NHERF1/EBP50 immunoexpression in renal cell carcinomas and oncocytomas with ultrastructural analysis of clear cell renal cell carcinoma.

Background: Na+/H+ exchanger (NHE) maintains the alkaline pH of epithelial cells working at the cellular membrane and exchanging H+/Na+ ions. In renal tubular epithelial cells, the reabsorption of NaCl is implemented by NHE3 isoform, which is regulated by NHE regulatory factor-1 (NHERF1). Normally situated at the apical zones of proximal tubular cells, NHERF1 participates in cytoskeletal reorganization and signal transduction facilitating structural stability and ion exchange. Based on an extensive search in English literature, NHERF1/EBP50 immunoexpression has been studied in breast, colon, and other tumors with only one study on 21 cases of renal cell carcinomas (RCC). Methods: Using NHERF1/EBP50 immunohistochemistry (IHC) on 64 (82%) RCCs (34 clear cells, 21 papillary and 9 chromophobe types) and 14 (18%) oncocytomas, we evaluated and scored NHERF1/EBP50 immunoexpression depending on the World Health Organization (WHO)/International Society of Urological Pathology (ISUP) grading system followed by ultrastructural identification of microlumen-like structures (MLS) in clear cell renal cell carcinomas (ccRCC). Results: Staining patterns varied throughout the tumors and within individual tumors. Only ccRCC showed unique MLS within the cytoplasm of tumor cells. All neoplasia-transformed tubular cells, regardless of the tumor grade and stage, had altered immunoexpression of NHERF1/EBP50 ranging from complete absence to aberrant expression in the luminal cell membrane, nuclear or cytoplasmic localizations. Conclusions: Only ccRCC showed unique dot-like condensations of immunostaining/MLS at membranous, submembranous, and paranuclear localizations. The latter two localizations were mainly observed in the combined WHO/ISUP grade 1 and 2 group compared to the combined group of grade 3 and 4 tumor samples (P=0.0146 and P<0.0001, respectively). Ultrastructurally, the MLS were identified as thick microvilli trapped by a single-layer membrane, displaced into the cytoplasm and ranging from 400 nm to 3.5 µm. These significant ultrastructural reorganizations may contribute to tumor progression, metastasis, and drug resistance.

Genome-Wide Association Study Reveals Novel Candidate Genes Associated with Productivity and Disease Resistance to Moniliophthora spp. in Cacao (Theobroma cacao L.).

Cacao (Theobroma cacao L.), the source of chocolate, is one of the most important commodity products worldwide that helps improve the economic livelihood of farmers. Diseases like frosty pod rot caused by Moniliophthora roreri and witches' broom caused by Moniliophthora perniciosa limit the cacao productivity, this can be solved by using resistant varieties. In the current study, we sequenced 229 cacao accessions using genotyping-by-sequencing to examine the genetic diversity and population structure employing 9,003 and 8,131 single nucleotide polymorphisms recovered by mapping against two cacao genomes (Criollo B97-61/B2 v2 and Matina 1-6 v1.1). In the phenotypic evaluation, three promising accessions for productivity and 10 with good tolerance to the frosty pod rot and witches' broom diseases were found. A genome-wide association study was performed on 102 accessions, discovering two genes associated with productivity and seven to disease resistance. The results enriched the knowledge of the genetic regions associated with important cacao traits that can have significant implications for conservation and breeding strategies like marker-assisted selection.

Proteomic and peptidomic profiling of Brazilian artisanal 'Coalho' cheese.

BACKGROUND: Artisanal 'Coalho' cheese is a product typically popular in the Brazilian north-eastern region. Production of this cheese represents about 9.2% of the internal crude product of Pernambuco State. Several peptides are generated from hydrolysis of αS1 -, αS2 -, ß-, and κ-caseins during manufacture of this cheese. The commercial importance of Brazilian artisanal 'Coalho' cheese justifies the examination of both the protein and peptide profiles of cheeses from six cities of the semi-arid region of Pernambuco State, Brazil. RESULTS: SDS-PAGE of the aqueous extracts of 'Coalho' cheeses (WSP) showed bands of lactoferrin, ß-lactoglobulin, ß-lactoglobulin (dimer), α-lactoalbumin, bovine serum albumin, α-casein, ß-casein, κ-casein and para-κ-casein. A total of 57 to 72 peptides were confirmed by mass spectra in the different samples of 'Coalho' cheese which 32 known peptides (11 from αS1 -casein, three from αS2 -casein, 15 from ß-casein and three from κ-casein), comprising seven caseinphosphopeptides. Among the unidentified peptides, three showed high intensity peaks in all 'Coalho' cheeses studied (with molecular weights of 1597, 1725/1726, 2778/2779 Da). CONCLUSION: The proteomic studies revealed peptides that may represent molecular markers or fingerprints for investigating the quality control and regional characterisation of these 'Coalho' cheeses. © 2016 Society of Chemical Industry.

