Carbon-13 nuclear magnetic resonance analysis of [1-13C]glucose metabolism in Crithidia fasciculata. Evidence of CO2 fixation by phosphoenolpyruvate carboxykinase

The non-invasive technique of 13C nuclear magnetic resonance was applied to study glucose metabolism in vivo in the insect parasite Crithidia fasciculata. It was found that under anaerobic conditions [1-13C]glucose underwent a glycolytic pathway whose main metabolic products were identified as [2-13C]ethanol, [2-13C]succinate and [1,3-13C2]glycerol. These metabolites were excreted by C. fasciculata into the incubation medium, while in the cells [3-13C]phosphoenolpyruvate was also detected in addition to the aforementioned compounds. The C3 acid is apparently the acceptor of the primary CO2 fixation reaction, which leads in Trypanosomatids to the synthesis of succinate. By addition of sodium bicarbonate to the incubation mixture L-[3-13C]malate was detected among the excretion products, while the ethanol:succinate ratio of 2.0 in the absence of bicarbonate changed to a ratio of 0.6 in the presence of the latter. This was due to a shift of the balance between carboxylation of phosphoenolpyruvate, leading to succinate, and pyruvate decarboxylation leading to ethanol. The addition of 25% 2H2O to the incubation mixture led to the formation of [2-13C, 2-2H]ethanol derived from the prior incorporation of 2H+ into pyruvate in the reactions mediated by either pyruvate kinase or malic enzyme. However, no 2H+ incorporation into L-malate was detected, excluding the possibility that the latter was formed by carboxylation of pyruvate, and lending support to the idea that L-malate results from the carboxylation of phosphoenolpyruvate to oxaloacetate by phosphoenolpyruvate carboxykinase. The formation of [2-13C, 2-2H]-succinate under the same conditions reflected the uptake of 2H+ during the reduction of fumarate. When the incubations were carried out in the presence of 100% 2H2O, several [1-13C, 1-2H]ethanol species were detected, as well as [2-13C, 2-2H]malate and [1,3-13C2, 1,3-2H2]glycerol. The former deuterated compounds reflect the existence of NAD2H species when the incubations were carried out in 100% 2H2O, while the incorporation of 2H+ into [1,3-13C2]glycerol must be attributed to the phosphoglucose-isomerase-mediated reaction during glycolysis.     

Context‐dependency in carnivore co‐occurrence across a multi‐use conservation landscape

Carnivore intraguild dynamics depend on a complex interplay of environmental affinities and interspecific interactions. Context‐dependency is commonly expected with varying suites of interacting species and environmental conditions but seldom empirically described. In South Africa, decentralized approaches to conservation and the resulting multi‐tenure conservation landscapes have markedly altered the environmental stage that shapes the structure of local carnivore assemblages. We explored assemblage‐wide patterns of carnivore spatial (residual occupancy probability) and temporal (diel activity overlap) co‐occurrence across three adjacent wildlife‐oriented management contexts—a provincial protected area, a private ecotourism reserve, and commercial game ranches. We found that carnivores were generally distributed independently across space, but existing spatial dependencies were context‐specific. Spatial overlap was most common in the protected area, where species occur at higher relative abundances, and in game ranches, where predator persecution presumably narrows the scope for spatial asymmetries. In the private reserve, spatial co‐occurrence patterns were more heterogeneous but did not follow a dominance hierarchy associated with higher apex predator densities. Pair‐specific variability suggests that subordinate carnivores may alternate between pre‐emptive behavioral strategies and fine‐scale co‐occurrence with dominant competitors. Consistency in species‐pairs diel activity asynchrony suggested that temporal overlap patterns in our study areas mostly depend on species'' endogenous clock rather than the local context. Collectively, our research highlights the complexity and context‐dependency of guild‐level implications of current management and conservation paradigms; specifically, the unheeded potential for interventions to influence the local network of carnivore interactions with unknown population‐level and cascading effects.     

