Serological analysis and pathogenic potentials of Wangiella dermatitidis isolated from bats

Methods for the preparation of antigens from clinically isolated cultures of Aspergillus were standardized. Sera from 25 suspected cases of pulmonary aspergillosis were tested against antigens prepared by us, from 4 strains of A. fumigatus and one strain of A. flavus, using the Ouchterlony double diffusion and immunoelectrophoretic techniques.Of the 25 sera tested, 18 reacted positively with antigens of A. fumigatus, one with A. flavus and 2 with both these species. Antigens of two non-pathogenic Aspergilli included in the study failed to react with any of the sera. Our antigen preparations gave more numerous as well as sharper precipitin lines than the commercial Bencard antigens which were used for comparison. Moreover, mycelial antigens from 48 to 96 h old cultures revealed precipitin lines comparable to that of the routine, 4 week old culture filtrate antigens, thus suggesting that the incubation period for obtaining antigens could be cut down considerably.Memoir No. 323 from the Centre of Advanced Study in Botany.Deceased     

Fungi from Geothermal Soils in Yellowstone National Park

Geothermal soils near Amphitheater Springs in Yellowstone National Park were characterized by high temperatures (up to 70°C), high heavy metal content, low pH values (down to pH 2.7), sparse vegetation, and limited organic carbon. From these soils we cultured 16 fungal species. Two of these species were thermophilic, and six were thermotolerant. We cultured only three of these species from nearby cool (0 to 22°C) soils. Transect studies revealed that higher numbers of CFUs occurred in and below the root zone of the perennial plant Dichanthelium lanuginosum (hot springs panic grass). The dynamics of fungal CFUs in geothermal soil and nearby nongeothermal soil were investigated for 12 months by examining soil cores and in situ mesocosms. For all of the fungal species studied, the temperature of the soil from which the organisms were cultured corresponded with their optimum axenic growth temperature.     

Ultrasound imaging in an experimental model of fatty liver disease and cirrhosis in rats

Background

Atypical scrapie was first identified in Norwegian sheep in 1998 and has subsequently been identified in many countries. Retrospective studies have identified cases predating the initial identification of this form of scrapie, and epidemiological studies have indicated that it does not conform to the behaviour of an infectious disease, giving rise to the hypothesis that it represents spontaneous disease. However, atypical scrapie isolates have been shown to be infectious experimentally, through intracerebral inoculation in transgenic mice and sheep. The first successful challenge of a sheep with 'field' atypical scrapie from an homologous donor sheep was reported in 2007.

Results

This study demonstrates that atypical scrapie has distinct clinical, pathological and biochemical characteristics which are maintained on transmission and sub-passage, and which are distinct from other strains of transmissible spongiform encephalopathies in the same host genotype.

Conclusions

Atypical scrapie is consistently transmissible within AHQ homozygous sheep, and the disease phenotype is preserved on sub-passage.     

Anti-IL-2 treatment impairs the expansion of T(reg) cell population during acute malaria and enhances the Th1 cell response at the chronic disease

Plasmodium chabaudi infection induces a rapid and intense splenic CD4(+) T cell response that contributes to both disease pathogenesis and the control of acute parasitemia. The subsequent development of clinical immunity to disease occurs concomitantly with the persistence of low levels of chronic parasitemia. The suppressive activity of regulatory T (T(reg)) cells has been implicated in both development of clinical immunity and parasite persistence. To evaluate whether IL-2 is required to induce and to sustain the suppressive activity of T(reg) cells in malaria, we examined in detail the effects of anti-IL-2 treatment with JES6-1 monoclonal antibody (mAb) on the splenic CD4(+) T cell response during acute and chronic P. chabaudi AS infection in C57BL/6 mice. JES6-1 treatment on days 0, 2 and 4 of infection partially inhibits the expansion of the CD4(+)CD25(+)Foxp3(+) cell population during acute malaria. Despite the concomitant secretion of IL-2 and expression of high affinity IL-2 receptor by large CD4(+) T cells, JES6-1 treatment does not impair effector CD4(+) T cell activation and IFN-γ production. However, at the chronic phase of the disease, an enhancement of cellular and humoral responses occurs in JES6-1-treated mice, with increased production of TNF-α and parasite-specific IgG2a antibodies. Furthermore, JES6-1 mAb completely blocked the in vitro proliferation of CD4(+) T cells from non-treated chronic mice, while it further increased the response of CD4(+) T cells from JES6-1-treated chronic mice. We conclude that JES6-1 treatment impairs the expansion of T(reg) cell population during early P. chabaudi malaria and enhances the Th1 cell response in the late phase of the disease.     

