Beta-globin gene cluster haplotypes in two North American indigenous populations

Haplotypes derived from five polymorphic restriction sites were determined in 50 Carrier-Sekani and 70 Mvskoke chromosomes, and the results were integrated with those previously obtained for 11 South American Indian populations. Eleven haplotypes were identified in the Mvskokes, while five were observed in the Carrier-Sekani. As in South American natives, haplotype 2 (+----) and 6 (-++ -+) were the most prevalent among the Mvskoke (46% and 30%, respectively). In the Carrier-Sekani, haplotype 2 was also the most common, but haplotype 5 (-+ -++) was somewhat more frequent (18%) than 6 (12%). High heterozygosities, as well as genetic differentiation, were observed among these two North American and two other South American groups (Mapuche and Xavante). They could be due to non-Indian admixture in the Mvskoke and Mapuche, but the findings in the other two populations require some other type of explanation.     

Harvesting for food versus feed: a review of Peruvian fisheries in a global context

Peru is the top exporter of fishmeal and fish oil (FMFO) worldwide and is responsible for half and a third of global production, respectively. Landings of “anchoveta” (Engraulis ringens) are used nearly exclusively for FMFO production, despite a proactive national food policy aimed at favoring the direct human consumption of this inexpensive species. It may be surprising that in a country where malnutrition and caloric deficit constitute major issues, a low-priced and highly nutritious fish such as anchovy does not have stronger domestic demand as a food fish. Here, we review and assess eight potential politico-socio-economic processes that can explain this situation. The main explanation are dietary habits, the preference for broiler and the higher profit from anchovy sold as feed fish compared to its use as a food fish due to historically high FMFO prices, boosted by an increasing demand for aquaculture in a context of finite forage and trash fish resources. In addition, the recent introduction of an individual quota system has shifted bargaining power from processors to fishers, thereby increasing competition for the raw material. This competition results in an increase in anchovy prices offered by the feed fish industry due to its onshore processing overcapacity, which is detrimental to the food fish industry. In the end, although the dominant use of anchovy for fish feed is largely explained by integrating these market mechanisms and other minor ones, this use raises other issues, such as rent redistribution through public policies, employment, equitability and utility (low social costs), and resource management (threats to ecosystems or global change). Different policy scenarios are proposed in relation to these issues.     

The synergistic risk effect of apolipoprotein ε4 and DNA (cytosine-5-)-methyltransferase 3 beta (<Emphasis Type="Italic">DNMT3B</Emphasis>) haplotype for Alzheimer’s disease

Alzheimer’s disease (AD) is a complex and multifactorial disease with the contribution of several genes and polymorphisms to its development. Among these genes, the APOEε4 is the best known risk factor for AD. Methylation is associated with APOE expression and AD development. Recently, we found an association of the TGG haplotype in the DNMT3B gene, one of the catalyst enzyme for methylation, with AD. Therefore, the objective of the study was to investigate whether APOEε4 and TGG haplotype have an synergistic effect on AD. The sample was composed of 212 Caucasian individuals (108 healthy controls and 104 with AD by NINCDS-ADRDA and DSM-IV-TR criteria) from southern Brazil. The genetic analyses were performed by real time PCR for TaqMan^® assay. Multivariate logistic regression was performed categorizing groups according to presence of APOEε4 and/or TGG haplotype as an independent variable for outcome AD. The presence of TGG haplotype plus the allele APOEε4 were strongly associated with AD [OR 11.13; 95 % CI (4.25–29.16); P < 0.001]. This association had a higher risk than each risk factor alone. We found a strong association of the interaction of DNMT3B gene with the APOEε4 in this sample of AD patients. The presence of TGG haplotype and APOEε4 significantly increased the risk of developing the disease, showing an synergistic effect.     

Alveolar Gradient of Pentane in Normal Human Breath

Previous studies have raised the question of whether pentane is a normal constituent of human breath, since its concentrations in inspired room air and expired breath are often similar. Using a highly sensitive assay for volatile organic compounds, we studied 37 normal subjects in order to determine the alveolar gradient of pentane in their breath (i.e. concentration in alveolar breath minus concentration in the inspired air). The chemical identity of pentane was confirmed by mass spectroscopy. The alveolar gradient of pentane was zero ± 0.175 nmol/1 in 54.1 % of subjects, and distributed in an approximately bell-shaped curve. Determination of the alveolar gradient divided the normal subjects into three groups: the “passive equilibrators” who did not appear to excrete pentane in the breath (the majority), “metabolizers” who actively catabolized inhaled pentane, and “manufacturers” who excreted more pentane than they inhaled.     

