The Effect of Continued Training with Crocin on Apoptosis Markers in Liver Tissue of High Fat Diet Induced Diabetic Rats

Diabetes mellitus (DM) disease can affect process of apoptosis byincreasing oxidative stress, nevertheless exercise and crocin can improve apoptosis; therefore present study aimed to investigate the effect of continued trainingwith crocin on apoptosis markers in liver tissue of diabetic rats. In this experimental study 32 diabetic rats based on fasting glucose divided into four groups ofeight rats including: 1) sham, 2) training, 3) crocin, and 4) training with crocinalso for investigate the effect of DM induction on apoptosis markers, eight healthyrats assigned in healthy control group. During eight weeks groups 2 and 4 ran60 minutes on treadmill with intensity of 50–55% maximum speed for three sessions per week and groups 3 and 4 received 25 mg/kg/day crocin peritoneally.Shapiro–Wilk, one-way ANOVA with Tukey’s post-hot tests were used for statistical analysis of data (P ≤ 0.05). DM induction significantly increased Bcl-2 aswell as decreased Bax and P52 (P ≤ 0.05) nevertheless training and training withcrocin significantly decreased Bcl-2 and increased Bax and P53 (P ≤ 0.05); crocinsignificantly decreased Bcl-2 and increased P53 (P ≤ 0.05) and training with crocin had higher effect on increase of Bax and P53 compare to training (P ≤ 0.05)also increase of Bax compare to crocin. Although training and crocin alone canimprove apoptotic markers in diabetic rats, nevertheless training simultaneouslywith crocin have better effects than training alone.     

Noninvasive Fetal Sex Determination by Real-Time PCR and TaqMan Probes

Background:Noninvasive fetal sex determination by analyzing Y chromosome-specific sequences is very useful in the management of cases related to sex-linked genetic diseases. The aim of this study was to establish a non-invasive fetal sex determination test using Real-Time PCR and specific probes.Methods:The study was a prospective observational cohort study conducted from August 2018 to September 2019. Venous blood samples were collected from 25 Iranian pregnant women at weeks 7 to 25 of gestation. Cell-free DNA (cfDNA) was isolated from the plasma of samples and fetal sex was determined by SRY gene analysis using the Real-Time PCR technique. In the absence of SRY detection, the presence of fetal DNA was investigated using cfDNA treated with BstUI enzyme and PCR for the epigenetic marker RASSF1A.Results:Of the total samples analyzed, 48% were male and 52% female. The RASSF1A assay performed on SRY negative cases also confirmed the presence of cell-free fetal DNA. Genotype results were in full agreement with neonate gender, and the accuracy of noninvasive fetal sex determination was 100%.Conclusion:Fetal sex determination using the strategy applied in this study is noninvasive and highly accurate and can be exploited in the management of sex-linked genetic diseases.Key Words: Cell-free fetal DNA, Fetal sex determination, Noninvasive prenatal diagnosis, Sex-linked genetic diseases, SRY     

Influence of nitric oxide in protection of tomato seedling against oxidative stress induced by osmotic stress

Osmotic stress associated with drought and salinity is a serious problem that inhibits the growth of plants mainly due to disturbance of the balance between production of ROS and antioxidant defense and causes oxidative stress. In this research, sodium nitroprusside (SNP) was used as NO donor in control and drought-stressed plants, and the role of NO in reduction of oxidative damages were investigated. In this study, we observed that SNP pretreatment prevented drought-induced decrease in RWC and membrane stability index, increase in lipid peroxidation and lipoxygenase activity and increase in hydrogen peroxide content. However, pretreatment of plants with SNP and phenyl 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (a NO scavenger) reversed the protective effects of SNP suggesting that protective effect by SNP is attributable to NO release. In addition, the relationship between these defense mechanisms and activity of antioxidant enzymes were checked. Results showed that in drought-stressed plants ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and catalase activities were elevated over the controls, while GR decreased under drought condition. Activity of GPX was inhibited under SNP pretreatment in drought-stressed plants specially, while the activity of APX and GR increased under SNP pretreatment and it seems that under this condition APX had a key role of detoxification of ROS in tomato plants. This result corresponded well with ASA and total acid-soluble thiols content. Therefore, reduction of drought-induced oxidative damages by NO in tomato leaves is most likely mediated through either NO ability to scavenge active oxygen species or stimulation of antioxidant enzyme such as APX.     

