Methyl-group donors cannot prevent apoptotic death of rat hepatocytes induced by choline-deficiency

Choline-deficiency causes liver cells to die by apoptosis, and it has not been clear whether the effects of choline-deficiency are mediated by methyl-deficiency or by lack of choline moieties. SV40 immortalized CWSV-1 hepatocytes were cultivated in media that were choline-sufficient, choline-deficient, choline-deficient with methyl-donors (betaine or methionine), or choline-deficient with extra folate/vitamin B₁₂. Choline-deficient CWSV-1 hepatocytes were not methyl-deficient as they had increased intracellular S-adenosylmethionine concentrations (132% of control; P < 0.01). Despite increased phosphatidylcholine synthesis via sequential methylation of phosphatidylethanolamine, choline-deficient hepatocytes had significantly decreased (P < 0.01) intracellular concentrations of choline (20% of control), phosphocholine (6% of control), glycerophosphocholine (15% of control), and phosphatidylcholine (55% of control). Methyl-supplementation in choline-deficiency enhanced intracellular methyl-group availability, but did not correct choline-deficiency induced abnormalities in either choline metabolite or phospholipid content in hepatocytes. Methyl-supplemented, choline-deficient cells died by apoptosis. In a rat study, 2 weeks of a choline-deficient diet supplemented with betaine did not prevent the occurrence of fatty liver and the increased DNA strand breakage induced by choline-deficiency. Though dietary supplementation with betaine restored hepatic betaine concentration and increased hepatic S-adenosylmethionine/S-adenosylhomocysteine ratio, it did not correct depleted choline (15% of control), phosphocholine (6% control), or phosphatidylcholine (48% of control) concentrations in deficient livers. These data show that decreased intracellular choline and/or choline metabolite concentrations, and not methyl deficiency, are associated with apoptotic death of hepatocytes. J. Cell. Biochem, 64:196–208. © 1997 Wiley-Liss, Inc.     

Dengue and Zika virus infection patterns vary among Aedes aegypti field populations from Belo Horizonte,a Brazilian endemic city

Dengue virus (DENV) and Zika virus (ZIKV) belong to the same viral family, the Flaviviridae. They cause recurring threats to the public health systems of tropical countries such as Brazil. The primary Brazilian vector of both viruses is the mosquito Aedes aegypti. After the mosquito ingests a blood meal from an infected person, the viruses infect and replicate in the midgut, disseminate to secondary tissues and reach the salivary gland (SG), where they are ready to be transmitted to a vertebrate host. It is thought that the intrinsic discrepancies among mosquitoes could affect their ability to deal with viral infections. This study confirms that the DENV and ZIKV infection patterns of nine Ae. aegypti field populations found in geographically separate health districts of an endemic Brazilian city vary. We analyzed the infection rate, disseminated infection, vector competence, and viral load through quantitative PCR. Mosquitoes were challenged using the membrane-feeding assay technique and were tested seven and fourteen days post-infection (early and late infection phases, respectively). The infection responses varied among the Ae. aegypti populations for both flaviviruses in the two infection phases. There was no similarity between DENV and ZIKV vector competencies or viral loads. According to the results of our study, the risk of viral transmission overtime after infection either increases or remains unaltered in ZIKV infected vectors. However, the risk may increase, decrease, or remain unaltered in DENV-infected vectors depending on the mosquito population. For both flaviviruses, the viral load persisted in the body even until the late infection phase. In contrast to DENV, the ZIKV accumulated in the SG over time in all the mosquito populations. These findings are novel and may help direct the development of control strategies to fight dengue and Zika outbreaks in endemic regions, and provide a warning about the importance of understanding mosquito responses to arboviral infections.     

Autophagy in Spinocerebellar ataxia type 2, a dysregulated pathway,and a target for therapy

Spinocerebellar ataxia type 2 (SCA2) is an incurable and genetic neurodegenerative disorder. The disease is characterized by progressive degeneration of several brain regions, resulting in severe motor and non-motor clinical manifestations. The mutation causing SCA2 disease is an abnormal expansion of CAG trinucleotide repeats in the ATXN2 gene, leading to a toxic expanded polyglutamine segment in the translated ataxin-2 protein. While the genetic cause is well established, the exact mechanisms behind neuronal death induced by mutant ataxin-2 are not yet completely understood. Thus, the goal of this study is to investigate the role of autophagy in SCA2 pathogenesis and investigate its suitability as a target for therapeutic intervention. For that, we developed and characterized a new striatal lentiviral mouse model that resembled several neuropathological hallmarks observed in SCA2 disease, including formation of aggregates, neuronal marker loss, cell death and neuroinflammation. In this new model, we analyzed autophagic markers, which were also analyzed in a SCA2 cellular model and in human post-mortem brain samples. Our results showed altered levels of SQSTM1 and LC3B in cells and tissues expressing mutant ataxin-2. Moreover, an abnormal accumulation of these markers was detected in SCA2 patients’ striatum and cerebellum. Importantly, the molecular activation of autophagy, using the compound cordycepin, mitigated the phenotypic alterations observed in disease models. Overall, our study suggests an important role for autophagy in the context of SCA2 pathology, proposing that targeting this pathway could be a potential target to treat SCA2 patients.Subject terms: Diseases of the nervous system, Molecular neuroscience     

The signatures of organellar calcium

Recent insights about the transport mechanisms involved in the in and out of calcium ions in plant organelles, and their role in the regulation of cytosolic calcium homeostasis in different signaling pathways.

