Protective human leucocyte antigen haplotype, HLA-DRB1*01-B*14, against chronic Chagas disease in Bolivia

Background

Chagas disease, caused by the flagellate parasite Trypanosoma cruzi affects 8–10 million people in Latin America. The mechanisms that underlie the development of complications of chronic Chagas disease, characterized primarily by pathology of the heart and digestive system, are not currently understood. To identify possible host genetic factors that may influence the clinical course of Chagas disease, Human Leucocyte Antigen (HLA) regional gene polymorphism was analyzed in patients presenting with differing clinical symptoms.

Methodology

Two hundred and twenty nine chronic Chagas disease patients in Santa Cruz, Bolivia, were examined by serological tests, electrocardiogram (ECG), and Barium enema colon X-ray. 31.4% of the examinees showed ECG alterations, 15.7% megacolon and 58.1% showed neither of them. A further 62 seropositive megacolon patients who had undergone colonectomy due to acute abdomen were recruited. We analyzed their HLA genetic polymorphisms (HLA-A, HLA-B, MICA, MICB, DRB1 and TNF-alpha promoter region) mainly through Sequence based and LABType SSO typing test using LUMINEX Technology.

Principal Findings

The frequencies of HLA-DRB1*01 and HLA-B*14:02 were significantly lower in patients suffering from megacolon as well as in those with ECG alteration and/or megacolon compared with a group of patients with indeterminate symptoms. The DRB1*0102, B*1402 and MICA*011 alleles were in strong Linkage Disequilibrium (LD), and the HLA-DRB1*01-B*14-MICA*011haplotype was associated with resistance against chronic Chagas disease.

Conclusions

This is the first report of HLA haplotype association with resistance to chronic Chagas disease.     

Methylation of the DFNA5 increases risk of lymph node metastasis in human breast cancer

The pathogenesis of breast cancer involves multiple genetic and epigenetic events. In this study, we report an epigenetic alteration of DFNA5 in human breast cancer. DFNA5 gene was silenced in breast cancer cell lines that were methylated in the DFNA5 promoter, and restored by treatment with the demethylating agent, 5-aza-dC, and gene knock-down of DFNA5 increased cellular invasiveness in vitro. The mRNA expression of DFNA5 in breast cancer tissues was down-regulated as compared to normal tissues. Moreover, the DFNA5 promoter was found to be methylated in primary tumor tissues with high frequency (53%, 18/34). Quantitative methylation-specific PCR of DFNA5 clearly discriminated primary breast cancer tissues from normal breast tissues (15.3%, 2/13). Moreover, methylation status of DFNA5 was correlated with lymph node metastasis in breast cancer patients. Our data implicate DFNA5 promoter methylation as a novel molecular biomarker in human breast cancer.     

Use of 2-deoxyglucose in liquid media for the selection of mutant strains of Penicillium echinulatum producing increased cellulase and β-glucosidase activities

Mutagenesis and selection were applied to a strain of Penicillium echinulatum by treating conidia with hydrogen peroxide or 1,2,7,8-diepoxyoctane and then by incubating the conidia for 48 h in broth containing microcrystalline cellulose washed in 0.5% (w/v) aqueous 2-deoxyglucose before plating them onto cellulose agar containing 1.5% (w/v) glucose from which colonies showing the fastest production of halos of cellulose hydrolysis were selected. This process resulted in the isolation of two new cellulase-secreting P. echinulatum mutants: strain 9A02S1 showing increased cellulase secretion (2 IU ml⁻¹, measured as filter paper activity) in submerged culture in agitated flasks containing a mineral salts medium and 1% of cellulose, and strain 9A02D1, which proved more suitable for the production of cellulases in semisolid bran culture where it produced 23 IU of β-glucosidase per gram of wheat bran.     

Comparative study of the inhibitory effect of antidepressants on cholinesterase activity in Bungarus sindanus (krait) venom, human serum and rat striatum

Cholinesterases are divided into two classes based on differences in their substrate specificity and tissue distribution: acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). These enzymes may be inhibited by several compounds, such as antidepressants. The antidepressants paroxetine, imipramine, clomipramine and sertraline inhibited both venom AChE as well as human serum BChE in a concentration-dependent manner but had no effect on AChE in the rat brain striatum. The IC(50) of venom calculated for imipramine was 0.3 mM, paroxetine 0.38 mM, clomipramine 0.34 mM and sertraline 0.35 mM. Analysis of kinetic data indicated that the inhibition caused by sertraline and paroxetine was mixed, i.e. K(m) values increased and V(max) decreased in a concentration dependent manner. Imipramine and clomipramine exhibited competitive inhibition, i.e. K(m) values increased and V(max) remained constant. The present results suggest that these therapeutic agents used for depression can also be considered as inhibitors of snake venom and human serum cholinesterase.     

Macromolecular geometries determined with field-flow fractionation and their impact on the overlap concentration

In this paper we aim to understand the size/conformation relationship in waxy barley starch, a polydisperse and ultrahigh molar mass biomacromolecule. Characterizations are performed with asymmetrical flow field-flow fractionation (AsFlFFF). Furthermore, we study the effect of homogenization on the molar mass, rms radius (r rms) and hydrodynamic radius (r h). For the untreated sample, the macromolecules are elongated objects with low apparent density. As a result of homogenization, molar mass, and r rms decrease, while r h remains unaffected. The process also induces an increase, and scaling with size, of apparent density as well as changes in conformation, represented qualitatively by r rms/ r h. Finally, results from AsFlFFF are compared with viscosimetry and discussed in terms of concentration and close-packing in relation to macromolecular shape and conformation. Hence, the results show that AsFlFFF and our novel methodology enable the determination of several physical properties with high relevance for the solution behavior of polydisperse macromolecules.     

