Arg399Gln substitution in XRCC1 as a prognostic and predictive biomarker for prostate cancer: Evidence from 8662 subjects and a structural analysis

Abstract Background The Arg399Gln polymorphism in the X ray repair cross complementing group 1 gene ( XRCC1 ) may alter the risk of prostate cancer (PCa). The present study aimed to investigate the association of the XRCC1 Arg399Gln poly- morphism with PCa risk in an Iranian population, as followed by a meta analysis and an in silico analysis. Methods In a case control study, 360 subjects were included (180 men with PCa and 180 healthy controls). XRCC1 Arg399Gln genotyping was performed using the polymerase chain reaction restriction fragment length polymorphism method. In the meta analysis, 14 eligible studies were included to which our case control data were added to estimate the pooled odds ratios. Some bioinformatics tools were employed to evaluate the effects of Arg399Gln substitution on molecular aspects of the XRCC1 protein. Results Our case control study revealed a significant association between the XRCC1 Arg399Gln polymorphism and PCa risk. The data from overall meta analysis showed significant associations between the mentioned polymorphism and PCa risk in allelic and recessive genetic models. In addition, we observed statistically significant associations in stratified analyses by ethnicity, sample size and source of controls. Our in silico analysis showed that Arg399Gln substitution could be damaging with respect to the function and structure of the XRCC1 protein. Conclusions Based on these results, the XRCC1 Arg399Gln polymorphism might be a risk factor for PCa and it could be considered as a prognostic and predictive biomarker for susceptible men. KEYWORDS in silico analysis, meta analysis, prostate cancer, XRCC1 gen

Morphology of Rat Hippocampal CA1 Neurons Following Modified Two and Four-Vessels Global Ischemia Models

Background: An appropriate animal model of ischemia stroke is essential for evaluation of different therapeutic methods. Two and four-vessel global ischemia models are one of the most common types of transient cerebral ischemia. Objectives: In this study, the morphology of rat hippocampal CA1 neurons in modified models of two and four-vessel ischemia and reperfusion were evaluated. Materials and Methods: In this study, 20 Wistar rats were randomly divided into five groups. In group 2 and 3, both common carotid arteries were occluded for 10 minutes in either 3 or 24 hours of reperfusions, respectively. In group 4 and 5, both common carotid and vertebral arteries were occluded for 10 minutes in either 3 or 24 hours of reperfusions, respectively. Group 1 as control, underwent the whole surgery without any arteries occlusion. Hippocampi of the rats in all groups were processed and tissue sections were stained using the Nissl method. The morphology of CA1 neurons were studied under a light microscope and compared different groups. Results: In all groups ischemic changes were apparently observed in hippocampus CA1 neurons. In two-vessel occlusion model, after 3 and 24 hours of reperfusions, ischemic cells accounted for 14.9% and 23.2%, respectively. In four-vessel occlusion model, after 3 and 24 hours of reperfusions, ischemic cells accounted for 7.6% and 44.9% (P < 0.0001), respectively. Conclusions: Modified four-vessel occlusion model resulted in significant ischemic changes after 24 hours of reperfusion in CA1 neurons of rat hippocampus

Intranasal administration of endometrial mesenchymal stem cells as a suitable approach for Parkinson�s disease therapy

This study aimed to investigate the therapeutic effects of intranasal administration of human endometrium-derived stem cells (HEDSCs) in the mouse model of Parkinson�s disease (PD). Thirty days after intrastriatal injection of 6-OHDA, HEDSCs were administrated intranasally in three doses (104, 5 � 104 and 105 cells µl�1). During 120 days after stem cell administration, behavioral tests were examined. Then the mice were sacrificed and the fresh section of the substantia nigra pars compacta (SNpc) was used for detection of HEDSCs-GFP labeled by fluorescence microscopy method. In addition, immunohistochemistry was used to assay GFP, human neural Nestin, and tyrosine hydroxylase (TH) markers in the fixed brain tissue at the SNpc. Our data revealed that behavioral parameters were significantly improved after cell therapy. Fluorescence microscopy assay in fresh tissue and GFP analysis in fixed tissue were showed that the HEDSCs-GFP labeled migrated to SNpc. The data from immunohistochemistry revealed that the Nestin as a differential neuronal biomarker was expressed in SNpc. Also, TH as a dopaminergic neuron marker significantly increased after HEDSCs therapy in an optimized dose 5 � 104 cells µl�1. Our results suggest that intranasal administration of HEDSCs improve the PD symptoms in the mouse model of PD dose-dependent manner as a noninvasive method. © 2019, Springer Nature B.V

