The role of l-arginine-nitric oxide pathway in bacterial translocation

This study investigated the nitric oxide (NO) role as a mediator of arginine on bacterial translocation (BT) and gut damage in mice after intestinal obstruction (IO). The effects of pretreatment with arginine with or without NO inhibition on the systemic and local immunological response were also assessed. Mice were categorized into four groups. Group ARG received chow containing 2 % arginine, while group ARG + l-NAME received the same diet plus l-NAME (N-nitro-l-arginine methyl ester) by gavage. The IO and Sham groups were fed standard chow. After 7 days, animals were gavaged with radiolabeled Escherichia coli, anesthetized and subjected to IO, except the Sham group. Animals were euthanized after 18 h, and BT was evaluated in the mesenteric lymph nodes, blood, liver, spleen and lungs. In another experiment, the intestinal injury was assessed regarding intestinal permeability and ileum histological analyses. Intestinal secretory immunoglobulin A (sIgA) levels, serum IFN-γ and IL-10 cytokines were assessed. Arginine reduced BT, but NO inhibition enhanced BT compared with the ARG group (p < 0.05). Intestinal permeability in the ARG and ARG + l-NAME groups was similar but decreased when compared with the IO group (p < 0.05). Histological preservation was observed. Arginine treatment increased IL-10 and sIgA levels when compared with the Sham and IO groups (p < 0.05). The cytokines and sIgA concentrations were similar in the ARG + l-NAME and Sham groups. Arginine appeared to reduce BT and its effects on the modulation of cytokines and secretory IgA in mice after IO are mediated by NO production.     

Importance of the interaction protein–protein of the CaM–PDE1A and CaM–MLCK complexes in the development of new anti‐CaM drugs

Protein–protein interactions play central roles in physiological and pathological processes. The bases of the mechanisms of drug action are relevant to the discovery of new therapeutic targets. This work focuses on understanding the interactions in protein–protein–ligands complexes, using proteins calmodulin (CaM), human calcium/calmodulin‐dependent 3′,5′‐cyclic nucleotide phosphodiesterase 1A active human (PDE1A), and myosin light chain kinase (MLCK) and ligands αII–spectrin peptide (αII–spec), and two inhibitors of CaM (chlorpromazine (CPZ) and malbrancheamide (MBC)). The interaction was monitored with a fluorescent biosensor of CaM (hCaM M124C–mBBr). The results showed changes in the affinity of CPZ and MBC depending on the CaM–protein complex under analysis. For the Ca²⁺–CaM, Ca²⁺–CaM–PDE1A, and Ca²⁺–CaM–MLCK complexes, CPZ apparent dissociation constants (K_ds) were 1.11, 0.28, and 0.55 μM, respectively; and for MBC K_ds were 1.43, 1.10, and 0.61 μM, respectively. In competition experiments the addition of calmodulin binding peptide 1 (αII–spec) to Ca²⁺–hCaM M124C–mBBr quenched the fluorescence (K_d = 2.55 ± 1.75 pM) and the later addition of MBC (up to 16 μM) did not affect the fluorescent signal. Instead, the additions of αII–spec to a preformed Ca²⁺–hCaM M124C–mBBr–MBC complex modified the fluorescent signal. However, MBC was able to displace the PDE1A and MLCK from its complex with Ca²⁺–CaM. In addition, docking studies were performed for all complexes with both ligands showing an excellent correlation with experimental data. These experiments may help to explain why in vivo many CaM drugs target prefer only a subset of the Ca²⁺–CaM regulated proteins and adds to the understanding of molecular interactions between protein complexes and small ligands. Copyright © 2013 John Wiley & Sons, Ltd.     

Is forest certification targeting areas of high biodiversity in cork oak savannas?

Over the last four decades the world has been losing biodiversity at an alarming rate despite the increasing number of protected areas (PAs). Certified forest management may complement the role of PAs in protecting biodiversity. Forest certification aims to promote sustainable forest management and to maintain or enhance the conservation value of certified forests. The area of forest under certified forest management has grown quickly over the past decade. Forest Stewardship Council (FSC) certification, for example, currently covers 148 million hectares, i.e., 3.7 % of the world’s forests. In spite of such increase there is, however, a dearth of information on how forest certification is related to biodiversity. In this study we assessed if FSC certification is being applied in high biodiversity areas in cork oak savannas in Portugal by comparing biodiversity values of certified and non-certified areas for birds, reptiles and amphibians. We calculated the relative species richness and irreplaceability value for each group of species in certified and non-certified areas and compared them using randomization tests. The biodiversity value of certified areas was not significantly greater than that of non-certified areas. Since FSC certification is expanding quickly in cork oak savannas it is important to consider the biodiversity value of these areas during this process. Prioritizing areas of high biodiversity value would enhance the conservation value of forest certification and facilitate integrating certification with other conservation initiatives.     

