Inhibition of the LOX enzyme family members with old and new ligands. Selectivity analysis revisited

Lysyl oxidase (LOX) enzymes as potential drug targets maintain constant attention in the therapy of fibrosis, cancer and metastasis. In order to measure the inhibitory activity of small molecules on the LOX enzyme family members a fluorometric activity screening method was developed. During assay validation, previously reported non-selective small inhibitor molecules (BAPN, MCP-1, thiram, disulfiram) were investigated on all of the major LOX enzymes. We confirmed that MCP-1, thiram, disulfiram are in fact pan-inhibitors, while BAPN inhibits only LOX-like enzymes (preferably LOX-like-protein-2, LOXL2) in contrast to the previous reports. We measured the LOX inhibitory profile of a small targeted library generated by 2D ligand-based chemoinformatics methods. Ten hits (10.4% hit rate) were identified, and the compounds showed distinct activity profiles. Potential inhibitors were also identified for LOX-like-protein-3 (LOXL3) and LOX-like-protein-4 (LOXL4), that are considered as emerging drug targets in the therapy of melanoma and gastric cancer.     

Elucidating the role of oxidative stress in the therapeutic effect of rutin on experimental acute pancreatitis

Introduction: Acute pancreatitis (AP) may be severe and cause hospitalization or death, and the available treatment is insufficient to control pancreatic inflammation and pain. Rutin is a natural flavonoid with the potential to treat AP via anti-inflammatory, antinociceptive, and antioxidant activities.

Aim: This study investigated the beneficial effects of rutin on experimental AP induced by l-arginine administration in mice.

Methods: The l-arginine-induced AP model was used in Swiss mice (n?=?6–8). Mice submitted to AP induction were treated with rutin (37.5, 75, or 150?mg kg^?1, p.o.) or vehicle (saline) after 24, 36, 48, and 60?h of AP induction. Abdominal hyperalgesia, serum enzymes, interleukin (IL)-6 levels, pancreatic inflammatory parameters, malondialdehyde (MDA) levels, antioxidant enzyme activities, and 3-nitrotyrosine contents were measured 72?h after induction.

Results: Mice submitted to l-arginine injections developed abdominal hyperalgesia and increased serum amylase, lipase, C-reactive protein and IL-6 concentrations; and increased pancreatic myeloperoxidase activity, edema index, MDA, and 3-nitrotyrosine contents. A marked decrease in catalase activity was observed in the pancreas without alterations of superoxide dismutase (SOD) activity compared with the control group. Rutin treatment significantly impaired all the parameters that were altered by AP induction, but increased catalase and SOD activities in the pancreas compared with the vehicle-treated group.

Conclusion: Rutin treatment exerted a protective effect on l-arginine-induced AP by mechanisms involving the reduction of oxidative stress, which suggests that this flavonoid has a potential for future approaches designed for the management of AP.     

Guarana (Paullinia cupana var. sorbilis), an anciently consumed stimulant from the Amazon rain forest: the seeded-fruit transcriptome

Guarana (Paullinia cupana var. sorbilis) is a plant native to the central Amazon basin. Roasted seed extracts have been used as medicinal beverages since pre-Colombian times, due to their reputation as stimulants, aphrodisiacs, tonics, as well as protectors of the gastrointestinal tract. Guarana plants are commercially cultivated exclusively in Brazil to supply the national carbonated soft-drink industry and natural product stores around the world. In this report, we describe and discuss the annotation of 15,387 ESTs from guarana seeded-fruits, highlighting sequences from the flavonoid and purine alkaloid pathways, and those related to biotic stress avoidance. This is the largest set of sequences registered for the Sapindaceae family.     

