Characterization of Effluents Generated During the Cleaning of Expansion Tanks Used to Store Raw Milk in Brazil / Caracterização dos Efluentes Gerados Durante a Limpeza dos Tanques de Expansão Usados para Armazenar Leite Cru no Brasil

The objective of this study was to characterize the effluents generated during the cleaning of the expansion tanks used to store raw milk in Brazil. Fifteen expansion tanks were chosen and three samples collected from each tank, as well as measuring the temperature and volume. The biological oxygen demand (BOD), chemical oxygen demand (COD), acidity, pH, alkalinity, total phosphorus, nitrogen, fats and oils, hardness, turbidity and total dissolved solids were determined. The statistical design adopted was completely at random and the Scott-Knott test was adopted to compare the means that differed significantly according to the F test. The multivariate data analysis was also carried out and studied by the principal component analysis (PCA) and the grouping analysis for similarity, as represented by a dendogram. The effluents were not treated and some of them (26.7%) were discharged into water courses. The concentrations of the organic matter indicators and the results of the physicochemical analyses of the expansion tank washing effluents showed that the majority of the variables were above the legal standards established by Brazilian legislation. Thus an adequate destiny for these effluents is required. The PCA showed that the variables with the greatest influence on the characteristics of the expansion tank effluents were related to the COD and nitrogen concentrations. A total of 88% of similarity was found between the effluents of the 15 tanks. According to the concentrations obtained for the phosphorus, nitrogen, COD and BOD of the effluents, they could be treated by the wetlands systems.The objective of this study was to characterize the effluents generated during the cleaning of the expansion tanks used to store raw milk in Brazil. Fifteen expansion tanks were chosen and three samples collected from each tank, as well as measuring the temperature and volume. The biological oxygen demand (BOD), chemical oxygen demand (COD), acidity, pH, alkalinity, total phosphorus, nitrogen, fats and oils, hardness, turbidity and total dissolved solids were determined. The statistical design adopted was completely at random and the Scott-Knott test was adopted to compare the means that differed significantly according to the F test. The multivariate data analysis was also carried out and studied by the principal component analysis (PCA) and the grouping analysis for similarity, as represented by a dendogram. The effluents were not treated and some of them (26.7%) were discharged into water courses. The concentrations of the organic matter indicators and the results of the physicochemical analyses of the expansion tank washing effluents showed that the majority of the variables were above the legal standards established by Brazilian legislation. Thus an adequate destiny for these effluents is required. The PCA showed that the variables with the greatest influence on the characteristics of the expansion tank effluents were related to the COD and nitrogen concentrations. A total of 88% of similarity was found between the effluents of the 15 tanks. According to the concentrations obtained for the phosphorus, nitrogen, COD and BOD of the effluents, they could be treated by the wetlands systems

Differencial proteome of clear-cell renal cell carcinoma (ccRCC) tissues

Purpose We attempted to detect, for the first time in a Brazilian cohort, differences in protein expression between clear-cell renal cell carcinoma (ccRCC) and their normal adjacent tissues, aiming to identify biomarkers and/or therapeutic target candidates for this disease. Material and Methods Twenty-four ccRCC and adjacent normal tissues were collected after surgery and their protein extracts were quantified, pooled and separated by two-dimensional polyacrylamide gel electrophoresis (2DE), followed by statistical analysis of the stained gels. Spots of interest were excised from the gels, digested with trypsin and identified by MALDI-TOF-TOF mass spectrometry. Results Twenty-six differential spots were detected between the two classes of tissues, among which twenty were identified by mass spectrometry and sixteen were found to be non-redundant. Eleven proteins were either underexpressed or undetected in the ccRCC extracts, such as prohibitin and peroxiredoxin-3, whereas five were found to be overexpressed or exclusively detected in the ccRCC extract, including αβ crystalin and heat shock protein 27. CONCLUSIONS Several proteins were detected at differential levels when compared to normal adjacent tissues, and, moreover, many have been previously described by their relationship with RCC. Therefore, this work corroborates previous reports on the search for biomarkers for ccRCC, as well as it points out new candidates that may be validated in future studies

Proteomics analysis of tissue samples from patients with squamous cell carcinoma of the penis and positive to human papillomavirus

ABSTRACTPurpose:The aim of this study was to identify possible protein biomarkers and/or candidates for therapeutic targets in tissues of patients with SCCP, infected by HPV, applying one dimensional electrophoresis (1DE), followed by direct mass spectrometry (MS) analysis.Materials and Methods:Tissues from 10 HPV positive patients with SCCP and from 10 patients with HPV negative non-tumorous penile foreskins were analyzed applying 1D electrophoresis, followed by analysis with direct mass spectrometry (MS).Results:Sixty-three different proteins were identified in the first group and 50 in the second group. Recognition was possible for 28 proteins exclusively detected in Group 1 and 21 proteins presented only in Group 2.Conclusion:Some proteins in the first group are directly involved in the development of other types of cancer, and therefore, suitable for analysis. Complement C3 protein is a strong candidate for evaluating SCCP patients

Characterization and internalization of small extracellular vesicles released by human primary macrophages derived from circulating monocytes.

Extracellular vesicles (EVs) are small membrane-limited structures derived from outward budding of the plasma membrane or endosomal system that participate in cellular communication processes through the transport of bioactive molecules to recipient cells. To date, there are no published methodological works showing step-by-step the isolation, characterization and internalization of small EVs secreted by human primary macrophages derived from circulating monocytes (MDM-derived sEVs). Thus, here we aimed to provide an alternative protocol based on differential ultracentrifugation (dUC) to describe small EVs (sEVs) from these cells. Monocyte-derived macrophages were cultured in EV-free medium during 24, 48 or 72 h and, then, EVs were isolated from culture supernatants by (dUC). Macrophages secreted a large amount of sEVs in the first 24 h, with size ranging from 40-150 nm, peaking at 105 nm, as evaluated by nanoparticle tracking analysis and scanning electron microscopy. The markers Alix, CD63 and CD81 were detected by immunoblotting in EV samples, and the co-localization of CD63 and CD81 after sucrose density gradient ultracentrifugation (S-DGUC) indicated the presence of sEVs from late endosomal origin. Confocal fluorescence revealed that the sEVs were internalized by primary macrophages after three hours of co-culture. The methodology here applied aims to contribute for enhancing reproducibility between the limited number of available protocols for the isolation and characterization of MDM-derived sEVs, thus providing basic knowledge in the area of EV methods that can be useful for those investigators working with sEVs released by human primary macrophages derived from circulating monocytes