Diversity assessment of benthic macroinvertebrates, yeasts, and microbiological indicators along a longitudinal gradient in Serra do Cipó, Brazil.

The main goals of this study were: 1) to evaluate the structure, diversity, and functional trophic group composition of benthic macroinvertebrate communities; 2) to characterize water quality in the headwaters of the Doce river watershed, based on physical, chemical, and biological parameters (benthic macroinvertebrates, fecal coliforms, heterotrophic bacteria, and yeasts); and 3) to contribute to the knowledge of the structure and function of longitudinal gradients in lotic ecosystems in Brazil. A total of 60 benthic macroinvertebrate taxa were identified, the dominant group being the aquatic insects, with 50 families distributed in 8 orders. The dry period presented higher values of taxonomic richness and total density of benthic macroinvertebrates. A decreasing gradient was observed in these variable values from the 3rd order stretch down to the 6th order stretch. The highest Shannon-Wiener diversity values were found in the rainy period in the 3rd order stretches, which presented well-developed riparian forest. Besides the 3rd order stretches, the Pielou evenness index values were also high in the 6th order stretch. The collectors, together with the scrapers, predominated in the benthic macroinvertebrate communities in all river stretches, except in the 2nd, 4th, and 5th order stretches in the rainy period, where communities were dominated by filterers. The shredders and predators presented low densities for all river stretches. All microbiological variables presented low levels. Due to the high counts of heterotrophic bacteria and coliforms, the studied river stretches presented inadequate potability but adequate balneability levels. The results suggest that the structure, diversity, and composition of the benthic macroinvertebrate communities are influenced by the trophic resource availability, seasonality, and sediment heterogeneity. The microbiological results of this study allow inferring that the waters from Serra do Cipó have excellent potential for recreational use and as future sources of water for human consumption.     

Fructose-1,6-bisphosphate reverts iron-induced phenotype of hepatic stellate cells by chelating ferrous ions

Hepatic fibrosis is an extracellular matrix deposition by hepatic stellate cells (HSC). Fibrosis can be caused by iron, which will lead to hydroxyl radical production and cell damage. Fructose-1,6-bisphosphate (FBP) has been shown to deliver therapeutic effects in many pathological situations. In this work, we aimed to test the effects of FBP in HSC cell line, GRX, exposed to an excess of iron (Fe). The Fe-treatment increased cell proliferation and FBP reversed this effect, which was not due to increased necrosis, apoptosis or changes in cell cycle. Oil Red-O staining showed that FBP successfully increased lipid content and lead GRX cells to present characteristics of quiescent HSC. Fe-treatment decreased PPAR-γ expression and increased Col-1 expression. Both effects were reversed by FBP which also decreased TGF-β1 levels in comparison to both control and Fe groups. FBP, also, did not present scavenger activity in the DPPH assay. The treatment with FBP resulted in decreased proliferation rate, Col-1 expression and TGF-β1 release by HSC cells. Furthermore, activated PPAR-γ and increased lipid droplets induce cells to become quiescent, which is a key event to reversion of hepatic fibrosis. FBP also chelates iron showing potential to improve Cell redox state     

Lack of lactate-proton symport activity in pck1 mutants of Saccharomyces cerevisiae

Abstract Mutants of Saccharomyces cerevisiae without phosphoenolpyruvate carboxykinase activity showed no measurable lactate proton symport, while mutants without fructose-1,6-bisphosphatase had normal transport activity. Incubation of a pck1 mutant, under derepression conditions in the presence of glycerol, restored the activity of the lactate-proton symport, with identical kinetic characteristics to that in the wild-type. For efficient lactate-proton symport activity, not only is an external inducer such as lactic acid needed, but also a molecule derived from the acid metabolism may be necessary.     

Cpt1a silencing in AgRP neurons improves cognitive and physical capacity and promotes healthy aging in male mice

Orexigenic neurons expressing agouti-related protein (AgRP) and neuropeptide Y in the arcuate nucleus (ARC) of the hypothalamus are activated in response to dynamic variations in the metabolic state, including exercise. We previously observed that carnitine palmitoyltransferase 1a (CPT1A), a rate-limiting enzyme of mitochondrial fatty acid oxidation, is a key factor in AgRP neurons, modulating whole-body energy balance and fluid homeostasis. However, the effect of CPT1A in AgRP neurons in aged mice and during exercise has not been explored yet. We have evaluated the physical and cognitive capacity of adult and aged mutant male mice lacking Cpt1a in AgRP neurons (Cpt1a KO). Adult Cpt1a KO male mice exhibited enhanced endurance performance, motor coordination, locomotion, and exploration compared with control mice. No changes were observed in anxiety-related behavior, cognition, and muscle strength. Adult Cpt1a KO mice showed a reduction in gastrocnemius and tibialis anterior muscle mass. The cross-sectional area (CSA) of these muscles were smaller than those of control mice displaying a myofiber remodeling from type II to type I fibers. In aged mice, changes in myofiber remodeling were maintained in Cpt1a KO mice, avoiding loss of physical capacity during aging progression. Additionally, aged Cpt1a KO mice revealed better cognitive skills, reduced inflammation, and oxidative stress in the hypothalamus and hippocampus. In conclusion, CPT1A in AgRP neurons appears to modulate health and protects against aging. Future studies are required to clarify whether CPT1A is a potential antiaging candidate for treating diseases affecting memory and physical activity.     