Intratracheal co-administration of antioxidants and ceftriaxone reduces pulmonary injury and mortality rate in an experimental model of sepsis.

BACKGROUND AND OBJECTIVE: Recent studies showed that both sepsis and antibiotic therapy are associated with cell death and linked to reactive oxygen species generation. This study investigated the effects of intratracheal administration of combinations of antioxidants (n-acetyl cysteine (NAC), vitamins C and E) in the treatment of sepsis-induced lung injury. METHODS: Ninety-six male Wistar rats subjected to sepsis were treated with ceftriaxone plus NAC with or without vitamins C and E and compared to appropriate controls. As an index of oxidative damage protein carbonyls, sulfhydryl groups, lipid peroxidation and superoxide anion were measured, as well as superoxide dismutase and catalase. Histopathological alterations and mortality rate were also analyzed. RESULTS: Twenty-four hours after sepsis induction, markers of oxidative stress increased in all lungs examined. Ceftriaxone plus intratracheal combination of NAC, vitamins C and E decreased lung injury in infected animals by reducing superoxide anion production (54%), lipid peroxidation (53%) and protein carbonyl (58%) and restored the redox status (7.5 times). This therapy also reduced the imbalance of antioxidant enzymes activities and attenuated the alveolar architectural disorganization, inflammatory cell infiltration and pulmonary oedema. Survival increased from 66.6% with ceftriaxone to 83.2% with ceftriaxone plus antioxidants. CONCLUSIONS: Ceftriaxone plus intratracheal co-administration of antioxidants provides better protection, by decreasing pulmonary oxidative stress, limiting histophatological alterations and improving survival. Antioxidants should be explored as a co-adjuvant in the treatment of severe lung injury.

Kinetics and bioreactor studies of immobilized invertase on polyurethane rigid adhesive foam.

A new support, polyurethane rigid adhesive foam (PRAF), which can be used to cover internal surface of metallic tubes, was used to immobilize invertase for application in an enzymatic bioreactor. The kinetic parameters were: Km--46.5±1.9 mM (PRAF-invertase) and 61.2±0.1 mM (free enzyme) and Vmax 42.0±4.3 U/mg protein/min (PRAF-invertase) and 445.3±24.0 U/mg protein/min (free invertase). The PRAF-invertase derivative maintained 50.1% of initial activity (69.17 U/g support) for 8 months (4°C) and was not observed microbial contamination. The bioreactor showed the best production of inverted sugar syrup using up-flow rate (0.48 L/h) with average conversion of 10.64±1.5% h(-1) at feeding rate (D) of 104 h(-1). The operational inactivation rate constant (kopi) and half-life were 1.92×10(-4) min(-1) and 60 h (continue use). The PRAF spray support looks promising as a new alternative to produce immobilized derivatives on reactor surfaces.

Bovine trypanosomosis due to trypanosoma vivax in the German Bush province Bolivia

Trypanosoma vivax es un hemoparasito encontrado en la región de la mosca tsé-tsé en Africa. Sin embargo, el se ha difundido a otras partes de Africa, Centro-América, Sud-América, Indias Occidentales e Islas Mauricio. Este trabajo es un relato de la primera ocurrencia de T. vivax en la provincia de German Bush, Bolivia. T. vivax fue identificado en 45 por ciento de los 80 bovinos examinados por el test de microhematocrito. Los síntomas clínicos observados fueran fiebre, anemia, abortos, emagrecimiento progresivo, pérdida substancial de peso en tiempo corto y emaciación progresiva y linfonodos aumentados. Los resultados de este estudio sugieren que la difusión acelerada de T. vivax podrían representar un serio impacto a la economía de la región