Long-Term Behavioral Programming Induced by Peripuberty Stress in Rats Is Accompanied by GABAergic-Related Alterations in the Amygdala

Stress during childhood and adolescence is a risk factor for psychopathology. Alterations in γ-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the brain, have been found following stress exposure and fear experiences and are often implicated in anxiety and mood disorders. Abnormal amygdala functioning has also been detected following stress exposure and is also implicated in anxiety and social disorders. However, the amygdala is not a unitary structure; it includes several nuclei with different functions and little is known on the potential differences the impact of early life stress may have on this system within different amygdaloid nuclei. We aimed here to evaluate potential regional differences in the expression of GABAergic-related markers across several amygdaloid nuclei in adult rats subjected to a peripuberty stress protocol that leads to enhanced basal amygdala activity and psychopathological behaviors. More specifically, we investigated the protein expression levels of glutamic acid decarboxylase (GAD; the principal synthesizing enzyme of GABA) and of GABA-A receptor subunits α2 and α3. We found reduced GAD and GABA-A α3, but not α2, subunit protein levels throughout all the amygdala nuclei examined (lateral, basolateral, basomedial, medial and central) and increased anxiety-like behaviors and reduced sociability in peripubertally stressed animals. Our results identify an enduring inhibition of the GABAergic system across the amygdala following exposure to early adversity. They also highlight the suitability of the peripuberty stress model to investigate the link between treatments targeting the dysfunctional GABAergic system in specific amygdala nuclei and recovery of specific stress-induced behavioral dysfunctions.     

Direct Comparison of the Efficacy and Safety of Oral Treatments with Oleylphosphocholine (OlPC) and Miltefosine in a Mouse Model of L. major Cutaneous Leishmaniasis

Background

Cutaneous leishmaniasis (CL) represents a range of skin diseases caused by infection with Leishmania parasites and associated with tissue inflammation and skin ulceration. CL is clinically widespread in both the Old and New World but lacks treatments that are well tolerated, effective and inexpensive. Oleylphosphocholine (OlPC) is a new orally bioavailable drug of the alkylphosphocholine family with potent antileishmanial activity against a broad range of Leishmania species/strains.

Methodology/principal findings

The potential of OlPC against Old World CL was evaluated in a mouse model of Leishmania (L.) major infection in BALB/c mice. Initial dose-response experiments showed that an oral daily dose of 40 mg/kg of OlPC was needed to impact time to cure and lesion sizes. This dose was then used to directly compare the efficacy of OlPC to the efficacy of the antileishmanial drugs miltefosine (40 mg/kg/day), fluconazole (160 mg/kg/day) and amphotericin B (25 mg/kg/day). OlPC, miltefosine and fluconazole were given orally for 21 days while amphotericin B was administered intraperitoneally for 10 days. Ulcer sizes and animal weights were followed up on a weekly basis and parasitemia was determined by means of a real-time in vivo imaging system which detects luminescence emitted from luciferase-expressing infecting L. major parasites. Amphotericin B and OlPC showed excellent efficacy against L. major lesions in terms of reduction of parasitic loads and by inducing complete healing of established lesions. In contrast, treatment with miltefosine did not significantly affect parasitemia and lesion sizes, while fluconazole was completely ineffective at the dose regimen tested.

Conclusions/Significance

Given the data showing the outstanding efficacy and tolerability of OlPC, our results suggest that OlPC is a promising new drug candidate to improve and simplify current clinical management of L. major CL.     

Seed-competent tau monomer initiates pathology in a tauopathy mouse model

Tau aggregation into ordered assemblies causes neurodegenerative tauopathies. We previously reported that tau monomer exists in either inert (M_i) or seed-competent (M_s) conformational ensembles and that M_s encodes strains, that is, unique, self-replicating, biologically active assemblies. It is unknown if disease begins with M_s formation followed by fibril assembly or if M_s derives from fibrils and is therefore an epiphenomenon. Here, we studied a tauopathy mouse model (PS19) that expresses full-length mutant human (1N4R) tau (P301S). Insoluble tau seeding activity appeared at 2 months of age and insoluble tau protein assemblies by immunoblot at 3 months. Tau monomer from mice aged 1 to 6 weeks, purified using size-exclusion chromatography, contained soluble seeding activity at 4 weeks, before insoluble material or larger assemblies were observed, with assemblies ranging from n = 1 to 3 tau units. By 5 to 6 weeks, large soluble assemblies had formed. This indicated that the first detectable pathological forms of tau were in fact M_s. We next examined posttranslational modifications of tau monomer from 1 to 6 weeks. We detected no phosphorylation unique to M_s in PS19 or human Alzheimer’s disease brains. We conclude that tauopathy begins with formation of the M_s monomer, whose activity is phosphorylation independent. M_s then self assembles to form oligomers before it forms insoluble fibrils. The conversion of tau monomer from M_i to M_s thus constitutes the first detectable step in the initiation of tauopathy in this mouse model, with obvious implications for the origins of tauopathy in humans.     