Cytoprotective properties of alpha-tocopherol are related to gene regulation in cultured D-galactosamine-treated human hepatocytes

Vitamin E (alpha-tocopherol) has demonstrated antioxidant activity and gene-regulatory properties. d-Galactosamine (D-GalN)-induced cell death is mediated by nitric oxide in hepatocytes, and it is associated with hepatic steatosis. The beneficial properties of alpha-tocopherol and their relation to oxidative stress and gene regulation were assessed in D-GalN-induced cell death. Hepatocytes were isolated from human liver resections by a collagenase perfusion technique. alpha-Tocopherol (50 microM) was administered at the advanced stages (10 h) of D-GalN-induced cell death in cultured hepatocytes. Cell death, oxidative stress, alpha-tocopherol metabolism, and NF-kappaB-, pregnane X receptor (PXR)-, and peroxisome proliferator-activated receptor (PPAR-alpha)-associated gene regulation were estimated in the hepatocytes. D-GalN increased cell death and alpha-tocopherol metabolism. alpha-Tocopherol exerted a moderate beneficial effect against apoptosis and necrosis induced by D-GalN. Induction (rifampicin) or inhibition (ketoconazole) of alpha-tocopherol metabolism and overexpression of PXR showed that the increase in PXR-related CYP3A4 expression caused by alpha-tocopherol enhanced cell death in hepatocytes. Nevertheless, the reduction in NF-kappaB activation and inducible nitric oxide synthase expression and the enhancement of PPAR-alpha and carnitine palmitoyl transferase gene expression by alpha-tocopherol may be relevant for cell survival. In conclusion, the cytoprotective properties of alpha-tocopherol are mostly related to gene regulation rather than to antioxidant activity in toxin-induced cell death in hepatocytes.     

A disappearing oasis in the semi-arid Chaco: Deficient palm regeneration and establishment

Palm forests of Copernicia alba are a rare habitat in the semi-arid Chaco of Northwestern Argentina, are centres of high species diversity, and provide key resources for many species. Our goal was to assess the conservation status of five C. alba patches in Northwestern Argentina: Reserve; Embarcación; Palma Sola; Vinalito; and, Talar. We compared patches to identify the sites with greatest conservation needs based on four criteria: population size structure; palm density (of individuals with height >1.30 m); probability of seedlings being browsed; and, presence of potential seed dispersers. We found that three (Embarcación, Reserve, and Talar) out of five sites had palm densities greater than 200 individuals/ha and only one site (Embarcación) showed a reverse J-shaped size structure for height. Reserve and Embarcación had the greatest probability of seedlings being browsed (0.99 ± 0.01 and 0.88 ± 0.12, respectively). A total of 14 potential disperser species of mammals and birds were recorded across the five sites. Only Reserve harboured all of the potential dispersers, but at least two potential disperser species were recorded at the other sites. None of the palm patches studied had an adequate conservation status. However, Embarcación met three out of four criteria, and therefore it can be considered to have the best conservation status in the semi-arid Chaco. Palma Sola and Vinalito have the greatest conservation needs. To conserve C. alba in Northwestern Argentina, strategies are needed that ensure seedling establishment for future populations to reach an adequate density and structure.     