Effect of chitin waste-based composts produced by two-phase composting on two oomycete plant pathogens

Biphasic composts were prepared by first mixing peat moss and sawdust with a nitrogen-rich biomass such as chitinous waste or cow manure and composting them until termination of the thermophilic phase. These partially stabilized composts were then amended with shrimp waste inducing a second thermophilic phase. Filter-sterilized water extracts obtained from two mature biphasic composts (SP₂W₂+S and MPW+S) reduced the growth of two oomycete plant pathogens, Phytophthora fragariae var. rubi and Pythium ultimum. Both SP₂W₂+S and MPW+S composts significantly reduce the incidence of cucumber damping-off caused by Pythium ultimum as compared to a commercial brand of compost made from shrimp waste and peat moss. Hydrolysis products of chitin were unlikely to be responsible for growth inhibition since no oligomeric forms of chitin were detected in SP₂W₂+S. The shrimp waste amendment carried out after the first thermophilic phase modified the microbial populations of biphasic composts. Following the amendment, the proportion of branched-chain microbial fatty acids typical of Gram-positive bacteria increased considerably suggesting that this group of bacteria became more prevalent within the total microbial population. These data suggests that the two-phase composting process promotes the proliferation of Gram-positive bacteria antagonistic to oomycete plant pathogens.     

Ant diversity decreases during the dry season: A meta-analysis of the effects of seasonality on ant richness and abundance

Tropical studies traditionally describe insect diversity variation throughout the year. The temporally structured responses of insect assemblages to climate seasonality vary across ecosystems due to gradients of resource availability and limiting ecological factors. These idiosyncratic responses might be particularly true across the vast geographical range of the Brazilian territory, including various environments that harbor one of the most diverse ant faunas worldwide. This study addressed the relationship between ant diversity and climatic seasonality, performing a quantitative review of the published data on ant diversity collected in Brazil. We investigated the seasonality effect on ant abundance and richness described in the literature in 47 papers published between 2000 and 2018. These studies were developed mainly in the Atlantic Forest biome and collected ants with pitfall traps on the soil/litter stratum. We initially carried out a vote-counting procedure by comparing the number of significant results describing seasonal differences in the ant assemblage. We found that most papers described a similar pattern of ant abundance, richness, and species composition between seasons. However, when we performed a meta-analysis, we observed a clear pattern of higher ant abundance and richness in the wet/summer season compared with the dry/winter season. Our meta-analysis reveals that the ant diversity decreases in the dry season, strongly in the Cerrado biome. Additionally, we point out differences in the sampling effort across biomes, indicating the need for further investments in studies focused on temporal diversity patterns, including seasonal effects, on the insect assemblage in biomes less investigated so far. Abstract in Portuguese is available with online material.     

Seed- versus transplant-based eelgrass (Zostera marina L.) restoration success in a temperate marine lake

Despite active seagrass restoration gaining traction as a tool to halt and reverse worldwide seagrass losses, overall success remains limited. Restoration strategies, through seeding or transplantation, face different environmental bottlenecks that limit success. Choosing the most appropriate strategy of the two for a specific location, however, is hampered by lack of direct practical comparisons between strategies within a single system. To investigate potential life stage dependent bottlenecks, we compared seed-based and transplant-based restoration of Zostera marina in the subtidal saltwater Lake Grevelingen. Our results demonstrate that seedling recruitment was negatively impacted by bioturbation from the lugworm Arenicola marina and sediment movement due to hydrodynamic exposure. Transplant-based restoration was clearly more successful but surprisingly best predicted by leaf gluing by the ragworm Platynereis dumerilii. This previously undescribed interaction caused seagrass leaves to clump and reduce effective photosynthetic surface and leaf movement. We suggest that the observed behavior of these worms may result from a lack of foodweb interactions, illustrating the importance of  trophic control for seagrass restoration. Thus, in addition to recognizing life stage dependent environmental bottlenecks for restoration strategy selection, seagrass restoration may also require the active recovery of their associated food webs.     

Direct detection of Brucella spp. in raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes.

The 16S-23S rRNA spacer regions of Brucella abortus, B. melitensis, and B. suis were cloned and subcloned after PCR amplification. Sequence analysis of the inserts revealed a spacer of about 800 bp with very high ( > 99%) homology among the three species examined. Two genus-specific primer pairs, BRU-P5-BRU-P8 and BRU-P6-BRU-P7, that could be used in a nested PCR format and three genus-specific DNA probes, BRU-ICG2, BRU-ICG3, and BRU-ICG4, were deduced from this spacer. The specificity and sensitivity of both primer sets and probes were examined by testing them against a collection of 18 Brucella strains and 56 strains from other relevant taxa by using PCR and the Line Probe Assay (LiPA), respectively. A method for direct detection of Brucella spp. in 1 ml of raw milk was developed on the basis of enzymatic treatment of the milk components and subsequent PCR and LiPA hybridization. After a single PCR, sensitivities of 2.8 x 10(5) and 2.8 x 10(4) CFU/ml were obtained for detection by agarose gel electrophoresis and LiPA, respectively. Nested PCR yielded a sensitivity of 2.8 x 10(2) CFU/ml for both methods.