Response to treatment in Brazilian patients with chronic hepatitis C is associated with a single-nucleotide polymorphism near the interleukin-28B gene

A single-nucleotide polymorphism (SNP) upstream of interleukin (IL)28B was recently identified as an important predictor of the outcome of chronic hepatitis C patients treated with pegylated interferon plus ribavirin (PEG-IFN/RBV). The aim of this study was to investigate the association between the IL28B gene polymorphism (rs12979860) and virological response in chronic hepatitis C patients. Brazilian patients (n = 263) who were infected with hepatitis C virus (HCV) genotype 1 and were receiving PEG-IFN/RBV were genotyped. Early virological response (EVR) (12 weeks), end-of-treatment response (EOTR) (48 weeks), sustained virological response (SVR) (72 weeks) and relapse were evaluated using conventional and quantitative polymerase chain reaction (PCR) assays. The frequency of the C allele in the population was 39%. Overall, 43% of patients experienced SVR. The IL28B CC genotype was significantly associated with higher treatment response rates and a lower relapse rate compared to the other genotypes [84% vs. 58% EVR, 92% vs. 63% EOTR, 76% vs. 38% SVR and 17% vs. 40% relapse rate in CC vs. other genotypes (CT and TT), respectively]. Thus, the IL28B genotype appears to be a strong predictor of SVR following PEG-IFN/RBV therapy in treatment-naïve Brazilian patients infected with HCV genotype 1. This study, together with similar research examining other SNPs, should help to define adequate protocols for the treatment of patients infected with HCV genotype 1, especially those with a poor prognosis.     

PPARA, RXRA, NR1I2 and NR1I3 gene polymorphisms and lipid and lipoprotein levels in a Southern Brazilian population

Cardiovascular disease is the main cause of death worldwide, and dyslipidemia is an important multifactorial risk factor. Considering the involvement of nuclear receptors in metabolic pathways, and that some of the receptors act in lipid metabolism and homeostasis, the aim of the present study was to investigate the influence of genetic variations in RXRA, PPARA, NR1I2, and NR1I3 on lipid and lipoprotein levels. Five polymorphisms in the aforementioned genes were genotyped in 622 Brazilians of European descent by PCR-RFLP or TaqMan genotyping assays. In general, carriers of the A insertion of RXRA rs11381416 polymorphism showed higher levels of triglyceride (TG; 1.80 ± 1.20 vs. 1.52 ± 1.20 mmol/L; P = 0.020). Moreover, sexual dimorphic association was found (gender*NR1I3 rs2501873 genotype interaction P < 0.001), males with NR1I3 rs2501873 G/G genotype had lower TG levels (ANCOVA, P = 0.009). Our results suggest that polymorphisms in the RXRA and NR1I3 genes influence lipid profile in a Southern Brazilian population. However, these general and gender association require confirmation in subsequent studies.     

In vitro culture supplementation of EGF for improving the survival of equine preantral follicles

Folliculogenesis is a process of development and maturation of the ovarian follicles, being essential for the maintenance of fertility. In in vivo conditions, 99.9% of the follicles of an ovary do not ovulate and undergo atresia. In order to minimize this loss and to clarify the existing mechanisms, a technique was developed that allows for the in vitro follicular development. The objective of this study was to evaluate the effects of different epidermal growth factor (EGF) concentrations on the in vitro culturing of equine preantral follicles. Ovaries (n?=?10) were collected from a local slaughterhouse of mares in seasonal anestrus, washed with 70% alcohol and PBS, and transported. The inner portion of the ovary was divided into 11 fragments of approximately 3?×?3?×?1 mm. A fragment of each ovary was immediately fixed in Bouin (control group). The remaining 10 fragments were individually cultured for 2 and 6 d. The medium was supplemented with different concentrations of EGF (0, 10, 50, 100, and 200 ng/mL). After cultivation, the fragments were processed and classified according to the developmental stage and morphology. In total, 1065 slides containing 6105 tissue sections were evaluated. Within 2 d of culture, there was a higher proportion of intact follicles at the EGF concentrations of 0 and 100 ng/mL (p?>?0.05). After 6 d of culture, only the EGF concentration of 100 ng/mL demonstrated a difference when compared to the other treatments (0, 10, 50 and 200 ng/mL of EGF, p?>?0.05). There was follicular development after 2 d at all EGF concentrations. Thus, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in in vitro cultures of ovarian fragments for 2 d. In addition, we suggest that EGF promotes follicular survival in equines at a concentration of 100 ng/mL in situ cultivation.     

