Drug Development in Conformational Diseases: A Novel Family of Chemical Chaperones that Bind and Stabilise Several Polymorphic Amyloid Structures

The increasing prevalence of conformational diseases, including Alzheimer''s disease, type 2 Diabetes Mellitus and Cancer, poses a global challenge at many different levels. It has devastating effects on the sufferers as well as a tremendous economic impact on families and the health system. In this work, we apply a cross-functional approach that combines ideas, concepts and technologies from several disciplines in order to study, in silico and in vitro, the role of a novel chemical chaperones family (NCHCHF) in processes of protein aggregation in conformational diseases. Given that Serum Albumin (SA) is the most abundant protein in the blood of mammals, and Bovine Serum Albumin (BSA) is an off-the-shelf protein available in most labs around the world, we compared the ligandability of BSA:NCHCHF with the interaction sites in the Human Islet Amyloid Polypeptide (hIAPP):NCHCHF, and in the amyloid pharmacophore fragments (Aβ17–42 and Aβ16–21):NCHCHF. We posit that the merging of this interaction sites is a meta-structure of pharmacophore which allows the development of chaperones that can prevent protein aggregation at various states from: stabilizing the native state to destabilizing oligomeric state and protofilament. Furthermore to stabilize fibrillar structures, thus decreasing the amount of toxic oligomers in solution, as is the case with the NCHCHF. The paper demonstrates how a set of NCHCHF can be used for studying and potentially treating the various physiopathological stages of a conformational disease. For instance, when dealing with an acute phase of cytotoxicity, what is needed is the recruitment of cytotoxic oligomers, thus chaperone F, which accelerates fiber formation, would be very useful; whereas in a chronic stage it is better to have chaperones A, B, C, and D, which stabilize the native and fibril structures halting self-catalysis and the creation of cytotoxic oligomers as a consequence of fiber formation. Furthermore, all the chaperones are able to protect and recondition the cerebellar granule cells (CGC) from the cytotoxicity produced by the hIAPP_20–29 fragment or by a low potassium medium, regardless of their capacity for accelerating or inhibiting in vitro formation of fibers. In vivo animal experiments are required to study the impact of chemical chaperones in cognitive and metabolic syndromes.     

Highly protective in vivo function of cytomegalovirus IE1 epitope-specific memory CD8 T cells purified by T-cell receptor-based cell sorting

Reconstitution of antiviral CD8 T cells is essential for controlling cytomegalovirus (CMV) infection after bone marrow transplantation. Accordingly, polyclonal CD8 T cells derived from BALB/c mice infected with murine CMV protect immunocompromised adoptive transfer recipients against CMV disease. The protective population comprises CD8 T cells with T-cell receptors (TCRs) specific for defined and for as-yet-unknown viral epitopes, as well as a majority of nonprotective cells with unrelated specificities. Defined epitopes include IE1/m123 and m164, which are immunodominant in terms of the magnitude of the CD8 T-cell response, and a panel of subordinate epitopes (m04, m18, M45, M83, and M84). While cytolytic T-lymphocyte lines (CTLLs) were shown to be protective regardless of the immunodominance of the respective epitope, the individual contributions of in vivo resident epitope-specific CD8 T cells to the antiviral control awaited investigation. The IE1 peptide 168-YPHFMPTNL-176 is generated from the immediate-early protein 1 (IE1) (pp89/76) of murine CMV and is presented by the major histocompatibility complex class I (MHC-I) molecule Ld. To quantitate its contribution to the protective potential of a CD8-T memory (CD8-TM) cell population, IE1-TCR+ and IE1-TCR- CD8-TM cells were purified by epitope-specific cell sorting with IE1 peptide-loaded MHC-immunoglobulin G1 dimers as ligands of cognate TCRs. Of relevance for clinical approaches to an adoptive cellular immunotherapy, sorted IE1 epitope-specific CD8-TM cells were found to be exceedingly protective upon adoptive transfer. Compared with CTLLs specific for the same epitope and of comparable avidity and TCR beta-chain variable region (Vbeta)-defined polyclonality, sorted CD8-TM cells proved to be superior by more than 2 orders of magnitude.     