Comparison of different artificial neural network architectures in modeling of Chlorella sp. flocculation

Biodiesel production from microalgae feedstock should be performed after growth and harvesting of the cells, and the most feasible method for harvesting and dewatering of microalgae is flocculation. Flocculation modeling can be used for evaluation and prediction of its performance under different affective parameters. However, the modeling of flocculation in microalgae is not simple and has not performed yet, under all experimental conditions, mostly due to different behaviors of microalgae cells during the process under different flocculation conditions. In the current study, the modeling of microalgae flocculation is studied with different neural network architectures. Microalgae species, Chlorella sp., was flocculated with ferric chloride under different conditions and then the experimental data modeled using artificial neural network. Neural network architectures of multilayer perceptron (MLP) and radial basis function architectures, failed to predict the targets successfully, though, modeling was effective with ensemble architecture of MLP networks. Comparison between the performances of the ensemble and each individual network explains the ability of the ensemble architecture in microalgae flocculation modeling.     

Concomitant Up-Regulation of Hsa- Mir-374 and Down-Regulation of Its Targets,GSK-3β and APC,in Tissue Samples of Colorectal Cancer

Background:The WNT-pathway is involved in several cancers, including colorectal cancer (CRC). Many cell signaling components and pathways are controlled by microRNAs. The main purpose of the present study was to investigate the expression of hsa-miR-374, and its two target genes of the Wnt-pathway in CRC clinical samples.Methods:In this study, we predicted the miRNAs targeting key genes of WNT-pathway using bioinformatics algorithms. The expression levels of hsa-miR-374, APC and GSK-3β on 48 pairs of Formalin-Fixed Paraffin-Embedded (FFPE) CRC tumors and marginal-tumors were evaluated using real time-PCR. Additionally, the hsa-miR-374a-5p precursor sequence was amplified by whole-blood DNA as a template. This amplicon was cloned into pEGFP-c1 expression vector and transfected into SW742 cells. Aside from this, MTT assay was performed to evaluate the effect of miR-374 on cell viability. Results:The bioinformatics analysis indicated that hsa-miR-374 binds to the regulatory region the key components of WNT-pathway, including APC and GSK-3β considering the recognition elements and mirSVR scores. Our results revealed significant down-regulation of GSK-3β (0.94 times, p= 0.0098) and APC (0.96 times, p= 0.03) and up-regulation of miR-374 (1.22 times, p= 0.0071) on tumor samples compared with their normal pairs. Meanwhile, the results of the over-expression of miR-374 showed down-regulation of APC and GSK-3β. MTT-assay also indicated that the miR-374 increased cell survival.Conclusion:The results of our study indicated a concomitant change in the expression of miR-374 and its two related target genes, in clinical samples of CRC. Hsa-miR-374 might be as a helpful biomarker or therapeutic target in CRC.Key Words: Colorectal cancer, GSK-3β, miR-374, WNT     

Effects of Chromium and Carnitine Co-supplementation on Body Weight and Metabolic Profiles in Overweight and Obese Women with Polycystic Ovary Syndrome: a Randomized,Double-Blind,Placebo-Controlled Trial

The primary aim of our study was to determine the influence of taking chromium plus carnitine on insulin resistance, with a secondary objective of evaluating the influences on lipid profiles and weight loss in overweight subjects with polycystic ovary syndrome (PCOS). In a 12-week randomized, double-blind, placebo-controlled clinical trial, 54 overweight women were randomly assigned to receive either supplements (200 μg/day chromium picolinate plus 1000 mg/day carnitine) or placebo (27/each group). Chromium and carnitine co-supplementation decreased weight (− 3.6 ± 1.8 vs. − 1.0 ± 0.7 kg, P < 0.001), BMI (− 1.3 ± 0.7 vs. − 0.3 ± 0.3 kg/m², P < 0.001), fasting plasma glucose (FPG) (− 5.1 ± 6.0 vs. − 1.1 ± 4.9 mg/dL, P = 0.01), insulin (− 2.0 ± 1.4 vs. − 0.2 ± 1.2 μIU/mL, P < 0.001), insulin resistance (− 0.5 ± 0.4 vs. − 0.04 ± 0.3, P < 0.001), triglycerides (− 18.0 ± 25.2 vs. + 5.5 ± 14.4 mg/dL, P < 0.001), total (− 17.0 ± 20.3 vs. + 3.6 ± 12.0 mg/dL, P < 0.001), and LDL cholesterol (− 13.3 ± 19.2 vs. + 1.4 ± 13.3 mg/dL, P = 0.002), and elevated insulin sensitivity (+ 0.007 ± 0.005 vs. + 0.002 ± 0.005, P < 0.001). In addition, co-supplementation upregulated peroxisome proliferator-activated receptor gamma (P = 0.02) and low-density lipoprotein receptor expression (P = 0.02). Overall, chromium and carnitine co-supplementation for 12 weeks to overweight women with PCOS had beneficial effects on body weight, glycemic control, lipid profiles except HDL cholesterol levels, and gene expression of PPAR-γ and LDLR. Clinical trial registration number: http://www.irct.ir: IRCT20170513033941N38.