A simple and economical modification of the Masson-Fontana method for staining melanin granules and enterochromaffin cells

Enterochromaffin cells from the small intestine of man, guinea pig, dog, chicken, rabbit, cat and rat were stained using the Masson-Fontana ammoniacal silver method with varying dilutions of silver nitrate solution (0.25 to 5 g per 100 ml of distilled water) and incubation temperatures (60 C and 75 C). The 0.5% solution of silver nitrate gave an argentaffin pattern similar to that of the 5% solution and had two major advantages: economically, since much less silver nitrate is used, and methodologically, since low background resulted with tissue of those species (rat, cat and rabbit) that required unusually long incubation. The staining of melanocytes was similar for all dilutions at the usual staining time (15-30 min).     

A Simple and Economical Modification of the Masson-Fontana Method for Staining Melanin Granules and Enterochromaffin Cells

Enterochromaffin cells from the small intestine of man, guinea pig, dog, chicken, rabbit, cat and rat were stained using the Masson-Fontana ammoniacal silver method with varying dilutions of silver nitrate solution (0.25 to 5 g per 100 ml of distilled water) and incubation temperatures (60 C and 75 C). The 0.5% solution of silver nitrate gave an argentaffin pattern similar to that of the 5% solution and had two major advantages: economically, since much less silver nitrate is used, and methodologically, since low background resulted with tissue of those species (rat, cat and rabbit) that required unusually long incubation. The staining of melanocytes was similar for all dilutions at the usual staining time (15-30 min).     

Description of different stages of oogenesis in Ophidion barbatum (Pisces,Ophidiidae)

Ophidion barbatum is an oviparous fish species, with external fertilization. In females, a single-lobed ovary is followed by a short oviduct that opens directly out through the genital opening. Ovarian development is asynchronous. In a mature ovary and during the breeding season, oocytes in various stages of development can be observed simultaneously. Six stages are described in the oogenesis from the oogonia to the mature egg, using different histological techniques and based on differences of staining, of size and on the nucleus and cytoplasm structure, as viewed through a light microscope. Three of them correspond to the previtellogenic phase and the other three to the vitellogenic phase. Atretic degeneration of eggs is also described.     

Dissociation kinetics of an enzyme-inhibitor system using single-molecule force measurements

We report on an improved method to interpret single molecule dissociation measurements using atomic force microscopy. We describe an easy to use methodology to reject nonspecific binding events, as well as estimating the number of multiple binding events. The method takes nonlinearities in the force profiles into account that result from the deformation of the used polymeric linkers. This new method is applied to a relevant enzyme-inhibitor system, latent matrix metalloprotease 9 (ProMMP-9, a gelatinase), and its inhibitor, tissue inhibitor of metalloproteases 1 (TIMP 1), which are important players in cancer metastasis. Our method provides a measured kinetic off-rate of 0.010 ± 0.003 s(-1) for the dissociation of ProMMP9 and TIMP1, which is consistent with values measured by ensemble methods.     

A panel of microsatellite markers for genetic studies of European polecats (Mustela putorius) and ferrets (Mustela furo)

We report a panel of 12 microsatellite markers, including nine novel polymorphic loci isolated in the European polecat and three loci previously developed in closely related species, for genetic studies of polecats and ferrets. We tested the panel at fine geographic scales in polecat and domestic ferret populations of Britain and Portugal and at a broad geographic scale, using 50 polecat samples from across Western Europe. At the fine geographic scales, we recorded one to five alleles per locus and observed and expected heterozygosities ranging from 0 to 0.80 and from 0 to 0.74, respectively. These values increased markedly in the European sample of polecats. No linkage disequilibrium was detected between any pair of loci and all genotypic frequencies complied with Hardy–Weinberg equilibrium expectations. The microsatellite panel allowed the molecular discrimination of polecats and domestic ferrets in Britain, where hybridisation between the two species has been documented. Thus, these loci may be useful not only in genetic studies of polecats and ferrets but also in studies to assess hybridisation between these two mustelid species.