Characterization of a novel Obg-like ATPase in the protozoan Trypanosoma cruzi

We characterized a gene encoding an YchF-related protein, TcYchF, potentially associated with the protein translation machinery of Trypanosoma cruzi. YchF belongs to the translation factor-related (TRAFAC) class of P-loop NTPases. The coding region of the gene is 1185 bp long and encodes a 44.3 kDa protein. BlastX searches showed TcYchF to be very similar (45-86%) to putative GTP-binding proteins from eukaryotes, including some species of trypanosomatids (Leishmania major and Trypanosoma brucei). A lower but significant level of similarity (38-43%) was also found between the predicted sequences of TcYchF and bacterial YyaF/YchF GTPases of the Spo0B-associated GTP-binding protein (Obg) family. Some of the most important features of the G domain of this family of GTPases are conserved in TcYchF. However, we found that TcYchF preferentially hydrolyzed ATP rather than GTP. The function of YyaF/YchF is unknown, but other members of the Obg family are known to be associated with ribosomal subunits. Immunoblots of the polysome fraction from sucrose gradients showed that TcYchF was associated with ribosomal subunits and polysomes. Immunoprecipitation assays showed that TcYchF was also associated with the proteasome of T. cruzi. Furthermore, inactivation of the T. brucei homolog of TcYchF by RNA interference inhibited the growth of procyclic forms of the parasite. These data suggest that this protein plays an important role in the translation machinery of trypanosomes.     

Expanding the Knowledge of the Geographic Distribution of Trypanosoma cruzi TcII and TcV/TcVI Genotypes in the Brazilian Amazon

Trypanosoma cruzi infection is a complex sylvatic enzooty involving a wide range of animal species. Six discrete typing units (DTUs) of T. cruzi, named TcI to TcVI, are currently recognized. One unanswered question concerning the epidemiology of T. cruzi is the distribution pattern of TcII and hybrid DTUs in nature, including their virtual absence in the Brazilian Amazon, the current endemic area of Chagas disease in Brazil. Herein, we characterized biological samples that were collected in previous epizootiological studies carried out in the Amazon Basin in Brazil. We performed T. cruzi genotyping using four polymorphic genes to identify T. cruzi DTUs: mini-exon, 1f8, histone 3 and gp72. This analysis was conducted in the following biological samples: (i) two T. cruzi isolates obtained by culturing of stools from the triatomine species Rhodnius picttipes and (ii) five serum samples from dogs in which trypomastigotes were observed during fresh blood examination. We report for the first time the presence of TcII and hybrid DTUs (TcV/TcVI) in the Amazon region in mixed infections with TcI. Furthermore, sequencing of the constitutive gene, gp72, demonstrated diversity in TcII even within the same forest fragment. These data show that TcII is distributed in the five main Brazilian biomes and is likely more prevalent than currently described. It is very probable that there is no biological or ecological barrier to the transmission and establishment of any DTU in any biome in Brazil.     

Early ontogeny of the commercially valuable fish red-bellied pacu Piaractus brachypomus (Characiformes,Serrasalmidae) from the Amazon,Brazil

The initial development of the red-bellied pacu Piaractus brachypomus is described using morphological, meristic and morphometric characteristics. A total of 127 individuals were analysed (47 in the yolk-sac, 35 in pre-flexion, five in flexion, 20 in post-flexion and 20 in juvenile) with standard length varying between 2.92 and 48.61 mm. The larvae are born poorly developed and have a discoidal yolk at ~6.33 mm standard length. During early ontogeny, the mouth passes from terminal to subterminal and the anal opening reaches the vertical line over the midline region of the body. There are changes in body shape from long and moderate to deep, head length from small to large, and eye diameter from moderate to large. Dendritic chromatophores were present in the ventral, dorsal and upper part of the swim bladder in the early larval stages. Rounded spots are evident all over the body in juveniles. The total number of myomeres ranges from 39 to 41 (20–23 pre-anal, 17–20 post-anal). Through the morphometric relationships, it was evidenced that the greatest changes during the initial ontogeny of P. brachypomus occur in the transition from the post-flexion stage to the juvenile period, indicating changes in behaviour, foraging and physiology.     

Factors Associated to Vaccination against Influenza among Elderly in a Large Brazilian Metropolis

Background

This study aimed to estimate coverage and identify factors associated to vaccination against influenza in the elderly population.

Methods

The study design was cross-sectional and population based. Data was collected in 2010 by the Health, Well-Being and Aging Study. Sample consisted of 1,341 community-dwelling elderly, in São Paulo, Brazil. Association between vaccination and covariates was evaluated by means of prevalence ratios estimated by Poisson regression models.

