HIV Infection, HAART, and Gynecomastia. Epidemiological, Clinical, and Pathogenetic Correlates

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Enzyme-based DNA extraction from zoospores of ruminal fungi

We report here a rapid, efficient and simple method for the extraction of high molecular weight DNA from zoospores of Neocallimastix frontalis EB188. This anaerobic fungus, isolated from bovine digesta, effectively degrades plant fiber in vitro (Barichievich et al. 1990. Appl. Env. Micro. 56:43-48). Our interest in ruminal fungi stems from their ability to degrade wood materials (Joblin et al. 1989. FEMS Micro. Lett. 56:119-122) and their potential use in biomass saccharification. Zoospore DNA synthesis is of particular interest to our laboratory. It is these motile zoospores which colonize and degrade plant materials (Mountfort 1987. FEMS Micro. Rev. 46:501-508). To detail fully this metabolic event, it will be necessary to extract nucleic acids from zoospores. Such procedures have not been reported in the literature. Using acetone drying and enzymatic removal of cell walls, we have isolated high molecular weight DNA from very small amounts of culture. The procedure takes less than one hour and DNA yields are high. The DNA is readily cut with restriction endonucleases and religated efficiently but is otherwise stable. Electrophoretic analysis of the DNA confirmed the presence of repetitive sequences. This procedure will aid the study of DNA replication, DNA repair and DNA RFLP analysis of various strains using small (\u3c1 ml) cultures

Recurring Candida albicans esophagitis in a HIV-infected patient undergoing long-Term antiretroviral therapy, and with absent-negligible immunodeficiency

A patient with HIV infection developed the first episode of AIDS-defining opportunism (severe Candida albicans esophagitis) with an underlying CD4+ lymphocyte count of 1,025 cells/µL. After treatment with a highly active antiretroviral therapy (HAART), taken with insufficient compliance and leaving a residual viral load, our patient suffered from two relapses of esophageal candidiasis, which occurred three months and seven years later, when his CD4+ lymphocyte count was 930 and 439 cells/µL, respectively, and a viral load slightly above 10(4) copies/mL was still present. Also in the HAART era, Candida esophagitis remains one of the most common AIDS-defining diseases, but a presentation with a concurrent CD4+ count above 1,000 cells/µL remains a rare exception, as well as the two isolated, subsequent relapses, occurred with a CD4+ count ranging from 439 to 930 cells/µL, and a residual HIV viremia due to insufficient adherence to the prescribed HAART regimens. Our case report represents the opportunity to revisit the epidemiology and, especially, the pathogenesis of this opportunistic fungal complication in HIV-infected patients and in other subjects at risk, on the ground of an extensive literature review, and to explore possible alternative supporting factors other than the crude absolute CD4+ lymphocyte count, with emphasis on the possible role of a persisting HIV viremia, and other potential contributing factors. Clinicians engaged with immunocompromised patients and subjects with HIV disease, should be aware that a Candida esophagitis may occur and relapse also when the cell-mediated immunity, as measured by a simple CD4+ cell count, do not show relevant abnormalities

Mutagen Killing and Photoreactivation in the Anaerobic Fungus Neocallimastix frontalis EB188

Research was performed to determine if the anaerobic fungus, Neocallimastix frontalis EB 188, a common cellulolytic fungus of ruminants, is capable of photoreactivation and is susceptible to chemical and irradiational mutagenesis. Germination of zoospores and production of colony cellulase was measured after ultraviolet light (UVL), nitrosoguanidine or ethyl methyl sulfonate treatments. This fungus was susceptible to mutagen treatment and capable of photoreactivation after UVL exposure. Such procedures may be useful in the isolation of enhanced cellulase-producing strains

HIV-associated Cutaneous Dissemination of Visceral Leishmaniasis, Despite Negligible Immunodeficiency. Failure of Liposomal Amphotericin B Administration, Followed by Successful Pentamidine-Paromomycin Administration

and post hoc Tukey test. Statistical analysis of survival time was carried out with Kaplan-Meier (Log-Rank) test. Results: The immunized mice with this DNA vaccine presented an important reduction in diameter of lesion and increasing of weight compared to the control mice and was indicated a significant difference between the immunized group and the control groups (p < 0.05). The survival time of the immunized mice was significantly higher than the control groups (p < 0.05) after challenge with Leishmania major. The immunized mice had significantly lower parasite load compared to the control mice (p < 0.05). Conclusion: The findings of this study, indicated that the TSA encoded DNA vaccine induced protection against infection with Leishmania major in mice. In this study, we demonstrated that, the TSA -encoded DNA vaccine may be an excellent candidate for futher vaccine development