Fire affects the occurrence of small mammals at distinct spatial scales in a neotropical savanna

Fire is a major disturbance event that affects biomes worldwide, altering vegetation structure and flora and fauna assemblages. Here, we investigated the effects of an extensive wildfire (~?6240 ha) on small mammal assemblages in savanna woodland (cerradão) at two spatial scales (meso- and macrohabitat) in a neotropical savanna (Brazilian Cerrado). At each spatial scale, we assessed relationships between habitat structure and small mammal species composition and abundance before and after the fire in four natural patches (one burned, three unburned) using partial redundancy analysis. From July 2009 to October 2013, we captured 1319 individuals belonging to 14 species. Our results showed that the fire had consequences for small mammal assemblage at both scales. After the fire, the burned patch differed from the other patches in vegetation attributes and species composition. At a fine scale, fire increased the herbaceous layer and decreased the litter layer and understory obstruction. On a larger scale, the main consequences of fire on vegetation structure were increased variation in litter depth, tree diameter, and distance to the nearest tree. We observed a relationship between mesohabitat structure and the abundance of species with different habitat requirements during the post-fire succession. Fire favored the invasion of generalist species from open Cerrado habitats (rodents Calomys tener, Calomys expulsus, Cerradomys scotti, and Necromys lasiurus) at the expense of more specialized forest species. Our results reinforce the relevance of incorporating multi-scale habitat heterogeneity in future studies assessing the effects of fire on wildlife.     

Beneficial effect of hot spring bathing on stress levels in Japanese macaques

The ability of animals to survive dramatic climates depends on their physiology, morphology and behaviour, but is often influenced by the configuration of their habitat. Along with autonomic responses, thermoregulatory behaviours, including postural adjustments, social aggregation, and use of trees for shelter, help individuals maintain homeostasis across climate variations. Japanese macaques (Macaca fuscata) are the world’s most northerly species of nonhuman primates and have adapted to extremely cold environments. Given that thermoregulatory stress can increase glucocorticoid concentrations in primates, we hypothesized that by using an available hot spring, Japanese macaques could gain protection against weather-induced cold stress during winter. We studied 12 adult female Japanese macaques living in Jigokudani Monkey Park, Japan, during the spring birth season (April to June) and winter mating season (October to December). We collected faecal samples for determination of faecal glucocorticoid (fGC) metabolite concentrations by enzyme immunoassay, as well as behavioural data to determine time spent in the hot springs, dominance rank, aggression rates, and affiliative behaviours. We used nonparametric statistics to examine seasonal changes in hot spring bathing, and the relationship between rank and air temperature on hot spring bathing. We used general linear mixed-effect models to examine factors impacting hormone concentrations. We found that Japanese macaques use hot spring bathing for thermoregulation during the winter. In the studied troop, the single hot spring is a restricted resource favoured by dominant females. High social rank had both costs and benefits: dominant females sustained high fGC levels, which were associated with high aggression rates in winter, but benefited by priority of access to the hot spring, which was associated with low fGC concentrations and therefore might help reduce energy expenditure and subsequent body heat loss. This unique habit of hot spring bathing by Japanese macaques illustrates how behavioural flexibility can help counter cold climate stress, with likely implications for reproduction and survival.     

Decreased PGC1-α levels and increased apoptotic protein signaling are associated with the maladaptive cardiac hypertrophy in hyperthyroidism

Hyperthyroidism can lead to the activation of proteins which are associated with inflammation, apoptosis, hypertrophy, and heart failure. This study aimed to explore the inflammatory and apoptotic proteins involved in the hyperthyroidism-induced cardiac hypertrophy establishment. Male Wistar rats were divided into control and hyperthyroid (12 mg/L L-thyroxine, in drinking water for 28 days) groups. The expression of inflammatory and apoptotic signaling proteins was quantified in the left ventricle by Western blot. Hyperthyroidism was confirmed by evaluation of T3 and T4 levels, as well as cardiac hypertrophy development. There was no change in the expression of HSP70, HIF1-α, TNF-α, MyD88, p-NFκB, NFκB, p-p38, and p38. Reduced expression of p53 and PGC1-α was associated with increased TLR4 and decreased IL-10 expression. Decreased Bcl-2 expression and increased Bax/Bcl-2 ratio were also observed. The results suggest that reduced PGC1-α and IL-10, and elevated TLR4 proteins expression could be involved with the diminished mitochondrial biogenesis and anti-inflammatory response, as well as cell death signaling, in the establishment of hyperthyroidism-induced maladaptive cardiac hypertrophy.     

