Correlation between the Chemical and Genetic Relationships among Thymus saturejoides Genotypes Cultured under in vitro and in vivo Environments

In this study, the in vitro and in vivo essential oil (EO) composition and genetic variability in six micropropagated genotypes of Thymus saturejoides Coss ., a Mediterranean medicinal and aromatic plant, were analyzed by GC/MS and randomly amplified polymorphic DNA (RAPD). Yield and composition of the EO varied between genotypes. Cluster analysis based on RAPD data and EO grouped the six genotypes in three groups in both culture conditions, thus showing considerable intraspecific genetic and chemical variations. Applying the Mantel test, the result showed a significant correlation between the two proximity matrices RAPD and EO obtained from in vitro genotypes, whereas this correlation was not observed when using the EO obtained from the in vivo genotypes.     

Meta-analysis to refine map position and reduce confidence intervals for delayed-canopy-wilting QTLs in soybean

Slow canopy wilting in soybean has been identified as a potentially beneficial trait for ameliorating drought effects on yield. Previous research identified QTLs for slow wilting from two different biparental populations, and this information was combined with data from three other populations to identify nine QTL clusters for slow wilting on Gm02, Gm05, Gm11, Gm 14, Gm17, and Gm19. The QTL cluster on Gm14 was eliminated because these QTLs appeared to be false positives. In the present research, QTLs from these remaining eight clusters were compiled onto the soybean consensus map for meta-QTL analysis. Five model selection criteria were used to determine the most appropriate number of meta-QTLs at these eight chromosomal regions. For a QTL cluster on Gm02, two meta-QTLs were identified, whereas for the remaining seven QTL clusters the single meta-QTL model was most appropriate. Thus, the analysis identified nine meta-QTLs associated with slow wilting. Meta-analysis decreased the confidence intervals from an average of 21.4 cM for the eight QTL clusters to 10.8 cM for the meta-QTLs. Averaged R² values of the nine meta-QTLs in eight QTL clusters were 0.13 and ranged from 0.09 to 0.22. Meta-QTLs on Gm11 and Gm19 had the highest R² values (0.22 and 0.20, respectively).     

1-Substituted carbamoyl and thiocarbamoyl-4,5-dihydro-1H-pyrazoles as possible cytotoxic and antimicrobial agents

Two series of 1-substituted carbamoyl and thiocarbomoyl derivatives were prepared by either treating the corresponding pyrazole with the appropriate isocyanate and isothiocyanate respectively, or alternatively by condensing the appropriate diketone with the proper substituted semicarbazide or thiosemicarbazide. The structures of the prepared compounds were fully determined by analytical and spectral methods. Preliminary biological screening of the prepared compounds revealed significant antibacterial and cytotoxic activities for some compounds. Compounds 4a₂ and 4a₃ were found to be the most active against the human colon carcinoma HT29 (11.8 and 7.5?μg/mL, respectively) and human breast cancer MCF 7 (3.4 and 2.6?μg/mL, respectively) cell lines. The structure–activity relationship (SAR) and in silico drug relevant properties (HBD, HBA, tPSA, cLog P, molecular weight, % ABS, drug-likeness and drug score) further confirmed that the compounds are potential lead compounds for future drug discovery study.     

Thermoluminescent properties of some phosphor glasses based on aluminum oxide

The transparent thermoluminescent aluminum oxide‐based glass of 15Al₂O₃–35P₂O₅–25CaO–25Na₂CO₃, abbreviated as APCN (all in mol%) doped with different concentrations of SiO₂ from 0.0–500 ppm was prepared using a conventional melt‐quenching technique. The TL sensitivities of the prepared glasses were investigated at 3 Gy γ‐dose using a ⁶⁰Co source and measured at a heating rate 10 C/sec. The highest TL intensity of the material doped with SiO₂ was found at a concentration of 500 ppm (APCNSi₅). Deconvolution of the glow curve from APCNSi₅ resulted in four peaks at about 161, 194, 237 and 293 C with a Figure Of Merit (FOM) of 1.28%. The APCNSi₅ specimen had the best dosimetric properties when compared with the other samples. Reproducibility, repeatability, dose–response curve and fading effect were checked for peak 3, which appeared at about 237 C. The results displayed that the APCNSi₅ glass system was a low‐Z material (Z_eff ≈ 10), and had good reproducibility and good repeatability. Peak 3 showed good linearity over a dose range up to 20 Gy (R² = 0.999) and sublinearity behaviour was found. The signal from APCNSi₅ faded by about 11% after 2 days post irradiation, therefore it showed almost no significant loss. Such properties make the newly prepared glasses suitable for and highly recommended for use in γ‐dosimeters. Copyright © 2016 John Wiley & Sons, Ltd.     