The role of NCAM in auditory fear conditioning and its modulation by stress: a focus on the amygdala

Chronic stress in rodents was shown to induce structural shrinkage and functional alterations in the hippocampus that were linked to spatial memory impairments. Effects of chronic stress on the amygdala have been linked to a facilitation of fear conditioning. Although the underlying molecular mechanisms are still poorly understood, increasing evidence highlights the neural cell adhesion molecule (NCAM) as an important molecular mediator of stress‐induced structural and functional alterations. In this study, we investigated whether altered NCAM expression levels in the amygdala might be related to stress‐induced enhancement of auditory fear conditioning and anxiety‐like behavior. In adult C57BL/6J wild‐type mice, chronic unpredictable stress resulted in an isoform‐specific increase of NCAM expression (NCAM‐140 and NCAM‐180) in the amygdala, as well as enhanced auditory fear conditioning and anxiety‐like behavior. Strikingly, forebrain‐specific conditional NCAM‐deficient mice (NCAM‐floxed mice that express the cre‐recombinase under the control of the promoter of the α‐subunit of the calcium‐calmodulin‐dependent protein kinase II), whose amygdala NCAM expression levels are reduced, displayed impaired auditory fear conditioning which was not altered following chronic stress exposure. Likewise, chronic stress in these conditional NCAM‐deficient mice did not modify NCAM expression levels in the amygdala or hippocampus, while they showed enhanced anxiety‐like behavior, questioning the involvement of NCAM in this type of behavior. Together, our results strongly support the involvement of NCAM in the amygdala in the consolidation of auditory fear conditioning and highlight increased NCAM expression in the amygdala among the mechanisms whereby stress facilitates fear conditioning processes.     

Capillary electrophoresis of subcellular-sized particles

Utilization of capillary electrophoresis (CE) for characterization and analytical separation of submicron- and micron-sized organic and inorganic particles as well as biological vesicles is reviewed. CE has been applied to charged polystyrene size standards, inorganic and organic colloidal particles, lipoprotein particles, liposomes, microsomes and viruses. These particle separations generally occur in a size-dependent manner and provide values of electrophoretic mobility which are in good agreement with those obtained by other electrophoretic techniques.     

Effect of restricted watering and its combination with root pruning on root growth capacity,water status and food reserves ofPinus caribaea var.hondurensis seedlings

Summary Six-month oldPinus caribaea var.hondurensis seedling were subjected to various regimes of restricted watering and their combinations with root pruning to determine a suitable technique of producing seedlings with optimum physiological condition for successful bare-root planting.From the range of treatments tested, plants subjected to watering once every 9 days (D9) were found to be the hardest in terms of highest root growth capacity, leaf water content, root starch content and in having a moderately low transpiration rate.P. caribaea seedlings subjected to this preconditioning treatment may be more able to survive bare-root planting.     

Adaptations of microbial communities and dissolved organics to seasonal pressures in a mesotrophic coastal Mediterranean lake

In lake ecosystems, changes in eukaryotic and prokaryotic microbes and the concentration and availability of dissolved organic matter (DOM) produced within or supplied to the system by allochthonous sources are components that characterize complex processes in the microbial loop. We address seasonal changes of microbial communities and DOM in the largest Croatian lake, Vrana. This shallow lake is connected to the Adriatic Sea and is impacted by agricultural activity. Microbial community and DOM structure were driven by several environmental stressors, including drought, seawater intrusion and heavy precipitation events. Bacterial composition of different lifestyles (free-living and particle-associated) differed and only a part of the particle-associated bacteria correlated with microbial eukaryotes. Oscillations of cyanobacterial relative abundance along with chlorophyll a revealed a high primary production season characterized by increased levels of autochthonous DOM that promoted bacterial processes of organic matter degradation. From our results, we infer that in coastal freshwater lakes dependent on precipitation-evaporation balance, prolonged dry season coupled with heavy irrigation impact microbial communities at different trophic levels even if salinity increases only slightly and allochthonous DOM inputs decrease. These pressures, if applied more frequently or at higher concentrations, could have the potential to overturn the trophic state of the lake.     

Novel 5-Alkylcarboxamide-2'-Deoxyuridine-5'-Triphosphates for Enzymatic Synthesis of Highly Modified DNA

Russian Journal of Bioorganic Chemistry - The synthesis of deoxyuridine triphosphate derivatives with alkyl groups of various chain length attached to the pyrimidine base by the trans-alkene linker...