Reduction of truncated Kit Expression in Men with Abnormal Semen Parameters, Globozoospermia and History of Low or Fertilization Failure

Objective: Phospholipase C zeta 1 (PLCζ) is one of the main sperm factor involved in oocyte activation and other factors may assist this factor to induce successful fertilization. Microinjection of recombinant tr-kit, a truncated form of c-kit receptor, into metaphase II-arrested mouse oocytes initiate egg activation. Considering the potential roles of tr-KIT during spermiogenesis and fertilization, we aimed to assess expression of tr-KIT in sperm of men with normal and abnormal parameters and also in infertile men with previous failed fertilization and globozoospermia. Materials and Methods: This experimental study was conducted from September 2015 to July 2016 on 30 normozoospermic and 20 abnormozoospermic samples for experiment one, and also was carried out on 10 globozoospermic men, 10 men with a history low or failed fertilization and 13 fertile men for experiment two. Semen parameters and sperm DNA fragmentation were assessed according to WHO protocol, and TUNEL assay. Sperm tr-KIT was evaluated by flow cytometry, immunostaining and western blot. Results: The results show that tr-KIT mainly was detected in post-acrosomal, equatorial and tail regions. Percentage of tr-KIT-positive spermatozoa in abnormozoospermic men was significantly lower than normozoospermic men. Also significant correlations were observed between sperm tr-KIT with sperm count (r=0.8, P<0.001), motility (r=0.31, P=0.03) and abnormal morphology (r=-0.6, P<0.001). Expression of tr-KIT protein was significantly lower in infertile men with low/ failed fertilization and globozoospermia compared to fertile men. The significant correlation was also observed between tr-KIT protein with fertilization rate (r=-0.46, P=0.04). In addition, significant correlations were observed between sperm DNA fragmentation with fertilization rate (r=-0.56, P=0.019) and tr-KIT protein (r=-0.38, P=0.04). Conclusion: tr-KIT may play a direct or indirect role in fertilization. Therefore, to increase our insight regarding the role of tr-KIT in fertilization further research is warranted. © 2019 Royan Institute (ACECR). All rights reserved

Biodiesel production from jatropha seeds: Solvent extraction and in situ transesterification in a single step

The objective of this study was to investigate solvent extraction and in situ transesterification in a single step to allow direct production of biodiesel from jatropha seeds. Experiments were conducted using milled jatropha seeds, and n-hexane as extracting solvent. The influence of methanol to seed ratio (2:1–6:1), amount of alkali (KOH) catalyst (0.05–0.1 mol/L in methanol), stirring speed (700–900 rpm), temperature (40–60 °C) and reaction time (3–5 h) was examined to define optimum biodiesel yield and biodiesel quality after water washing and drying. When stirring speed, temperature and reaction time were fixed at 700 rpm, 60 °C and 4 h respectively, highest biodiesel yield (80% with a fatty acid methyl ester purity of 99.9%) and optimum biodiesel quality were obtained with a methanol to seed ratio of 6:1 and 0.075 mol/L KOH in methanol. Subsequently, the influence of stirring speed, temperature and reaction time on biodiesel yield and biodiesel quality was studied, by applying the randomized factorial experimental design with ANOVA (F-test at p = 0.05), and using the optimum values previously found for methanol to seed ratio and KOH catalyst level. Most experimental runs conducted at 50 °C resulted to high biodiesel yields, while stirring speed and reaction time did not give significantly effect. The highest biodiesel yield (87% with a fatty acid methyl ester purity of 99.7%) was obtained with a methanol to seed ratio of 6:1, KOH catalyst of 0.075 mol/L in methanol, a stirring speed of 800 rpm, a temperature of 50 °C, and a reaction time of 5 h. The effects of stirring speed, temperature and reaction time on biodiesel quality were not significant. Most of the biodiesel quality obtained in this study conformed to the Indonesian Biodiesel Standard