Heterogeneity of the digestible insoluble fiber of selected forages in situ

Long-term in situ incubations were performed to verify the likelihood of the heterogeneity concept of the potentially digestible fraction of the insoluble fiber (NDFom) by fitting both heterogeneous and homogeneous potentially digestible NDFom versions of a generalized compartmental model of digestion (GCMD). Corn silage and eleven tropical grasses and alfalfa hay were studied. Data were gathered from a study in which forage samples in nylon bags were incubated in rumen cannulated steers so that three profiles per forage were generated. The incubation endpoint was used to form sets of time profiles. The original set consisted of profiles ending at 1440 h, and the other two were formed by using 96 and 240 h as the incubation endpoints, respectively. The indigestible residue was estimated using nonlinear least squares or by assuming it to be 2.4 times lignin determined by the sulphuric acid method (Lignin (sa)). Therefore, eight different models were evaluated by combining end points of digestion, and the homogeneous and heterogeneous versions of GCMD with the two ways of estimating the indigestible residue. The likelihood of the models was assessed by computing Akaike information criteria. The effects of forage, model, and their interaction were analyzed by taking model as a repeated measurement. Heterogeneity of the potentially degradable fraction for NDFom was detected with long-term incubation trials (up to 1440 h) for some forages, and the introduction of the 2.4×Lignin (sa) as a direct measure of the indigestible residue improved the likelihood of the heterogeneous version of GCMD. The forage by model interaction was significant for many comparable parameter estimates, which means that specific and inconsistent results for models within forages were produced depending on the definition of the incubation end-point. The indigestible residue was overestimated with short-term incubation profiles, but the overestimation was lower for the profiles ending at 240 h whether compared to profiles ending at 96 h. Given the likelihood of the heterogeneous version of GCMD fitted to profiles ending at 1440 h and at 240 h for some forages, the heterogeneity concept should be investigated whenever the research interest relies on estimating the kinetic attributes of the degradation profiles of the NDFom in situ.     

In situ quantification of aberrant p53 in colorectal neoplasia

Aberrant p53 protein accumulation was measured immunohistologically in 342 colorectal paraffin-embedded tissue sections from 115 patients (24 with adenocarcinoma, 59 with adenoma and 32 'hospital controls'). Subjective scoring was compared with quantitative cell imaging, including dichotomous (p53⁺/p53^-) status, ng p53^mut mg^-1 enterocyte protein, and tumour burden and patient body 'burden' of aberrant p53. A total of 62.5% cancer patients, 23.7% adenoma patients and 3.1% hospital controls were accorded p53⁺ status on the basis of p53 quantification. Quantitative p53⁺/p53^- assignment had a stronger inverse association with survival (χ²=6.17, p=0.013, Kaplan-Meier test) than subjective 'visual estimation' (χ²=0.57, p=0.449). There was a strong inverse relationship between the p53 'body burden' and the months of post-diagnosis survival (hazard ratio=1.42, p=0.0004, Cox proportional hazards). Absolute quantification for inactivated p53 permits objective and reproducible scoring, adjusts for intra-laboratory immunostaining 'batch effects', corrects for fixation artefacts, and standardizes for inter-laboratory differences in fixation, antibody selection and staining method. Clinically, in situ quantification of p53 will permit more accurate survival prognoses and will inform therapy selection and dose. Ultimately, accurate quantitative tissue/blood p53 correlations may provide a minimally invasive and systemic surrogate measure for these same clinical purposes.     

Risk Factors for Death from Influenza A(H1N1)pdm09, State of S?o Paulo,Brazil, 2009

This case-control study aimed to assess the risk factors for death from influenza A(H1N1)pdm09 in patients with laboratory confirmation, who had severe acute respiratory illness-SARI and were hospitalized between June 28^th and August 29^th 2009, in the metropolitan regions of São Paulo and Campinas, Brazil. Medical charts of all the 193 patients who died (cases) and the 386 randomly selected patients who recovered (controls) were investigated in 177 hospitals. Household interviews were conducted with those who had survived and the closest relative of those who had died. 73.6% of cases and 38.1% of controls were at risk of developing influenza-related complications. The 18-to-59-year age group (OR = 2.31, 95%CI: 1.31–4.10 (reference up to 18 years of age)), presence of risk conditions for severity of influenza (OR = 1.99, 95%CI: 1.11–3.57, if one or OR = 6.05, 95%CI: 2.76–13.28, if more than one), obesity (OR = 2.73, 95%CI: 1.28–5.83), immunosuppression (OR = 3.43, 95%CI: 1.28–9.19), and search for previous care associated with the hospitalization (OR = 3.35, 95%CI: 1.75–6.40) were risk factors for death. Antiviral treatment performed within 72 hours of the onset of symptoms (OR = 0.17, 95%CI: 0.08–0.37, if within 48hours, and OR = 0.30, 95%CI: 0.11–0.81, if between 48 and 72 hours) was protective against death. The identification of high-risk patients and early treatment are important factors for reducing morbi-mortality from influenza.