The Macrophage Activation Induced by Bacillus thuringiensis Cry1Ac Protoxin Involves ERK1/2 and p38 Pathways and the Interaction with Cell‐Surface‐HSP70

Here, we aimed to further characterize the mechanisms involved in protoxin (p) Cry1Ac‐induced macrophage activation. We demonstrated that pCry1Ac induces MAPK ERK1/2, p38, and JNK phosphorylation in RAW264.7 macrophages. Because MAPK activation is mainly triggered via ligand–receptor interactions, we focused on the identification of potential pCry1Ac‐receptor proteins. Flow cytometry and confocal analysis showed specific saturable pCry1Ac‐binding to the macrophage surface and evidenced its internalization via the clathrin‐pathway. We performed immunoprecipitation assays and identified by MALDI‐TOF‐TOF several possible pCry1Ac‐binding proteins, such as heat shock proteins (HSPs), vimentin, α‐enolase, and actin; whose interaction and presence was confirmed, respectively, by ligand blot and Western blot assays. We also detected cell‐surface (cs) pCry1Ac‐HSP70 colocalization, so HSP70 was chosen for further characterization. Co‐immunoprecipitation with HSP70 antibodies followed by Western blot confirmed the pCry1Ac‐HSP70 interaction. Furthermore, pretreatment of RAW264.7 cells with HSP70 antibodies reduced pCry1Ac‐induced ERK1 phosphorylation and MCP‐1 production; thus suggest the functional participation of csHSP70 in pCry1Ac‐induced macrophage activation. csHSP70 also was evaluated in peritoneal‐cavity (PerC) macrophages of untreated BALB/c mice, interestingly it was found that the predominant population namely large‐peritoneal‐macrophages (LPM) displayed csHSP70 + hi. Furthermore, the dynamics of PerC macrophage subsets, LPM, and small‐peritoneal macrophages (SPM) were evaluated in response to in vivo pCry1Ac stimuli in presence or not of phenylethynesulfonamide (PES) a functional HSP70 inhibitor. It was found that pCry1Ac increased the proportion of SPM CD11b + F4/80 + lowMHCII + csHSP70 + low and markedly reduced the amount of LPM CD11b + F4/80 + hiMHCII‐csHSP70 + hi; while PES, partially suppressed this pCry1Ac‐induced effect, further suggesting the participation of HSP70 in macrophage activation process. J. Cell. Biochem. 119: 580–598, 2018. © 2017 Wiley Periodicals, Inc.     

Verification of strain identity in Brazilian soybean inoculants by using the Polymerase Chain Reaction

The standards for meeting inoculant quality in Brazil include a requirement of strain verification as well as the necessity for a minimal number of viable bacterial cells. This requirement stimulated us to develop a rapid and simple PCR-based method to distinguish the four Brazilian strains of Bradyrhizobium, SEMIA 587, SEMIA 5019 (29W), SEMIA 5079 (CPAC-15) and SEMIA 5080 (CPAC-7), recommended for the inoculation of soybean. PCR reactions using the random amplified polymorphic DNA (RAPD) PCR primer CRL-7 together with a modified buffer resulted in four different fingerprint patterns, which permitted the Brazilian soybean strains to be distinguished from each other. The fingerprint patterns obtained were reproducible irrespective of the source of templates. Comparisons with fingerprint patterns obtained with soybean bradyrhizobia from the USDA Culture Collection led to the conclusion that two of the Brazilian inoculant strains (SEMIA 587 and SEMIA 5019) were characteristic for B. elkanii, while the other two (SEMIA 5079 and SEMIA 5080) were more typical of B. japonicum. Also, we obtained evidence that the RAPD analysis with primer CRL-7 may be useful for distinguishing variants of the same strain.