings rule out the stabilizing effect of ammonia on the formation of negatively charged cluster hydrates and show clearly that the conventional (classical) treatment of ionic clusters as presumably more stable compared to neutrals may not be applicable to pre-nucleation clusters. These considerations lead us to conclude that not only quantitative but also qualitative assessment of the relative thermodynamic stability of atmospheric clusters requires a quantum-chemical treatment. Full article
(This article belongs to the Special Issue The Chemical Bond and Bonding)
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Open AccessArticle Detection of FLT3 Oncogene Mutations in Acute Myeloid Leukemia Using Conformation Sensitive Gel Electrophoresis
Int. J. Mol. Sci. 2008, 9(11), 2194-2204; doi:10.3390/ijms9112194
Received: 13 June 2008 / Revised: 27 October 2008 / Accepted: 4 November 2008 / Published: 11 November 2008
Cited by 7 | PDF Full-text (232 KB) | HTML Full-text | XML Full-text
Abstract
FLT3 (fms-related tyrosine kinase 3) is a receptor tyrosine kinase class III that is expressed on by early hematopoietic progenitor cells and plays an important role in hematopoietic stem cell proliferation, differentiation and survival. FLT3 is also expressed on leukemia blasts in [...] Read more.
FLT3 (fms-related tyrosine kinase 3) is a receptor tyrosine kinase class III that is expressed on by early hematopoietic progenitor cells and plays an important role in hematopoietic stem cell proliferation, differentiation and survival. FLT3 is also expressed on leukemia blasts in most cases of acute myeloid leukemia (AML). In order to determine the frequency of FLT3 oncogene mutations, we analyzed genomic DNA of adult de novo acute myeloid leukemia (AML). Polymerase chain reaction (PCR) and conformation-sensitive gel electrophoresis (CSGE) were used for FLT3 exons 11, 14, and 15, followed by direct DNA sequencing. Two different types of functionally important FLT 3 mutations have been identified. Those mutations were unique to patients with inv(16), t(15:17) or t(8;21) and comprised fifteen cases with internal tandem duplication (ITD) mutation in the juxtamembrane domain and eleven cases with point mutation (exon 20, Asp835Tyr). The high frequency of the flt3 proto-oncogene mutations in acute myeloid leukemia AML suggests a key role for the receptor function. The association of FLT3 mutations with chromosomal abnormalities invites speculation as to the link between these two changes in the pathogenesis of acute myeloid leukemiaAML. Furthermore, CSGE method has shown to be a rapid and sensitive screening method for detection of nucleotide alteration in FLT3 gene. Finally, this study reports, for the first time in Saudi Arabia, mutations in the human FLT3 gene in acute myeloid leukemia AML patients. Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Open AccessArticle Intestinal Structure and Function of Broiler Chickens on Diets Supplemented with a Synbiotic Containing Enterococcus faecium and Oligosaccharides
Int. J. Mol. Sci. 2008, 9(11), 2205-2216; doi:10.3390/ijms9112205
Received: 16 September 2008 / Revised: 31 October 2008 / Accepted: 7 November 2008 / Published: 12 November 2008
Cited by 29 | PDF Full-text (124 KB) | HTML Full-text | XML Full-text
Abstract
A feeding trial was conducted on broiler chickens to study the effects of the synbiotic BIOMIN IMBO [a combination of Enterococcus faecium, a prebiotic (derived from chicory) and immune modulating substances (derived from sea algae)], with a dose of 1 kg/ton of the starter diets and 0.5 kg/ton of the grower diets on the intestinal morphometry and nutrient absorption. The general performance was improved (P < 0.05) by the dietary inclusion of synbiotic compared with the controls. Furthermore, the addition of synbiotic increased (P < 0.001) the villus height/crypt depth ratio and villus height in ileum. However, the ileal crypt depth was decreased by dietary supplementation of synbiotic compared with control. The addition of glucose in Ussing chamber produced a significant increase (P ≤ 0.001) in short-circuit current (Isc) in jejunum and colon relative to the basal values in both synbiotic and control groups. However, in jejunum the percentage of Isc increase after glucose addition was higher for synbiotic group (333 %) than control group (45 %). In conclusion, dietary inclusion of synbiotic BIOMIN IMBO increased the growth performance and improved intestinal morphology and nutrient absorption. Full article
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Open AccessArticle Extended Grunwald-Winstein Analysis - LFER Used to Gauge Solvent Effects in p-Nitrophenyl Chloroformate Solvolysis
Int. J. Mol. Sci. 2008, 9(11), 2231-2242; doi:10.3390/ijms9112231
Received: 27 October 2008 / Revised: 9 November 2008 / Accepted: 12 November 2008 / Published: 13 November 2008
Cited by 14 | PDF Full-text (233 KB) | HTML Full-text | XML Full-text
Abstract
Specific rates of solvolysis at 25oC for p-nitrophenyl chloroformate (1) are analyzed using the extended (two-term) Grunwald-Winstein equation. For 39 solvents, the sensitivities (l = 1.68±0.06 and m = 0.46±0.04) towards changes in solvent nucleophilicity (l) [...] Read more.