Richness and composition of Calliphoridae in an Atlantic Forest fragment: implication for the use of dipteran species as bioindicators

Studies measuring the damage in degraded environments have increased, and it is necessary to obtain reliable biological and ecological indicators for the recovery of such degraded environments and subsequent preservation. This study aimed to conduct a survey of local insects and evaluate their utility in monitoring forest restoration in a degraded area in the Atlantic Forest biome in Ribeirao Grande, State of São Paulo, Brazil. Collections with Shuey traps were conducted from 2006 to 2011 in four distinct forest fragments characterized by environmental impacts, human actions, and phytophysiognomic profiles: Forest (FO), a preserved area with native plant species; “Capoeira” (CA), an area in the natural regeneration process; Planting (PL), an area reforested with native plants; and Pasture (PA), an area with only shrubs. A total of 2,456 specimens of Calliphoridae (Diptera) were collected. Mesembrinella bellardiana was the most abundant (n = 1,884) and dominant species in all environments sampled. The relative abundance of M. bellardiana in the most preserved environment (60.2 % FO, 24.4 % CA, 8.6 % PL, and 6.7 % PA) and other ecological parameters showed that it could be a bioindicator species, i.e., data on its presence or absence directly reflected the status of local preservation. Information on Neotropical dipterans associated with the forested environment is very scarce in the literature. To our knowledge, this is the first study describing the occurrence of Calliphora lopesi (Calliphoridae) in the State of São Paulo.     

Disturbances, elevation, topography and spatial proximity drive vegetation patterns along an altitudinal gradient of a top biodiversity hotspot

The correlation between vegetation patterns (species distribution and richness) and altitudinal variation has been widely reported for tropical forests, thereby providing theoretical basis for biodiversity conservation. However, this relationship may have been oversimplified, as many other factors may influence vegetation patterns, such as disturbances, topography and geographic distance. Considering these other factors, our primary question was: is there a vegetation pattern associated with substantial altitudinal variation (10–1,093 m a.s.l.) in the Atlantic Rainforest—a top hotspot for biodiversity conservation—and, if so, what are the main factors driving this pattern? We addressed this question by sampling 11 1-ha plots, applying multivariate methods, correlations and variance partitioning. The Restinga (forest on sandbanks along the coastal plains of Brazil) and a lowland area that was selectively logged 40 years ago were floristically isolated from the other plots. The maximum species richness (>200 spp. per hectare) occurred at approximately 350 m a.s.l. (submontane forest). Gaps, multiple stemmed trees, average elevation and the standard deviation of the slope significantly affected the vegetation pattern. Spatial proximity also influenced the vegetation pattern as a structuring environmental variable or via dispersal constraints. Our results clarify, for the first time, the key variables that drive species distribution and richness across a large altitudinal range within the Atlantic Rainforest.     

Molecular mechanisms beyond glucose transport in diabetes-related male infertility

Diabetes mellitus (DM) is one of the greatest public health threats in modern societies. Although during a few years it was suggested that DM had no significant effect in male reproductive function, this view has been challenged in recent years. The increasing incidence of DM worldwide will inevitably result in a higher prevalence of this pathology in men of reproductive age and subfertility or infertility associated with DM is expected to dramatically rise in upcoming years. From a clinical perspective, the evaluation of semen parameters, as well as spermatozoa deoxyribonucleic acid (DNA) integrity, are often studied due to their direct implications in natural and assisted conception. Nevertheless, recent studies based on the molecular mechanisms beyond glucose transport in testicular cells provide new insights in DM-induced alterations in male reproductive health. Testicular cells have their own glucose sensing machinery that react to hormonal fluctuations and have several mechanisms to counteract hyper- and hypoglycemic events. Moreover, the metabolic cooperation between testicular cells is crucial for normal spermatogenesis. Sertoli cells (SCs), which are the main components of blood–testis barrier, are not only responsible for the physical support of germ cells but also for lactate production that is then metabolized by the developing germ cells. Any alteration in this tied metabolic cooperation may have a dramatic consequence in male fertility potential. Therefore, we present an overview of the clinical significance of DM in the male reproductive health with emphasis on the molecular mechanisms beyond glucose fluctuation and transport in testicular cells.     