Cryptogenic hepatitis patients have a higher Bartonella sp.-DNA detection in blood and skin samples than patients with non-viral hepatitis of known cause

BackgroundThis study aimed to assess the prevalence of Bartonella sp.-DNA detection in blood and skin samples from patients with non-viral end-stage liver disease awaiting liver transplantation.Methodology/Principal findingsBlood samples and healthy skin fragments from 50 patients were tested using microbiological and molecular methods. Fifteen patients had cryptogenic hepatitis (CH) and 35 had alcoholic, drug-induced or autoimmune liver disease. DNA was extracted from whole blood and liquid culture samples, isolates, and skin fragments. Thirteen of the 50 patients (26%) had Bartonella henselae DNA detection in their blood (9/50) and/or skin (5/50) samples. Colonies were isolated in 3/50 (6%) and infection was detected in 7/50 (14%) of the 50 patients. B. henselae-DNA detection was more prevalent in patients with CH than in other patients (p = 0.040). Of 39 patients followed-up for at least two years, a higher mortality rate was observed among patients with CH infected with B. henselae (p = 0.039).Conclusions/SignificanceFurther studies assessing the role of B. henselae infection in the pathogenesis of hepatitis patients must be urgently conducted.     

Reduced superoxide dismutase activity in Palaemonetes argentinus (Decapoda, Palemonidae) infected by Probopyrus ringueleti (Isopoda, Bopyridae)

Cellular oxidative stress may promote damage or death in biological systems and may be caused by production of pro-oxidant molecules known as reactive oxygen species (ROS). The aim of this work was to analyze the activity of antioxidant enzymes (catalase [CAT], superoxide dismutase [SOD] and glutathione peroxidase [GPx]) in the shrimp Palaemonetes argentinus Nobili, 1901 infected by Probopyrus ringueleti (Verdi & Schuldt, 1987), a gill chamber parasite known for its capacity to cause host metabolic changes, including changes in oxygen consumption rates. Infested and non-infested shrimp were collected in the Patos Lagoon estuary (southern Brasil), where the prevalence of the parasite may be as high as 70%. No significant differences were observed for either CAT or GPx activities. However, SOD activity was significantly reduced in infected shrimp, suggesting that bopyrid isopod respiratory impairment resulted in reduced SOD enzyme activity.     

Profile of total IgG, IgG1, IgG2, IgG3 and IgG4 levels in sera of patients with paracoccidioidomycosis: treatment follow-up using Mexo and rPb27 as antigens in an ELISA

The levels of total of IgG, IgG1, IgG2, IgG3 and IgG4 were evaluated in 54 patients with chronic paracoccidioidomycosis (PCM) before, during and after treatment using an enzyme-linked immunosorbent assay with Mexo and recombinant Pb27 (rPb27) as the antigens. Mexo was effective in distinguishing PCM patients from individuals in the negative control group (NC) based on total IgG and rPb27 performed worse than Mexo when these two groups were compared. IgG1, IgG2, IgG3 and IgG4 could not be used to clearly distinguish PCM patients from those in the NC group using either antigen. There was no clear relationship between antibody levels and the period of treatment. The majority of patients presented with decreased antibody levels during treatment, with no statistically significant differences among the different periods of treatment. Only IgG4 presented a negative correlation between its levels and clinical improvement during treatment. In total, 65% of untreated PCM patients showed reactivity against IgG4 when the Mexo antigen was used and this reactivity decreased over the course of treatment. There was a tendency towards decreasing antibody levels during treatment, but these antibody levels did not necessarily clear after the treatment was stopped. Mexo was useful for PCM diagnosis using total IgG; however, more studies are necessary before this antigen can be used in measuring the levels of total IgG and its subclasses for monitoring patients during treatment.     

