Cloning and Expression of Aspergillus tamarii FS132 Lipase Gene in Pichia pastoris

A lipase gene (atl) was cloned from Aspergillus tamarii FS132 for the first time. The gene was found to have an open reading frame of 1024 base pairs (bp), and the coding region of the gene contained two introns (51 bp and 52 bp). Multi-alignment analysis of the deduced amino acid sequence indicated high homology between the enzyme and mono-and diacylglycerol lipases from fungi Aspergillus. The recombinant lipase was expressed in Pichia pastoris GS115 cells. The recombinant lipase was found to have a molecular mass of 36.7 kDa, and it exhibited lipase activity of 20 U/mL in culture supernatant when tributyrin was used as the substrate

Hearts from Mice Fed a Non-Obesogenic High-Fat Diet Exhibit Changes in Their Oxidative State, Calcium and Mitochondria in Parallel with Increased Susceptibility to Reperfusion Injury

High-fat diet with obesity-associated co-morbidities triggers cardiac remodeling and renders the heart more vulnerable to ischemia/reperfusion injury. However, the effect of high-fat diet without obesity and associated co-morbidities is presently unknown.To characterize a non-obese mouse model of high-fat diet, assess the vulnerability of hearts to reperfusion injury and to investigate cardiac cellular remodeling in relation to the mechanism(s) underlying reperfusion injury.Feeding C57BL/6J male mice high-fat diet for 20 weeks did not induce obesity, diabetes, cardiac hypertrophy, cardiac dysfunction, atherosclerosis or cardiac apoptosis. However, isolated perfused hearts from mice fed high-fat diet were more vulnerable to reperfusion injury than those from mice fed normal diet. In isolated cardiomyocytes, high-fat diet was associated with higher diastolic intracellular Ca2+ concentration and greater damage to isolated cardiomyocytes following simulated ischemia/reperfusion. High-fat diet was also associated with changes in mitochondrial morphology and expression of some related proteins but not mitochondrial respiration or reactive oxygen species turnover rates. Proteomics, western blot and high-performance liquid chromatography techniques revealed that high-fat diet led to less cardiac oxidative stress, higher catalase expression and significant changes in expression of putative components of the mitochondrial permeability transition pore (mPTP). Inhibition of the mPTP conferred relatively more cardio-protection in the high-fat fed mice compared to normal diet.This study shows for the first time that high-fat diet, independent of obesity-induced co-morbidities, triggers changes in cardiac oxidative state, calcium handling and mitochondria which are likely to be responsible for increased vulnerability to cardiac insults

Evaluation of Color and Spectral Behavior of a Novel Flowable Resin Composite after Water Aging: An In Vitro Study

Background: This study aimed to evaluate the color matching, light transmittance, and reflectance characteristics of the novel flowable resin composite OCF-001 (OCF). Methods: Fifty-four resin composite molds were made with simulated class I cavities of A2, A3, and A4 shades by filling the rubber mold interspace with Estelite Sigma Quick (ESQ), Gracefil Putty (GP) and Filtek Supremme Ultra (FSU). After applying the adhesive, three different flowable resin composites (n = 6), OCF, Gracefil LoFlo (GLF), and Supreme Ultra Flowable (SUF), were used to fill the cavities. A colorimeter was used to measure the color parameters (CIEDE2000). The color measurements were taken immediately and after 28 days. Data were analyzed using the nonparametric Kruskal-Wallis (alpha = 0.05) and Wilcoxon tests. The light transmittance and reflection characteristics were measured with a black background using a spectrophotometer under D65 illumination. Results: The Delta E-00, and Delta C of OCF was lower than other tested materials in A2 and A3 shades both immediately and after 28 days. OCF showed the highest transmittance characteristic, and a relatively stable reflectance curve in all the wavelengths. Conclusions: OCF showed better shade matching with the surrounding shades of A2 and A3, a relative uniform reflectance and higher light transmission properties

Influence of Novel Experimental Light-Cured Resin Cement on Microtensile Bond Strength

The purpose of this study was to evaluate the microtensile bond strength (mu TBS) and Knoop hardness number (KHN) of a novel experimental light-cured resin cement (HL). Eighteen flat dentin surfaces of human molars were polished using #600 SiC paper and bonded to CAD/CAM resin blocks with the respective resin cements and composites: HL, Panavia V5 (PV), and Clearfil AP-X (AP). All specimens were stored in distilled water at 37 degrees C for 24 h and 7 days. Scanning electron microscope (SEM) and energy dispersive X-ray (EDX) observations were performed to evaluate filler morphology and to detect the elements. The resin cements had a significant effect on the immediate mu TBS (F = 22.59, p < 0.05) and after water storage mu TBS (F = 22.83, p < 0.05). Significant differences (p < 0.05) in the KHN between the tested materials were observed, and HL indicated the highest KHN when compared with PV. HL showed a combination of the regular-shaped filler and spherical-shaped filler within the matrix. Silicon was detected in HL from the EDX evaluation. HL exhibited better bonding performance and polymerization, which may have contributed to the improvement of the adhesive strength

Preparation and anticoagulant activity of the phosphosulfomannan PI-88

A yeast-derived phosphomannan mixture was chemically sulfonated and the composition and structure of the product mixture was studied. This phosphosulfomannan mixture, PI-88, is currently under clinical evaluation as an anti-cancer agent. Analysis using capillary electrophoresis demonstrated that PI-88 was a multi-component mixture. Gel permeation chromatography provided four fractions of PI-88 that contained components which differed in size from disaccharide to hexasaccharide, and by degree of sulfation. These fractions were characterised by spectroscopic and chromatographic methods and the structure of PI-88 is that expected based On the structure of the phosphomannan starting material. The anticoagulant activity of these fractions was evaluated and the structural requirements for,activity are described. (C) 2002 Editions scientifiques et medicales Elsevier SAS. All rights reserved