Diurnal Variation of Plasmatic Melatonin,Corticosterone and Variation of General Activity in Pigeons under Light-Dark Cycle and Constant Light

This work analyzed the diurnal variation of general activity and plasmatic levels of melatonin and corticosterone in pigeons submitted to a 12:00:12:00 h light-dark cycle (lights on at 6:00 a.m.) or to constant light. In both conditions pigeons were observed in 5-min sessions at times 03:00, 06:00, 09:00, 12:00, 15:00, 18:00, 21:00 and 24:00 h during two successive days. Behavior was video taped in the home cages for posterior categorization and quantification. Radioimmunoassays were used to evaluate plasmatic levels of melatonin and corticosterone. Blood samples were obtained at the times of behavioral observation. In the light-dark condition the results showed day-night variation of general activity (p &lt; 0.001) and a robust diurnal rhythm of plasmatic melatonin (p &lt; 0.001). Both of these variations as well as the oscillatory secretion of corticosterone disappeared under constant light condition. The parallel changes in general activity and blunting of melatonin rhythm secretion in constant light condition agree with previous evidences that melatonin may regulate behavioral oscillations in the pigeon. The present data are related to the proposition that the timing system in pigeons may involve neuroendocrine relations characterized by interactions between blood born signalization by melatonin and corticosterone.     

Diurnal Variation in Oral Temperature,Sleepiness, and Performance of High School Girls

Evaluation of rhythmic fluctuations of physical and mental variables should be of special significance for the understanding of students' performance and setting the schedules of school activities. The present study investigated the pattern of diurnal variation in oral temperature, sleepiness and performance of a group of adolescents undergoing a daytime school schedule. Eighteen girls (mean age 16 years-old), who attended the same class from 0715h to 1645h, were tested on seven days. They measured their oral temperature, quantified their sleepiness level by means of a visual analogue scale, and completed the following tests: letter cancellation test, addition test, and a simple motor task. One-way ANOVA statistics for repeated measures was used in order to verify the effect of test time on oral temperature, sleepiness and performance. Possible correlations between the level of sleepiness and performance were investigated by means of Spearman rank correlation. The results revealed significant time of day effect on all variables, except for the number of addition errors. Oral temperature values showed an increase from morning to afternoon. Letter cancellation, motor task and addition scores increased from early morning to late afternoon, showing rapid fluctuations throughout the day. Sleepiness level was negatively correlated with letter cancellation scores during the first three tests of the day. In agreement with other work, the diurnal variation of oral temperature, letter cancellation and addition test showed an improvement as the day progressed. Sleepiness, on the other hand, decreased throughout the day, with the highest level associated with the first test of the day, suggesting a circadian pattern of variation rather than a cumulative effect due to school activities.     

Drosophila ref(2)P is required for the parkin-mediated suppression of mitochondrial dysfunction in pink1 mutants

Autophagy is a critical regulator of organellar homeostasis, particularly of mitochondria. Upon the loss of membrane potential, dysfunctional mitochondria are selectively removed by autophagy through recruitment of the E3 ligase Parkin by the PTEN-induced kinase 1 (PINK1) and subsequent ubiquitination of mitochondrial membrane proteins. Mammalian sequestrome-1 (p62/SQSTM1) is an autophagy adaptor, which has been proposed to shuttle ubiquitinated cargo for autophagic degradation downstream of Parkin. Here, we show that loss of ref(2)P, the Drosophila orthologue of mammalian P62, results in abnormalities, including mitochondrial defects and an accumulation of mitochondrial DNA with heteroplasmic mutations, correlated with locomotor defects. Furthermore, we show that expression of Ref(2)P is able to ameliorate the defects caused by loss of Pink1 and that this depends on the presence of functional Parkin. Finally, we show that both the PB1 and UBA domains of Ref(2)P are crucial for mitochondrial clustering. We conclude that Ref(2)P is a crucial downstream effector of a pathway involving Pink1 and Parkin and is responsible for the maintenance of a viable pool of cellular mitochondria by promoting their aggregation and autophagic clearance.