     

Genetically engineered phage harbouring the lethal catabolite gene activator protein gene with an inducer-independent promoter for biocontrol of Escherichia coli

Complications of chemotherapy, such as appearance of multidrug resistance, have persuaded researchers to consider phage therapy as a new method to combat bacterial infections. In vitro experiments were performed to assess the therapeutic value of genetically modified phages for controlling gastrointestinal Escherichia coli O157:H7 cells in Luria–Bertani (LB) media and contaminated cow milk. We constructed a modified nonreplicating M13-derived phage expressing a lethal catabolite gene activator protein (CAP) that is a Glu181Gln mutant of CAP. The modified phagemid was propagated in the lethal CAP-resistant strain XA3DII. Time–kill assay experiments showed a considerable reduction in the number of surviving bacteria in both LB media and contaminated cow milk. Our further study using other test strains demonstrated that the host range of lethal phage is limited to E. coli strains that produce pili. This study provides a possible strategy for the exploitation of genetically engineered nonlytic phages as bactericidal agents by minimizing the risk of release of progeny phages and endotoxins into the environment. The phage was engineered to remain lethal to its bacterial target, but incapable of replicating therein. Furthermore, the addition of an inducer to express the lethal protein is not required.     

Donor/recipient interactions affecting plasmid transfer amongStreptomyces species: A conjugative plasmid will mobilize nontransferable plasmids in soil

Streptomyces parvulus was used as the recipient for plasmid pIJ303 and pIJ211, two conjugative plasmids derived from the self-transmissible plasmid pIJ101. One of the resulting transconjugantS. parvulus strains containing plasmid pIJ303 was used withS. lividans to evaluate the effects of the host strain on the frequency of pIJ303 transfer betweenStreptomyces species. Only 30% ofS. parvulus cells acquired plasmid pIJ303 in crosses in whichS. lividans was the donor, whereas 100% ofS. lividans cells acquired the plasmid whenS. parvulus was the donor. This indicates that the frequency of transfer of the conjugative plasmid was determined by the recipient. The other resulting transconjugantS. parvulus strain containing plasmid pIJ211 was evaluated for its ability to mobilize the nonconjugative plasmid pIJ702 fromS. lividans, on agar and in sterile soil. AfterS. lividans containing pIJ702 was crossed on agar and in sterile soil withS. parvulus containing pIJ211, recombinantS. parvulus colonies carrying pIJ702 and expressing pigments characteristic of both species were recovered, from both agar and soil. Although a large percentage ofS. parvulus transconjugants lost pIJ211 during incubation in soil, the mobilization of pIJ702 fromS. lividans intoS. parvulus still occurred. Plasmid integration into the chromosome of the donor and the transconjugant was evaluated by Southern blot hybridization. Hybridization of plasmid pIJ303, with chromosomal DNA fromS. lividans andS. parvulus transconjugants, using biotinylated DNA, indicated that no integration had occurred. Genetic exchange betweenStreptomyces species also occurred in a liquid medium. The finding of plasmid mobilization in soil is significant. It demonstrates that genetic exchange in the environment can occur between released genetically engineeredStreptomyces species and nativeStreptomyces species that contain conjugative plasmids.Paper of the Idaho Agricultural Experiment Station.     

Metabolic flexibility determines human NK cell functional fate in the tumor microenvironment

1. Download : Download high-res image (348KB)
2. Download : Download full-size image