Anatomical basis for cell transplantation into mouse seminiferous tubules

Cell transplantation into the seminiferous tubules is a useful technique for the study of physiological and pathological conditions affecting the testis. However, the precise three-dimensional organization and, particularly, the complex connectivity of the seminiferous network have not yet been thoroughly characterized. To date, the technical approaches to address these issues have included manual dissection under the stereomicroscope, reconstruction of histological serial sections, and injection of contrast dyes, but all of them have yielded only partial information. Here, using an approach based on the microinjection of a self-polymerizing resin followed by chemical digestion of the surrounding soft tissues, we reveal fine details of the seminiferous tubule scaffold and its connections. These replicas of the testis seminiferous network were studied by scanning electron microscopy. The present results not only establish a morphological basis for more precise microinjection into the mouse seminiferous tubules but also enable a more profound investigation of physiological and embryological features of the testis.     

Immobilization of β-Glucosidase and Its Application for Enhancement of Aroma Precursors in Muscat Wine

Enzyme immobilization is becoming more widely practised in biotechnology because of the advantages that this method brings. In this study, commercial β-glucosidase for aroma released in winemaking was immobilized on diverse supports (alginate–chitin, chitosan–chitin) by using different methods. It was found that the most appropriate matrix was chitosan by adsorption and reticulation. The optimal immobilization conditions were pH 3.5, immobilization time 120 min, and concentration of cross-linker glutaraldehyde 0.25 %. Stability of the immobilized enzymes was assessed, which revealed a number of advantages, such as a lower enzyme dose required for immobilization (367 times lower than the free enzyme dose recommended by the manufacturer), high stability over time, and reusability. In vitro studies of cellobiose and in vivo studies of wine and aroma precursors isolated from grape must yielded similar outcomes with respect to enzyme hydrolysis of free and immobilized proteins.     

Effects of Reagent Concentrations on Advanced Oxidation of Amoxicillin by Photo-Fenton Treatment

The degradation and mineralization of amoxicillin in an aqueous solution was accomplished by using a photo-Fenton treatment. An ultraviolet light source with a 254-nm wavelength was used with hydrogen peroxide (H2O2) and iron(II). The effects of reagent concentrations on amoxicillin degradation and mineralization were investigated systematically by using the Box-Behnken statistical experiment design. Amoxicillin (10–200??mgL-1), H2O2 (10–500??mgL-1), and iron(II) (0–50??mgL-1) concentrations were considered independent variables; the percent amoxicillin degradation and the total organic carbon (TOC) removal (mineralization) were the objective functions to be maximized. Both H2O2 and iron(II) concentrations affected the extent of the amoxicillin degradation and mineralization. The amoxicillin degradation was completed within 2.5?min, and 53% mineralization took place within 60?min. The optimum H2O2∶Fe∶amoxicillin ratio that resulted in complete amoxicillin degradation and 53% mineralization was 100∶40∶105??mgL-1.     

Esterification of Free Fatty Acids Using Layered Carboxylates and Hydroxide Salts as Catalysts

Layered lanthanum and copper(II) laurates were tested as catalysts for methanolic esterification of lauric acid (C₁₂). Factorial design evidenced a positive effect of reaction temperature and amount of catalyst on ester conversion, while the acid:alcohol molar ratio (MR) had a negative effect. With lanthanum laurate, 90.5% conversion was reached at 140???C with 10% catalyst and MR of 6:1. Although the highest conversion with copper(II) laurate converted 81.5% under the same conditions, results were better with a lower amount of catalyst (78% ester at 140???C with 2% catalyst and MR of 6:1). A calcium hydroxide succinate was also investigated showing better catalytic performance than the well-known CaO, under mild reaction conditions (120???C with 1% catalyst and MR of 3:1). Although layered carboxylates can be successively reused, the layered calcium hydroxide salt transformed into inactive CaCO₃ in the second reuse reaction.     

gef Gene Expression in MCF-7 Breast Cancer Cells is Associated with a Better Prognosis and Induction of Apoptosis by p53-Mediated Signaling Pathway

