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

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

Kinetics of Transesterification of Soybean Oil

Transesterification of soybean oil with methanol was investigated. Three stepwise and reversible reacttons are believed to occur. The effect of variations in mixing intensity (Reynolds number = 3,100 to 12,400) and temperature (30 to 70\u27CJ on the rate oireaction were studied while the molar ratio of alcohol to triglycerol (6:l) and the concentration of catalyst (0.20 wt% based on soybean oil) were held constant. The variations in mixing intensity appear to effect the reaction parallel to the variations in temperature. A reaction mechanism consisting of an initial mass transfer-controlled region followed by a kinetically controlled region is proposed. The experimental data for the latter region appear to be a good fit into a second-order kinetic mechanism. The reaction rate constants and the activation energies were determined for all the forward and reverse reactions

Glycerolysis of Fats and Methyl Esters.

The glycerolysis of methyl esters and triglycerides with crude glycerol. a coproduct from the transesterification of triglycerides, was studied. Three procedures were followed ior this conversion. The first procedure was a one-step glycerolysis with methyl esters. The second procedure was a two-step process. This proced~~irnev olved an initial partial glycerolysis with methyl esters, followed by fat glycerolysis. The third procedure u,as a simultaneous glycerolysis n,ith methyl esters and triglycerides. In the glycerolysis with methyl esters, the removal of methanol is vital to the production of mono- and diglycerides. Methanol was removed either by drawing vacuum on the reactor or by stripping methanol out by means of an inert carrier gas (nitrogen]. Different molar ratios of methyl esters to glycerol were tested in the first two processes. At low concentration of methyi esters, total conversion oi methyl esters to mono- and diglycerides was achieved. As the concentration oi methyl esters was increased, the conversion of methyl esters to mono and diglvcerides was decreased. Furthermore, the ratio of mono- to diglycerides was also higher at lower roncentrations of methyl esters. The conversion of triglycerides in the two-step process with crude glycerol was similar to a one-step fat glycerolysis with pure glycerol. The composition of different coniponents and the ratio of mono to diglycerides were also comparabl

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