Specific rates of solvolysis at 25oC for p-nitrophenyl chloroformate (1) are analyzed using the extended (two-term) Grunwald-Winstein equation. For 39 solvents, the sensitivities (l = 1.68±0.06 and m = 0.46±0.04) towards changes in solvent nucleophilicity (l) and solvent ionizing power (m) obtained, are similar to those previously observed for phenyl chloroformate (2) and p-methoxyphenyl chloroformate (3). The observations incorporating new kinetic data in several fluoroalcohol-containing mixtures, are rationalized in terms of the reaction being sensitive to substituent effects and the mechanism of reaction involving the addition (association) step of an additionelimination (association-dissociation) pathway being rate-determining. The l/m ratios obtained for 1, 2, and 3, are also compared to the previously published l/m ratios for benzyl chloroformate (4) and p-nitrobenzyl chloroformate (5). Full article
(This article belongs to the Special Issue Grunwald-Winstein Equations – 60 Years & Counting)
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Open AccessArticle Effect of Several New and Currently Available Oxime Cholinesterase Reactivators on Tabun-intoxicated Rats
Int. J. Mol. Sci. 2008, 9(11), 2243-2252; doi:10.3390/ijms9112243
Received: 16 October 2008 / Revised: 4 November 2008 / Accepted: 7 November 2008 / Published: 14 November 2008
Cited by 11 | PDF Full-text (65 KB) | HTML Full-text | XML Full-text
Abstract
The therapeutical efficacies of eleven oxime-based acetylcholinesterase reactivators were compared in an in vivo (rat model) study of treatment of intoxication caused by tabun. In this group there were some currently available oximes (obidoxime, trimedoxime and HI-6) and the rest were newly [...] Read more.
The therapeutical efficacies of eleven oxime-based acetylcholinesterase reactivators were compared in an in vivo (rat model) study of treatment of intoxication caused by tabun. In this group there were some currently available oximes (obidoxime, trimedoxime and HI-6) and the rest were newly synthesized compounds. The best reactivation efficacy for acetylcholinesterase in blood (expressed as percent of reactivation) among the currently available oximes was observed after administration of trimedoxime (16%) and of the newly synthesized K127 (22432) (25%). The reactivation of butyrylcholinesterase in plasma was also studied; the best reactivators were trimedoxime, K117 (22435), and K127 (22432), with overall reactivation efficacies of approximately 30%. Partial protection of brain ChE against tabun inhibition was observed after administration of trimedoxime (acetylcholinesterase 20%; butyrylcholinesterase 30%) and obidoxime (acetylcholinesterase 12%; butyrylcholinesterase 16%). Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
Open AccessArticle Pravastatin Prevents Aortic Atherosclerosis via Modulation of Signal Transduction and Activation of Transcription 3 (STAT3) to Attenuate Interleukin-6 (IL-6) Action in ApoE Knockout Mice
Int. J. Mol. Sci. 2008, 9(11), 2253-2264; doi:10.3390/ijms9112253
Received: 27 September 2008 / Revised: 4 November 2008 / Accepted: 7 November 2008 / Published: 14 November 2008
Cited by 9 | PDF Full-text (269 KB) | HTML Full-text | XML Full-text
Abstract
The purpose of this study was to determine whether pravastatin’s prevention of aortic atherosclerosis via attenuation of IL-6 action depends on modulation of STAT3 activity. Male apoE knockout (apoE-/-) mice fed on a diet containing 1.25% cholesterol (wt/wt) were divided into pravastatin [...] Read more.