Breast cancer research has developed rapidly in the past few decades, leading to longer survival times for patients and opening up the possibility of developing curative treatments for advanced breast cancer. Our increasing knowledge of the biological pathways associated with the progression and development of breast cancer, alongside the failure of conventional treatments, has prompted us to explore gene therapy as an alternative therapeutic strategy. We previously reported that gef gene from E. coli has shown considerable cytotoxic effects in breast cancer cells. However, its action mechanism has not been elucidated. Indirect immunofluorescence technique using flow cytometry and immunocytochemical analysis were used to detect breast cancer markers: estrogen (ER) and progesterone (PR) hormonal receptors, human epidermal growth factor receptor-2 proto-oncogene (c-erbB-2), ki-67 antigen and p53 protein. gef gene induces an increase in ER and PR expressions and a decrease in ki-67 and c-erbB-2 gene expressions, indicating a better prognosis and response to treatment and a longer disease-free interval and survival. It also increased p53 expression, suggesting that gef-induced apoptosis is regulated by a p53-mediated signaling pathway. These findings support the hypothesis that the gef gene offers a new approach to gene therapy in breast cancer.     

Insights into the Adhesive Mechanisms of Tree Frogs using Artificial Mimics

Elastic, microstructured surfaces (hydrophobic and hydrophilic) mimicking the surface structure of tree‐frog toe‐pads are fabricated. Their adhesion and friction behaviour in the presence of a liquid layer is evaluated and compared to flat controls. Tree‐frog‐like patterns are beneficial for wet adhesion only if the liquid does not wet the surface. The situation is different in friction, where the surface structure lead to significantly higher friction forces only if the liquid does wet the surface. Taking into account that tree‐frog attachment pads are hydrophilic and that their secretion wets all kind of surfaces, our results indicate that the surface structure in tree‐frog toe‐pads has been developed for climbing, when shear (friction) forces are involved. These results evidence the benefits and limitations of the surface design (microstructure and hydrophilicity) for adhesion and friction under wet conditions.     

Thermal,mechanical and water adsorption properties of corn starch–carboxymethylcellulose/methylcellulose biodegradable films

The objective of this study was to investigate the effect of the addition of methylcellulose and carboxymethylcellulose on the thermal, mechanical and water adsorption properties of starch-based films plasticized with glycerol or polyethylene glycol (PEG). Mechanical tests showed that as the methylcellulose and carboxymethylcellulose proportion increased, starch films became more resistant to break, resulting in higher TS values. Besides there has been a positive effect on the elasticity of starch films realized by a considerable increase in E% values. Depending on the plasticizer type, either single or dual glass transitions were seen in DSC thermograms. One glass transition temperature was observed for films plasticized with glycerol, on the contrary, dual glass transitions were detected for PEG plasticized films. This behavior was attributed to the phase separation of the PEG. In addition, the presence of an endothermic peak in the thermograms of PEG plasticized films was taken as another indicator of the phase separation. As a result, it was suggested that PEG was not as compatible as glycerol with the composite polysaccharide matrix and plasticizer type was the main factor that shaped the thermal profiles of the film samples. Water adsorption isotherm data showed that samples displayed nonlinear sorption profile which is typical for hydrophilic films. In all films tested, equilibrium moisture contents, increased almost linearly up to a a_w of 0.65–0.85, beyond where a sharp increase was noted. Adsorption data was adequately fitted by BET and GAB models. Eventually, it can be concluded that film forming properties of starch can be improved by incorporation of methylcellulose and carboxymethylcellulose to the polymer matrix.     

Modulatory effects of yerba maté (Ilex paraguariensis) on the PI3K‐AKT signaling pathway

The aim of this study was to evaluate the effects of yerba maté (YM) extract on the phosphatidylinositol 3‐kinase (PI3K)‐AKT signaling pathway in vivo. The mice were introduced to either standard‐ or high‐fat diet (HFD). After 8 weeks on an HFD, mice were randomly assigned to one of the two treatment conditions, water or yerba maté extract at 1.0 g/kg. After treatment, glucose blood level and hepatic insulin response were evaluated. Liver tissue was examined to determine the mRNA levels using the PI3K‐AKT PCR array. The nuclear translocation of forkhead box O1 (FOXO1) was determined by an electrophoretic mobility‐shift assay. Our data demonstrated that yerba maté extract significantly decreased the final body weight, glucose blood levels, and insulin resistance of mice. Molecular analysis demonstrated that an HFD downregulated Akt2, Irs1, Irs2, Pi3kca, Pi3kcg, and Pdk1; after yerba maté treatment, the levels of those genes returned to baseline. In addition, an HFD upregulated Pepck and G6pc and increased FOXO1 nuclear translocation. The intervention downregulated these genes by decreasing FOXO1 nuclear translocation. The results obtained demonstrate for the first time the specific action of yerba maté on the PI3K‐AKT pathway, which contributed to the observed improvement in hepatic insulin signaling.