The transport of Ca²⁺ across the membranes of subcellular compartments contributes to cytosolic Ca²⁺ homeostasis as well as environmental and developmental responses.     

The discovery of IDX21437: Design,synthesis and antiviral evaluation of 2′-α-chloro-2′-β-C-methyl branched uridine pronucleotides as potent liver-targeted HCV polymerase inhibitors

Herein we describe the discovery of IDX21437 35b, a novel R_P d-aminoacid-based phosphoramidate prodrug of 2′-α-chloro-2′-β-C-methyluridine monophosphate. Its corresponding triphosphate 6 is a potent inhibitor of the HCV NS5B RNA-dependent RNA polymerase (RdRp). Despite showing very weak activity in the in vitro Huh-7 cell based HCV replicon assay, 35b demonstrated high levels of active triphosphate 6 in mouse liver and human hepatocytes. A biochemical study revealed that the metabolism of 35b was mainly attributed to carboxyesterase 1 (CES1), an enzyme which is underexpressed in HCV Huh-7-derived replicon cells. Furthermore, due to its metabolic activation, 35b was efficiently processed in liver cells compared to other cell types, including human cardiomyocytes. The selected R_P diastereoisomeric configuration of 35b was assigned by X-ray structural determination. 35b is currently in Phase II clinical trials for the treatment of HCV infection.     

The defensive behavioral patterns of captive white-lipped and collared peccary (Mammalia,Tayassuidae): an approach for conservation of the species

Defensive behavioral patterns in response to human-induced rapid environmental change can affect animals’ fitness and may play a role in species conservation status. To test this hypothesis, we compared the risk assessment and defensive behavioral responses of captive white-lipped peccary (WLP; Tayassu pecari) and collared peccary (CP; Pecari tajacu), which retain different conservation status; WLP are considered vulnerable and CP of least concern. We used an adapted paradigm of the mouse defense test battery (MDTB) comprising four consecutive tests. Two of these tests simulated a novel environment, while the other two stimulated the expression of defensive behavioral patterns. Besides differences in risk assessment and defensive threat/attack behavioral patterns between species, we compared flight initiation distance, flight speed, and plasma glucocorticoid concentrations. When facing a novel environment and risk challenges from humans’ predator-like cues, the white-lipped peccary showed more exploratory and defensive threat/attack behavioral patterns, shorter flight initiation distances, and lower flight speeds, whereas the collared peccaries showed more cautious and retreat patterns, longer flight initiation distances, and higher flight speeds. There were also correlations between physiological and behavioral parameters. We confirmed our hypothesis that the collared peccary’s cautiousness may help to prevent a decrease in its population, while the white-lipped peccary’s exploratory and confrontational behavioral patterns in overhunted areas, together with other simultaneous factors as forest fragmentation, might contribute to placing this species in the vulnerable category.     

Chemical sex recognition in the harvestman <Emphasis Type="Italic">Discocyrtus prospicuus</Emphasis> (Arachnida: Opiliones)

Several arachnid species use chemicals to detect sexual partners. In harvestmen, there are evidences that chemicals may play a role in intraspecific communication. Using the behavior of Discocyrtus prospicuus (Holmberg 1876), whose males expose the penis to females before they engage in mating posture, we tested if males detect females by contact chemoreception (chemicals left on the substrate) and if males detect females by olfaction. First, we exposed males to three experimental groups, where males had to choose between two substrates: female chemicals/blank control; male chemicals/blank control; female/male chemicals. Then, we gave males access to volatiles of males, females, and control simultaneously. We predicted that males would expose the penis when approaching volatiles and chemicals deposited on the substrate by females. We also tested if males spent more time close to the source of female volatiles and on the substrate with female chemicals and if males tapped the substrate with female chemicals for more time than the others. Finally, we put males and females together to observe if males would expose the penis upon touching the female’s cuticle. Most of our predictions were not supported, though males did tap for more time when exposed to female cues instead of male cues and exposed the penis in 70% of the observations when interacting with the female but only after touching her. Our data does not support olfaction as a way to detect females and corroborate the idea that contact chemicals, either on the substrate or on female’s cuticle, play an important role in the detection and recognition of the opposite sex. This is the first evidence in harvestmen that males may react differently to female/male chemicals.     

Local lithological drivers of post-fire vegetation recovery and implications for fire-prone regions

Local ecosystem resilience to fire disturbance can be influenced by multiple factors, from topography and climate, to fire history and pre-fire structure of biotic communities. Here we investigated the factors affecting post-fire recovery of scrub vegetation in areas under Mediterranean climate affected by frequent fires. We hypothesized that, under comparable climatic and topographic conditions, geological factors (with bedrock type as a proxy) would be at least as important as fire history in explaining patterns of post-fire recovery. We surveyed scrub vegetation in a mountain study area in Portugal, using a stratified random sampling scheme, with fire frequency, time since last fire, and bedrock type (granite vs. schist) as stratifying layers. Based on vegetation and plant community data from 40 plots, we analyzed total species richness and composition, and the relative abundance of functional groups defined on the basis of general (non fire-specific) life-history traits. We found that, at a local scale, lithology can override fire history in determining post-fire recovery. Vegetation plots on granite exhibited a considerable development of tall scrubs and higher values of total species richness. They also hosted higher numbers of animal-dispersed woody species, of trees and tall scrubs, of woody deciduous species, and of forest, edge and tall scrub species. Differences in the post-fire development of scrub vegetation and in the functional profile of plant communities highlight the need to consider local geological diversity when establishing priorities for post-fire active restoration under scenarios of limited resources.