Retargeted adenoviruses for radiation-guided gene delivery

The combination of radiation with radiosensitizing gene delivery or oncolytic viruses promises to provide an advantage that could improve the therapeutic results for glioblastoma. X-rays can induce significant molecular changes in cancer cells. We isolated the GIRLRG peptide that binds to radiation-inducible 78 kDa glucose-regulated protein (GRP78), which is overexpressed on the plasma membranes of irradiated cancer cells and tumor-associated microvascular endothelial cells. The goal of our study was to improve tumor-specific adenovirus-mediated gene delivery by selectively targeting the adenovirus binding to this radiation-inducible protein. We employed an adenoviral fiber replacement approach to conduct a study of the targeting utility of GRP78-binding peptide. We have developed fiber-modified adenoviruses encoding the GRP78-binding peptide inserted into the fiber-fibritin. We have evaluated the reporter gene expression of fiber-modified adenoviruses in vitro using a panel of glioma cells and a human D54MG tumor xenograft model. The obtained results demonstrated that employment of the GRP78-binding peptide resulted in increased gene expression in irradiated tumors following infection with fiber-modified adenoviruses, compared with untreated tumor cells. These studies demonstrate the feasibility of adenoviral retargeting using the GRP78-binding peptide that selectively recognizes tumor cells responding to radiation treatment

Regiospecific analysis of Mono and Diglycerides in Glycerolysis products by GC x GC TOF-MS.

Comprehensive bidimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOF-MS) was used for the characterization of regiospecific mono- and diglycerides (MG-DG) content in the glycerolysis products derived from five different lipids included lard (LA), sun flower seed oil (SF), corn oil (CO), butter (BU), and palm oil (PA). The combination of fast and high temperature non-orthogonal column set namely DB17ht (6 m × 0.10 mm × 0.10 μm) as the primary column and SLB-5 ms (60 cm × 0.10 mm × 0.10 μm) as the secondary column was applied in this work. System configuration involved high oven ramp temperature to obtain precise mass spectral identification and highest effluent’s resolution. 3-Monopalmitoyl-sn-glycerol (MG 3-C16) was the highest concentration in LA, BU and PA while monostearoyl-sn-glycerol (MG C18) in CO and 1,3-dilinoleol-rac-glycerol (DG C18:2c) in SF. Principal component analysis accounted 82% of variance using combination of PC1 and PC2. The presence of monostearoyl-sn-glycerol (MG C18), 3-Monopalmitoyl-sn-glycerol (MG 3-C16), 1,3-dilinoleol-rac-glycerol (DG C18:2c), 1,3-dipalmitoyl-glycerol (DG 1,3-C16), and 1,3-dielaidin (DG C18:1t) caused differentiation of the samples tested

Direct enzymatic esterification of cotton and Avicel with wild-type and engineered cutinases

In this work, the surface of cellulose, either Avicel or cotton fabric, was modified using cutinases without any previous treatment to swell or to solubilise the polymer. Aiming further improvement of cutinase ester synthase activity on cellulose, an engineered cutinase was investigated. Wild-type cutinase from Fusarium solani and its fusion with the carbohydrate-binding module N1 from Cellulomonas fimi were able to esterify the hydroxyl groups of cellulose with distinct efficiencies depending on the acid substrate/solvent system used, as shown by titration and by ATR-FTIR. The carbonyl stretching peak area increased significantly after enzymatic treatment during 72 h at 30 °C. Cutinase treatment resulted in relative increases of 31 and 9 % when octanoic acid and vegetable oil were used as substrates, respectively. Cutinase-N1 treatment resulted in relative increases of 11 and 29 % in the peak area when octanoic acid and vegetable oil were used as substrates, respectively. The production and application of cutinase fused with the domain N1 as a cellulose ester synthase, here reported for the first time, is therefore an interesting strategy to pursuit.This work was co-funded by the European Social Fund through the management authority POPH and FCT, Postdoctoral fellowship reference: SFRH/BPD/47555/2008. The authors also want to thank Doctor Raul Machado for his valuable help on FTIR spectral data treatment