Are pan traps colors complementary to sample community of potential pollinator insects?

The global initiatives of monitoring and conserving pollinators require worldwide assessments with comparable data sets collected through standardized methods. The use of pan traps is a passive method widely applied to sample flower visitors, standing out for its simplicity. Despite its wide use to sample pollinator diversity, the influence of color on trap efficiency is not well understood. The available studies are particularly scarce in the tropics and have generated divergent results. The main goal of the present study was to assess whether blue, yellow and white pan traps are complementary to sample Hymenoptera community. For this, we placed 49 sample units of blue, white and yellow pan traps in agricultural and natural (savanna-like) areas in Chapada Diamantina, Bahia, Brazil. We found that the species richness from blue and yellow pan traps were not significantly different, but both were significantly greater than the species richness from white pan traps. However, bees were significantly more attracted to the blue pan traps and wasps to the yellow ones; thus, color attractiveness was group-specific. Pan traps of different color showed low species composition overlap with 61 % of species collected exclusively in one of the three pan trap colors, and the species composition in the blue traps differed consistently from that in the traps of the other colors. In the article we discuss the implication of the results and defend the combined use of pan traps with different colors as a solution for the differential variable sample bias.     

Age,growth and mortality of the comber Serranus cabrilla (Linnaeus, 1758) in the Eastern Atlantic

Comber is an important bycatch species in the trawl fishery both in the Mediterranean and the Atlantic. Age and growth studies have been restricted to the eastern Mediterranean and no methodologies were ever applied to ensure accuracy and precision. In this study a total of 471 otoliths from specimens ranging from 12 to 26?cm were used. Precision indices showed a high level of agreement between readers and no evidence of systematic disagreement was obtained. To estimate fish growth and due to the absence of individuals smaller than 12?cm, three approaches were used to estimate the von Bertalanffy growth parameters: whole otolith readings, back calculation and a combination of both methodologies. Akaike’s information criterion suggests that the back calculation approach was the best one for describing comber growth: TL?=?25.26 [1?e^{(?0.21(t+1.72)}]. Natural mortality estimates varied between M?=?0.44 year^?1 and M?=?0.48 year^?1 while estimates of the fishing mortality varied between 0.12 year^?1 and 0.16 year^?1 and the exploitation rate between 0.20 and 0.27.     

Methionine and methionine sulfoxide alter parameters of oxidative stress in the liver of young rats: in vitro and in vivo studies

Hyperglycemia- and oxidative stress-induced modification of circulating lipoproteins is being increasingly recognized as an important pathogenetic factor for diabetic cardiovascular damages. This study was designed to investigate the impact of modified very low-density lipoprotein and high-density lipoprotein on phagocyte adhesion to endothelial cells and the involvement of scavenger receptor class B type 1 (SR-BI) in this process. Native lipoproteins were isolated by density gradient ultracentrifugation and in vitro glycoxidative or oxidative modification was performed in the presence of glucose or sodium hypochlorite, respectively. One hour co-incubation experiments with lipoproteins, freshly prepared polymorphonuclear leukocytes (PMN), and venous endothelial cells (HUVEC) were performed in the presence or absence of different scavenger receptors and signal transduction inhibitors. PMN adhesion to HUVEC was quantified fluorimetrically. We demonstrated that oxidized and glycoxidized lipoproteins promote adhesion of PMN to HUVEC from 1.5- to 2.5-fold with oxidized lipoproteins having the greatest effect. Treatment with the highly specific SR-BI inhibitor, BLT-1 produced substantial reduction of lipoprotein-induced adhesion to endothelial cells. Native and modified lipoproteins recruited extracellular signal-regulated kinase (ERK 1/2), p38 mitogen-activated protein kinase, and Janus kinase 2 as downstream signaling pathways for adhesion. From this study, it could be concluded that modification of lipoproteins plays a crucial role in atherosclerotic progression and SR-BI may be considered as a potential therapeutic target for the prevention of diabetic cardiovascular complications.