The purpose of this study was to determine whether pravastatin’s prevention of aortic atherosclerosis via attenuation of IL-6 action depends on modulation of STAT3 activity. Male apoE knockout (apoE-/-) mice fed on a diet containing 1.25% cholesterol (wt/wt) were divided into pravastatin group provided with pravastatin (80 mg kg-1 per day) and atherosclerosis group. After eight weeks, pravastatin significantly prevented atherosclerotic lesion and reduced levels of IL-6 in serum and lesion, and significantly decreased expressions of phosphorylated STAT3 (pSTAT3) and increased suppressor of cytokine signaling 3 (SOCS3) expressions in lesions. Our results suggested that pravastatin’s aortic atherosclerosis preventing action via attenuation of IL-6 action may partially depend on modulation of STAT3 activity. Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
Open AccessArticle Ponicidin Inhibits Monocytic Leukemia Cell Growth by Induction of Apoptosis
Int. J. Mol. Sci. 2008, 9(11), 2265-2277; doi:10.3390/ijms9112265
Received: 23 September 2008 / Revised: 25 October 2008 / Accepted: 7 November 2008 / Published: 19 November 2008
Cited by 8 | PDF Full-text (353 KB) | HTML Full-text | XML Full-text
Abstract
In this study two monocytic leukemia cell lines, U937 and THP-1 cells, were used to investigate the anti-proliferation effects caused by ponicidin. Cell viability was measured by an MTT assay. Cell apoptosis was assessed by flow cytometry as well as DNA fragmentation [...] Read more.
In this study two monocytic leukemia cell lines, U937 and THP-1 cells, were used to investigate the anti-proliferation effects caused by ponicidin. Cell viability was measured by an MTT assay. Cell apoptosis was assessed by flow cytometry as well as DNA fragmentation analysis. Cell morphology was observed using an inverted microscope and Hoechst 33258 staining. RT-PCR and Western blot analysis were used to detect survivin as well as Bax and Bcl-2 expressions after the cells were treated with different concentrations of ponicidin. The results revealed that ponicidin could inhibit the growth of U937 and THP-1 cells significantly by induction of apoptosis. The suppression was in both time- and dose-dependent manner. Marked morphological changes of cell apoptosis were observed clearly after the cells were treated with ponicidin for 48~72 h. RT-PCR and Western blot analysis demonstrated that both survivin and Bcl-2 expressions were down-regulated remarkably while Bax expression remained constant before and after apoptosis occurred. We therefore conclude that ponicidin has significant anti-proliferation effects by inducing apoptosis on leukemia cells in vitro, downregulation of survivin as well as Bcl-2 expressions may be the important apoptosis inducing mechanisms. The results suggest that ponicidin may serve as potential therapeutic agent for leukemia. Full article
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
Open AccessArticle A Novel Co-polymer Based on Hydroxypropyl α-Cyclodextrin Conjugated to Low Molecular Weight Polyethylenimine as an in Vitro Gene Delivery Vector
Int. J. Mol. Sci. 2008, 9(11), 2278-2289; doi:10.3390/ijms9112278
Received: 31 May 2008 / Revised: 4 November 2008 / Accepted: 7 November 2008 / Published: 21 November 2008
Cited by 12 | PDF Full-text (578 KB) | HTML Full-text | XML Full-text
Abstract
A novel co-polymer based on 2-hydroxypropyl-α-cyclodextrin cross-linked by low molecular weight polyethylenimine was synthesized as a gene delivery vector. The copolymer could bind and condense DNA tightly. It showed lower cytotoxicity than PEI 25kDa in SK-BR-3 cells. Transfection efficiency was increased over [...] Read more.
A novel co-polymer based on 2-hydroxypropyl-α-cyclodextrin cross-linked by low molecular weight polyethylenimine was synthesized as a gene delivery vector. The copolymer could bind and condense DNA tightly. It showed lower cytotoxicity than PEI 25kDa in SK-BR-3 cells. Transfection efficiency was increased over 5.5-fold higher than PEI 25 kDa in SK-BR-3 cells in complete serum medium. It is a potential candidate vector for gene therapy. Full article
Open AccessArticle Simulation of Doxorubicin Delivery via Glucosamine(ethylene glycol) Carrier
Int. J. Mol. Sci. 2008, 9(11), 2290-2305; doi:10.3390/ijms9112290
Received: 20 August 2008 / Revised: 2 October 2008 / Accepted: 13 November 2008 / Published: 21 November 2008
PDF Full-text (552 KB) | HTML Full-text | XML Full-text
Abstract
This article focuses on the molecular modeling of the release of doxorubicin from capsules composed of glucosamine(ethylene glycol) oligomers. Doxorubicin forms micelle structures with glucosamine(ethylene glycol), and the drug release mechanism can be studied through the modeling of oligomeric bond breaking under [...] Read more.
