Review of Molecular Techniques for the Identification of Bacterial Communities in Biological Effluent Treatment Facilities at Pulp and Paper Mills

Cholesteryl ester transfer protein (CETP) activity was measured in d > 1.21 g/ml plasma from hypertriglyceridemic patients and compared with normolipidemic subjects. The assay consisted in measuring the specific transfer of [3H]cholesteryl oleate from a prelabelled, apo E-poor HDL fraction to VLDL after incubation at 37 degrees C in the presence of the d > 1.21 g/ml plasma sample: the lipoproteins were then separated by precipitation with dextran sulfate/Mg2+ solution. Increasing the volume of d > 1.21 g/ml plasma or purified human CETP in the assay produced linear responses in measured activity, whereas, either during incubation at 4 degrees C or in the presence of rat plasma instead of human plasma, the transfer of [3H]cholesteryl oleate to VLDL was not stimulated. Thus, the assay reflects changes in CETP in the sample and appears to be suitable for measuring CETP activity in d > 1.21 g/ml plasma. CETP activity was very similar in the two groups of normolipidemic subjects considered: adolescents (203 +/- 11 nmol esterified cholesterol transferred per 8 h/ml plasma) and adults (215 +/- 5). Patients were grouped into lipoprotein-lipase (LPL)-deficient and non-LPL-deficient according to their enzyme activity in postheparin plasma. CETP activity was highly increased in LPL-deficient, severe hyperchylomicronemic patients (430 +/- 42) and was directly correlated with VLDL levels in the non-LPL-deficient individuals. Marked differences were observed in the lipid composition of HDL and apolipoprotein A-I levels among patients and controls. In the control group, CETP activity was correlated only with HDL-triglyceride and HDL-triglyceride/apo A-I mass ratio, which is compatible with the physiological role of CETP in transferring triglyceride to HDL from other lipoprotein particles. When all hypertriglyceridemic patients were considered together, CETP activity was inversely correlated with apo A-I and HDL-cholesterol, whereas it was directly correlated with HDL-triglyceride/HDL-cholesterol and HDL-triglyceride/apo A-I mass ratios. The results indicate that the enhanced CETP activity associated with hypertriglyceridemia contributes to the compositional change of HDL, which in turn may be responsible for the reduction of HDL levels in this condition.

Functional genomics approaches are widely implemented in current research and have found application in many areas of biology. This review will present research fields, novel findings and new tools developed in the cytochrome P450 field using the functional genomics techniques. The most widely used method is microarray technology, which has already greatly contributed to the understanding of the cytochromes P450 function and expression. Several focused CYP microarrays have been developed for genotyping, toxicogenomics and studies of CYP function of many different organisms. Our contribution to the CYP field by development of Steroltalk microarrays to study the cross-talk of cholesterol homeostasis and drug metabolism is also presented.

Kluyveromyces marxianus is a yeast that can be used as a microbial factory. However, little is known about its response to stress conditions. This work evaluated the response of this yeast against ethanol, acetic acid, isoamyl alcohol, and hydrogen peroxide as stress agents. Cytotoxicity assays were performed to assess the residual viability using a direct method (CFU counting) and an indirect method based on the reduction in MTT. Then, fermentation kinetics were performed at IC30 and IC50 for each stress factor to evaluate the effect of moderate and intense stress. This work is the first report presenting IC50 values for ethanol (21.82 g/L), acetic acid (1.19 g/L), isoamyl alcohol (2.74 g/L), and hydrogen peroxide (0.09 g/L) in K. marxianus. The IC50 values for the indirect method are between 3.7 and 68% higher than those for the direct method. Hydrogen peroxide and ethanol were the stress agents showing the highest overestimations. The results presented here demonstrated the overestimation of cell viability by the indirect method. Direct CFU counting is an adequate method to determine yeast viability during toxicity studies of chemical compounds. It was also established that ethanol and hydrogen peroxide have the highest toxicity against K. marxianus ITD-01005 during fermentation at concentrations equivalent to IC30 and IC50 of each stress agent.

All traditional mezcal producers use artisan methods to produce mezcal. The low technological development in the elaboration processes results in low yield and high residual sugar concentration. First, this work optimized the concentration of initial sugars and yeast-assimilable nitrogen (YAN) in Agave durangensis juice fermentation at the laboratory level. A yield near 0.49 g EtOH/g sugar and a productivity of 1.54 g EtOH/L*h was obtained with an initial sugar concentration of 120 g/L and a YAN concentration of 0.227 g/L. Only Saccharomyces cerevisiae was found after 24 h of incubation at laboratory level, using MALDI-TOF identification. Agave durangensis heads crushed by the artisan process were used to test the inoculant performance. A mezcal yield of 11.6 kg agave/L of mezcal was obtained using the S cerevisiae inoculant and nitrogen addition, which was significantly different (p < 0.05) from other treatments. The population dynamics during fermentation were analyzed through isolation and identification using MALDI-TOF. Several yeast species (Pichia kluyveri, Torulaspora delbrueckii, Zygosaccharomyces bailii, and Saccharomyces cerevisiae) were found at the beginning of fermentation. Nonetheless, only S. cerevisiae was found at the end of fermentation. The implantation of the inoculant used was confirmed through the comparative analysis of amplification patterns of the GTG5 microsatellite of the strains identified as S. cerevisiae, finding that the inoculated strain proportion was greater than 80% of the yeast population. A technological alternative to increase the efficiency of the process is combining the addition of YAN and the inoculation of the native S. cerevisiae, which was isolated from artisan alcoholic fermentation of agave to produce mezcal.