Comparative analysis of mesenchymal stromal cells biological properties

The stromal progenitors of mesodermal cells, mesenchymal stromal cells (MSCs), are a heterogeneous population of plastic adherent fibroblast-like cells with extensive proliferative capacity and differentiation potential. Human MSCs have now been isolated from various tissues including bone marrow, muscle, skin, and adipose tissue, the latter being one of the most suitable cell sources for cell therapy, because of its easy accessibility, minimal morbidity, and abundance of cells. Bone marrow and subcutaneous or visceral adipose tissue samples were collected, digested with collagenase if needed, and seeded in Iscove's medium containing 5% human platelet lysate. Nonadherent cells were removed after 2-3 days and the medium was replaced twice a week. Confluent adherent cells were detached, expanded, and analyzed for several biological properties such as morphology, immunophenotype, growth rate, senescence, clonogenicity, differentiation capacity, immunosuppression, and secretion of angiogenic factors. The results show significant differences between lines derived from subcutaneous fat compared to those derived from visceral fat, such as the higher proliferation rate of the first and the strong induction of angiogenesis of the latter. We are convinced that the identification of the peculiarities of MSCs isolated from different tissues will lead to their more accurate use in cell therapy

Constitutive and LPS-stimulated secretome of porcine Vascular Wall-Mesenchymal Stem Cells exerts effects on in vitro endothelial angiogenesis

Background: MSCs secretome is under investigation as an alternative to whole-cell-based therapies, since it is enriched of bioactive molecules: growth factors, cytokines and chemokines. Taking into account the translational value of the pig model, the leading aim of the present paper was to characterize the secretome of porcine Vascular Wall-Mesenchymal Stem Cells (pVW-MSCs) and its change in presence of LPS stimulation. Moreover, considering the importance of angiogenesis in regenerative mechanisms, we analysed the effect of pVW-MSCs secretome on in vitro angiogenesis. Results: Our results demonstrated that conditioned medium from unstimulated pVW-MSCs contained high levels of IL-8, GM-CSF, IFN-\u3b3 and other immunomodulatory proteins: IL-6 IL-18 IL-4 IL-2 IL-10. LPS modulates pVW-MSCs gene expression and secretome composition, in particular a significant increase of IL-6 and IL-8 was observed; conversely, the amount of GM-CSF, IFN-\u3b3, IL-2, IL-4, IL-10 and IL-18 showed a significant transient decrease with the LPS stimulation. Conditioned medium from unstimulated pVW-MSCs induced in vitro endothelial angiogenesis, which is more evident when the conditioned medium was from LPS stimulated pVW-MSCs. Conclusions: The lines of evidence here presented shed a light on possible future application of secretome derived by pVW-MSCs on research studies in translational regenerative medicine

Long-term effect of neonatal inhibition of APP gamma-secretase on hippocampal development in the Ts65Dn mouse model of Down syndrome

Neurogenesis impairment is considered a major determinant of the intellectual disability that characterizes Down syndrome (DS), a genetic condition caused by triplication of chromosome 21. Previous evidence obtained in the Ts65Dn mouse model of DS showed that the triplicated gene APP (amyloid precursor protein) is critically involved in neurogenesis alterations. In particular, excessive levels of AICD (amyloid precursor protein intracellular domain) resulting from APP cleavage by gamma-secretase increase the transcription of Ptch1, a Sonic Hedgehog (Shh) receptor that keeps the mitogenic Shh pathway repressed. Previous evidence showed that neonatal treatment with ELND006, an inhibitor of gamma-secretase, reinstates the Shh pathway and fully restores neurogenesis in Ts65Dn pups. In the framework of potential therapies for DS, it is extremely important to establish whether the positive effects of early intervention are retained after treatment cessation. Therefore, the goal of the current study was to establish whether early treatment with ELND006 leaves an enduring trace in the brain of Ts65Dn mice. Ts65Dn and euploid pups were treated with ELND006 in the postnatal period P3-P15 and the outcome of treatment was examined at ~&nbsp;one month after treatment cessation. We found that in treated Ts65Dn mice the pool of proliferating cells in the hippocampal dentate gyrus (DG) and total number of granule neurons were still restored as was the number of pre- and postsynaptic terminals in the stratum lucidum of CA3, the site of termination of the mossy fibers from the DG. Accordingly, patch-clamp recording from field CA3 showed functional normalization of the input to CA3. Unlike in field CA3, the number of pre- and postsynaptic terminals in the DG of treated Ts65Dn mice was no longer fully restored. The finding that many of the positive effects of neonatal treatment were retained after treatment cessation provides proof of principle demonstration of the efficacy of early inhibition of gamma-secretase for the improvement of brain development in DS