This article focuses on the molecular modeling of the release of doxorubicin from capsules composed of glucosamine(ethylene glycol) oligomers. Doxorubicin forms micelle structures with glucosamine(ethylene glycol), and the drug release mechanism can be studied through the modeling of oligomeric bond breaking under acidic, neutral, or basic conditions. Under these conditions, the activation energies were calculated to be 145.51, 135.78, and 287.60 kcal/mol, respectively, at the B3LYP/6-31G//PM3 level. Based on these values, doxorubicin can be released into acidic and neutral solutions but not into basic solution. Ethylene glycol chain length in glucosamine(ethylene glycol) also effects drug release. As the length of ethylene glycol increases, the amount of drug released increases under acidic conditions, but decreases under neutral and basic conditions. When the drug is released from glucosamine(ethylene glycol) oligomers, the drug molecule and glucosamine(ethylene glycol) molecules form a micelle structure. Studies found that, as the length of the ethylene glycol chains increases, the micelle structure is more easily formed. The ethylene glycol group can deliver doxorubicin to cancer cells in micelle form. Full article
(This article belongs to the Section Physical Chemistry, Theoretical and Computational Chemistry)
Open AccessArticle Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error
Int. J. Mol. Sci. 2008, 9(11), 2306-2321; doi:10.3390/ijms9112306
Received: 28 August 2008 / Revised: 19 November 2008 / Accepted: 24 November 2008 / Published: 25 November 2008
Cited by 39 | PDF Full-text (362 KB) | HTML Full-text | XML Full-text
Abstract
Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W.G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in [...] Read more.
Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W.G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in plant tissue. We demonstrated that increasing the amount of plant material used for DNA extraction to 0.5-1.0 g considerably reduced sampling error and improved the reproducibility of DNA yield. The costs of DNA extraction at different scales and with different methods (commercial kits versus cetyltrimethylammonium bromide-based protocol) and qPCR systems (doubly labeled hybridization probes versus SYBR Green) were compared. A cost-effective protocol for the quantification of F. graminearum and F. culmorum DNA in wheat grain and maize stalk debris based on DNA extraction from 0.5-1.0 g material and real-time PCR with SYBR Green fluorescence detection was developed. Full article

Review

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Open AccessReview Fumonisins, Trichothecenes and Zearalenone in Cereals
Int. J. Mol. Sci. 2008, 9(11), 2062-2090; doi:10.3390/ijms9112062
Received: 28 July 2008 / Revised: 26 October 2008 / Accepted: 28 October 2008 / Published: 31 October 2008
Cited by 89 | PDF Full-text (200 KB) | HTML Full-text | XML Full-text
Abstract
Fumonisins are phytotoxic mycotoxins which are synthesized by various species of the fungal genus Fusarium such as Fusarium verticillioides (Sacc.) Nirenberg (ex F.moniliforme Sheldon) and Fusarium proliferatum. The trichothecene (TC) mycotoxins are secondary metabolites produce by species that belong to several [...] Read more.
Fumonisins are phytotoxic mycotoxins which are synthesized by various species of the fungal genus Fusarium such as Fusarium verticillioides (Sacc.) Nirenberg (ex F.moniliforme Sheldon) and Fusarium proliferatum. The trichothecene (TC) mycotoxins are secondary metabolites produce by species that belong to several fungal genera, especially Fusarium, Stachybotrys, Trichothecium, Trichoderma, Memnoniella and Myrothecium. Fusarium mycotoxins are widely dispersed in cereals and their products. Zearalenone (ZEA) is an estrogenic compound produced by Fusarium spp. such as F. graminearum and F. culmorum. Fumonisins, the TCs and ZEA are hazardous for human and animal health. Contamination with TCs causes a number of illnesses in human and animal such as decrease in food consumption (anorexia), depression or inhibition on immune system function and haematoxicity. The purpose of this paper is to give a review of the papers published on the field of fumonisin, TC and ZEA mycotoxins in cereals consumed in the world. Full article
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Open AccessReview Effects of Milling and Cooking Processes on the Deoxynivalenol Content in Wheat
Int. J. Mol. Sci. 2008, 9(11), 2127-2145; doi:10.3390/ijms9112127
Received: 31 July 2008 / Revised: 29 October 2008 / Accepted: 3 November 2008 / Published: 5 November 2008
Cited by 58 | PDF Full-text (139 KB) | HTML Full-text | XML Full-text
Abstract
Deoxynivalenol (DON, vomitoxin) is a natural-occuring mycotoxin mainly produced by Fusarium graminearum, a food-borne fungi widely distributed in crops and it is one of the most important mycotoxins in wheat and wheat-based foods and feeds. DON affects animal and human health [...] Read more.