High dietary salt decreases antioxidant defenses in the liver of fructose-fed insulin-resistant rats

In this study we investigated the hypothesis that a high-salt diet to hyperinsulinemic rats might impair antioxidant defense owing to its involvement in the activation of sodium reabsorption to lead to higher oxidative stress. Rats were fed a standard (CON), a high-salt (HS), or a high-fructose (HF) diet for 10 weeks after which, 50% of the animals belonging to the HF group were switched to a regimen of high-fructose and high-salt diet (HFS) for 10 more weeks, while the other groups were fed with their respective diets. Animals were then euthanized and their blood and liver were examined. Fasting plasma glucose was found to be significantly higher (approximately 50%) in fructose-fed rats than in the control and HS rats, whereas fat liver also differed in these animals, producing steatosis. Feeding fructose-fed rats with the high-salt diet triggered hyperinsulinemia and lowered insulin sensitivity, which led to increased levels of serum sodium compared to the HS group. This resulted in membrane perturbation, which in the presence of steatosis potentially enhanced hepatic lipid peroxidation, thereby decreasing the level of antioxidant defenses, as shown by GSH/GSSG ratio (HFS rats, 7.098±2.1 versus CON rats, 13.2±6.1) and superoxide dismutase (HFS rats, 2.1±0.05 versus CON rats, 2.3±0.1%), and catalase (HFS rats, 526.6±88.6 versus CON rats, 745.8±228.7 U/mg ptn) activities. Our results indicate that consumption of a salt-rich diet by insulin-resistant rats may lead to regulation of sodium reabsorption, worsening hepatic lipid peroxidation associated with impaired antioxidant defenses.     

Greater macroinvertebrate diversity and freshwater mussel density in meander margins of an Amazon river

Meanders are complex aquatic environments exhibiting different flow and sediment characteristics that influence macroinvertebrate distribution. Differences in macroinvertebrate composition, freshwater mussel density (mainly Castalia ambigua Lamarck, 1819) and habitat variables were investigated, using uni‐ and multivariate analyses, in two zones, the margin and thalweg, of the meanders of a 7‐km stretch of a morphologically unaltered tropical alluvial lowland river. Clear differences were found between meander zones, with greater taxonomic and functional diversity of macroinvertebrates and greater freshwater mussel densities in the meander margin. A total of 12 families of macroinvertebrates were exclusive to the meander margin, and macroinvertebrate indicators of zones were Philopotamidae and Leptoceridae (meander margin) and Chironomidae and Elmidae (meander thalweg). In the meander margin, there were no differences in macroinvertebrate abundance, and taxonomic and functional group diversity among areas with low, medium and high mussel density. Macroinvertebrate abundance did not vary between zones, but in both, abundance was associated with lower pH and higher electrical conductivity. Low shear velocity, which stabilises the river bed, high organic matter content and percentage silt in the meander margin were associated with higher taxonomic and functional macroinvertebrate diversity, as well as the presence of freshwater mussels. Natural unaltered meanders are spatially heterogeneous in both habitat and biodiversity and, similar to restored meanders, support greater macroinvertebrate taxonomic and functional diversity, as well as maintain freshwater mussel beds, mainly by stabilising the river bed. The meander margin may also be important for attracting mussel host fish that feed on macroinvertebrates and aid mussel dispersal. Thus, the meander margins, and other similar riparian habitats with low shear velocity, for example, side channels and pools, have potentially high conservation value and should be afforded due protection.     

Functional Recovery Occurs Even After Partial Remyelination of Axon-Meshed Median and Ulnar Nerves in Mice

Upper limb nerve injuries are common, and their treatment poses a challenge for physicians and surgeons. Experimental models help in minimum exploration of the functional characteristics of peripheral nerve injuries of forelimbs. This study was conducted to characterize the functional recovery (1, 3, 7, 10, 14, and 21 days) after median and ulnar nerve crush in mice and analyze the histological and biochemical markers of nerve regeneration (after 21 days). Sensory–functional impairments appeared after 1 day. The peripheral nerve morphology, the nerve structure, and the density of myelin proteins [myelin protein zero (P0) and peripheral myelin protein 22 (PMP22)] were analyzed after 21 days. Cold allodynia and fine motor coordination recovery occurred on the 10th day, and grip strength recovery was observed on the 14th day after injury. After 21 days, there was partial myelin sheath recovery. PMP22 recovery was complete, whereas P0 recovery was not. Results suggest that there is complete functional recovery even with partial remyelination of median and ulnar nerves in mice.