Results

Self-reported vaccination coverage was 74.2% (95% confidence interval: 71.3–76.9). Remaining physically active and having had recent interaction with health services, mainly with public units of healthcare, were the main incentives to increase vaccination coverage among the elderly; whereas lower age, living alone and absent interaction with health services were the main constraints to influenza vaccination at the community level. These covariates had already been reported to influence influenza vaccination of elders in previous years.

Conclusion

Previous knowledge already available on the main constraints to influenza vaccination has not allowed to remove them. Influenza campaigns should be strengthened to increase vaccination coverage, especially in the group more reticent to vaccination. Instructing healthcare providers to recommend vaccine uptake is an important piece of this puzzle.     

Early development of the Neotropical fish known as long sardine Triportheus auritus (Valenciennes 1850) (Characiformes,Triportheidae)

Larvae and juveniles of long sardine, Triportheus auritus, from the lower Amazon river was described, evaluating ontogenetic changes in their external mor­phology, pigmentation, fin development, morphometry, and meristics. A total of 93 individuals, 83 larvae and 10 juveniles were analyzed, they were captured monthly between 2014 and 2019 in the Amazon river channel and in macrophytes aquatic stands in the alluvial plains located in the lower Amazon River. From each specimen, morphometric and meristic data were measured and then the growth pattern between morphometric variables was analyzed. The larvae have an elongated body in a fusiform shape, superior mouth, simple nostril, pigmented spherical eyes and long intestine, surpassing the median region of the body. Initial pigmentation is scarce, but intensifies through development forming a pattern composed of three longitudinal bands concentrated in the ventral, cephalo-dorsal and lateral line regions. There are also pigments in the mandible, surrounding the mouth, under the swim bladder, intestine and fins. The sequence of complete fin formation is: caudal, anal, dorsal, pectoral and pelvic. The total number of myomeres ranged from 45 to 48 (25–29 preanal and 17–22 postanal). Morphometric relationships indicated differential growth for measurable morphometric parameters, with abrupt growth of snout length, head length and body height in the transition from flexion to postflexion stages. The pre-dorsal distance showed a decrease in the growth rate at the threshold from the larval to the juvenile period. The pre-pectoral and pre-anal distances showed negative allometric growth. In conclusion, the combination of body shape pigmentation pattern and, the formation sequence of fins allow the identification of the genus and coupled with the number of myomeres, morphometric relationships, and ray numbers of the anal fin ensure the differentiation of T. auritus from the other congeneric species. The metamorphosis occurred mainly at the end of the larval period and it is related to changes in the physiological and ecomorphological characteristics of the species.     

Gluten-free diet reduces adiposity,inflammation and insulin resistance associated with the induction of PPAR-alpha and PPAR-gamma expression

Gluten exclusion (protein complex present in many cereals) has been proposed as an option for the prevention of diseases other than coeliac disease. However, the effects of gluten-free diets on obesity and its mechanisms of action have not been studied. Thus, our objective was to assess whether gluten exclusion can prevent adipose tissue expansion and its consequences. C57BL/6 mice were fed a high-fat diet containing 4.5% gluten (Control) or no gluten (GF). Body weight and adiposity gains, leukocyte rolling and adhesion, macrophage infiltration and cytokine production in adipose tissue were assessed. Blood lipid profiles, glycaemia, insulin resistance and adipokines were measured. Expression of the PPAR-α and γ, lipoprotein lipase (LPL), hormone sensitive lipase (HSL), carnitine palmitoyl acyltransferase-1 (CPT-1), insulin receptor, GLUT-4 and adipokines were assessed in epidydimal fat. Gluten-free animals showed a reduction in body weight gain and adiposity, without changes in food intake or lipid excretion. These results were associated with up-regulation of PPAR-α, LPL, HSL and CPT-1, which are related to lipolysis and fatty acid oxidation. There was an improvement in glucose homeostasis and pro-inflammatory profile-related overexpression of PPAR-γ. Moreover, intravital microscopy showed a lower number of adhered cells in the adipose tissue microvasculature. The overexpression of PPAR-γ is related to the increase of adiponectin and GLUT-4. Our data support the beneficial effects of gluten-free diets in reducing adiposity gain, inflammation and insulin resistance. The data suggests that diet gluten exclusion should be tested as a new dietary approach to prevent the development of obesity and metabolic disorders.