Deoxynivalenol (DON, vomitoxin) is a natural-occuring mycotoxin mainly produced by Fusarium graminearum, a food-borne fungi widely distributed in crops and it is one of the most important mycotoxins in wheat and wheat-based foods and feeds. DON affects animal and human health causing diarrhea, vomiting, gastro-intestinal inflammation, and immunomodulation. Since the rate of the occurrence of DON in wheat is high, effective procedures to remove or eliminate DON from food products is essential to minimize exposures in those who consume large amounts of wheat. Cleaning prior to milling reduced to some extent the concentration of DON in final products. Since DON is distributed throughout the kernels, with higher content in the outer skin, milling is also effective in reducing the DON levels of wheat-based foods if bran and shorts are removed before thermal cooking. DON is water-soluble and cooking with larger amounts of water lowers DON content in products such as spaghetti and noodles. During baking or heating, DON is partially degraded to DON-related chemicals, whose toxicological effects are not studied well. This paper reviews the researches on the effects of milling and cooking on the DON level and discusses the perspectives of further studies. Full article
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Open AccessReview T-2 Toxin-induced Toxicity in Pregnant Mice and Rats
Int. J. Mol. Sci. 2008, 9(11), 2146-2158; doi:10.3390/ijms9112146
Received: 17 October 2008 / Revised: 31 October 2008 / Accepted: 4 November 2008 / Published: 5 November 2008
Cited by 33 | PDF Full-text (2497 KB) | HTML Full-text | XML Full-text
Abstract
T-2 toxin is a cytotoxic secondary fungal metabolite that belongs to the trichothecene mycotoxin family. This mycotoxin is a well known inhibitor of protein synthesis through its high binding affinity to peptidyl transferase, which is an integral part of the ribosomal 60s [...] Read more.
T-2 toxin is a cytotoxic secondary fungal metabolite that belongs to the trichothecene mycotoxin family. This mycotoxin is a well known inhibitor of protein synthesis through its high binding affinity to peptidyl transferase, which is an integral part of the ribosomal 60s subunit, and it also inhibits the synthesis of DNA and RNA, probably secondary to the inhibition of protein synthesis. In addition, T-2 toxin is said to induce apoptosis in many types of cells bearing high proliferating activity. T-2 toxin readily passes the placenta and is distributed to embryo/fetal tissues, which include many component cells bearing high proliferating activity. This paper reviews the reported data related to T-2 toxin-induced maternal and fetal toxicities in pregnant mice and rats. The mechanisms of T-2 toxin-induced apoptosis in maternal and fetal tissues are also discussed in this paper. Full article
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Open AccessReview Contribution of Natural Inhibitors to the Understanding of the PI3K/PDK1/PKB Pathway in the Insulin-mediated Intracellular Signaling Cascade
Int. J. Mol. Sci. 2008, 9(11), 2217-2230; doi:10.3390/ijms9112217
Received: 15 July 2008 / Revised: 8 November 2008 / Accepted: 12 November 2008 / Published: 12 November 2008
Cited by 18 | PDF Full-text (387 KB) | HTML Full-text | XML Full-text
Abstract
The critical initial steps in insulin action include phosphorylation of adapter proteins and activation of phosphatidylinositol 3-kinase (PI3K). One of important components in this process is a protein called Akt/protein kinase B (PKB). The work of numerous different researchers indicates a role [...] Read more.
The critical initial steps in insulin action include phosphorylation of adapter proteins and activation of phosphatidylinositol 3-kinase (PI3K). One of important components in this process is a protein called Akt/protein kinase B (PKB). The work of numerous different researchers indicates a role of PKB in regulating insulin-stimulated glucose uptake. The crucial role of lipid second messengers in PKB activation has been dissected through the use of the PI3K-specific inhibitors wortmannin and LY294002. Receptor-activated PI3K synthesizes the lipid second messenger PtdIns[3,4,5]-trisphosphate, leading to the recruitment of PKB to the membrane. Membrane attachment of PKB is mediated by its pleckstrin homology domain binding to PtdIns[3,4,5]-trisphosphate or PtdIns[3,4]-bisphosphate with high affinity. Activation of PKB alpha is then achieved at the plasma membrane by phosphorylation of Thr308 in the activation-loop of the kinase domain and Ser473 in the carboxy-terminal regulatory region, respectively. 3-Phosphoinositide-dependent protein kinase-1 (PDK1) is responsible for T308 phosphorylation. The usage of specific inhibitors and natural compound has significantly contributed to investigate the molecular mechanism of PI3K/PDK1/PKB signaling pathway, leading to the putative therapeutics benefits of patients. This review focuses on the contribution of natural inhibitor or compound in our understanding of the mechanism by which insulin induces, especially in PI3K/ PDK1/PKB signaling. Full article
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