Exome-Wide Association Analysis of Coronary Artery Disease in the Kingdom of Saudi Arabia Population

Coronary Artery Disease (CAD) remains the leading cause of mortality worldwide. Mortality rates associated with CAD have shown an exceptional increase particularly in fast developing economies like the Kingdom of Saudi Arabia (KSA). Over the past twenty years, CAD has become the leading cause of death in KSA and has reached epidemic proportions. This rise is undoubtedly caused by fast urbanization that is associated with a life-style that promotes CAD. However, the question remains whether genetics play a significant role and whether genetic susceptibility is increased in KSA compared to the well-studied Western European populations. Therefore, we performed an Exome-wide association study (EWAS) in 832 patients and 1,076 controls of Saudi Arabian origin to test whether population specific, strong genetic risk factors for CAD exist, or whether the polygenic risk score for known genetic risk factors for CAD, lipids, and Type 2 Diabetes show evidence for an enriched genetic burden. Our results do not show significant associations for a single genetic locus. However, the heritability estimate for CAD for this population was high (h² = 0.53, S.E. = 0.1, p = 4e^-12) and we observed a significant association of the polygenic risk score for CAD that demonstrates that the population of KSA, at least in part, shares the genetic risk associated to CAD in Western populations.     

Lentiviral vector-mediated overexpression of mutant ataxin-7 recapitulates SCA7 pathology and promotes accumulation of the FUS/TLS and MBNL1 RNA-binding proteins

Background

We used lentiviral vectors (LVs) to generate a new SCA7 animal model overexpressing a truncated mutant ataxin-7 (MUT ATXN7) fragment in the mouse cerebellum, in order to characterize the specific neuropathological and behavioral consequences of the genetic defect in this brain structure.

Results

LV-mediated overexpression of MUT ATXN7 into the cerebellum of C57/BL6 adult mice induced neuropathological features similar to that observed in patients, such as intranuclear aggregates in Purkinje cells (PC), loss of synaptic markers, neuroinflammation, and neuronal death. No neuropathological changes were observed when truncated wild-type ataxin-7 (WT ATXN7) was injected. Interestingly, the local delivery of LV-expressing mutant ataxin-7 (LV-MUT-ATXN7) into the cerebellum of wild-type mice also mediated the development of an ataxic phenotype at 8 to 12 weeks post-injection. Importantly, our data revealed abnormal levels of the FUS/TLS, MBNL1, and TDP-43 RNA-binding proteins in the cerebellum of the LV-MUT-ATXN7 injected mice. MUT ATXN7 overexpression induced an increase in the levels of the pathological phosphorylated TDP-43, and a decrease in the levels of soluble FUS/TLS, with both proteins accumulating within ATXN7-positive intranuclear inclusions. MBNL1 also co-aggregated with MUT ATXN7 in most PC nuclear inclusions. Interestingly, no MBNL2 aggregation was observed in cerebellar MUT ATXN7 aggregates. Immunohistochemical studies in postmortem tissue from SCA7 patients and SCA7 knock-in mice confirmed SCA7-induced nuclear accumulation of FUS/TLS and MBNL1, strongly suggesting that these proteins play a physiopathological role in SCA7.

Conclusions

This study validates a novel SCA7 mouse model based on lentiviral vectors, in which strong and sustained expression of MUT ATXN7 in the cerebellum was found sufficient to generate motor defects.

     

Microbial Regulation of Glucose Metabolism and Cell-Cycle Progression in Mammalian Colonocytes

A prodigious number of microbes inhabit the human body, especially in the lumen of the gastrointestinal (GI) tract, yet our knowledge of how they regulate metabolic pathways within our cells is rather limited. To investigate the role of microbiota in host energy metabolism, we analyzed ATP levels and AMPK phosphorylation in tissues isolated from germfree and conventionally-raised C57BL/6 mice. These experiments demonstrated that microbiota are required for energy homeostasis in the proximal colon to a greater extent than other segments of the GI tract that also harbor high densities of bacteria. This tissue-specific effect is consistent with colonocytes utilizing bacterially-produced butyrate as their primary energy source, whereas most other cell types utilize glucose. However, it was surprising that glucose did not compensate for butyrate deficiency. We measured a 3.5-fold increase in glucose uptake in germfree colonocytes. However, ¹³C-glucose metabolic-flux experiments and biochemical assays demonstrated that they shifted their glucose metabolism away from mitochondrial oxidation/CO₂ production and toward increased glycolysis/lactate production, which does not yield enough ATPs to compensate. The mechanism responsible for this metabolic shift is diminished pyruvate dehydrogenase (PDH) levels and activity. Consistent with perturbed PDH function, the addition of butyrate, but not glucose, to germfree colonocytes ex vivo stimulated oxidative metabolism. As a result of this energetic defect, germfree colonocytes exhibited a partial block in the G₁-to-S-phase transition that was rescued by a butyrate-fortified diet. These data reveal a mechanism by which microbiota regulate glucose utilization to influence energy homeostasis and cell-cycle progression of mammalian host cells.