Tuberculosis infection in HIV vs. non-HIV patients.

OBJECTIVES: Tuberculosis (TB) is the most common opportunistic infection and cause of mortality among people living with HIV, and it is possible that it may also influence the evolution of the HIV infection. We assessed the differences between HIV-positive and -negative people infected with TB. METHODS: The present study is a cross-sectional retrospective study by electronic record revision. We included patients admitted to a tertiary hospital with a diagnosis of TB between 2011 and 2016, comparing those with HIV coinfection with non-HIV patients, according to demographic and clinical characteristics. RESULTS: This study included 591 patients, of whom 32% were HIV-coinfected. HIV-TB patients were younger, with a predominance of male gender. Considering TB risk factors, there was a higher prevalence of homelessness and intravenous drug use in the HIV group. In the non-HIV group, direct contact with other patients with TB and immunosuppression were more prevalent. Relative to TB characteristics, the HIV-coinfected group presents with a higher prevalence of disseminated disease and a higher occurrence of previous TB infection. Cancer was the most frequent cause of immunosuppression in the HIV group and the number testing positive for TB via microbiological culture was lower. Assessment of microbiological resistance and in-hospital mortality showed similar numbers in both groups. CONCLUSIONS: There are few papers comparing clinical course of TB between HIV-infected and non-infected patients. Our study differs from others in the literature as we focused on a country with middling incidence of TB and further characterized the differences between HIV-infected and non-infected patients which can contribute to the management of these patients.

Pneumocystosis pneumonia: A comparison study between HIV and non-HIV immunocompromised patients.

Pneumocystis pneumonia (PCP) is caused by the fungus Pneumocystis jirovecii, and its incidence has been on the rise in immunosuppressed patients without HIV. We performed a cross sectional study in patients with PCP and assessed demographic, clinical presentation and outcome measures such as mechanical ventilation and mortality differences between HIV and non-HIV patients. The two groups were statistically significantly different, with the HIV group being younger (45.5 years vs 55.9 years, p-value 0.001) and mostly composed of male patients (69% vs 31%, p-value <0.001). Also, the HIV patients had higher percentage of respiratory complaints (90% vs 68%, p-value 0.02) and lactate dehydrogenase elevation (73% vs 40%, p-value 0.001). In contrast, non-HIV patients had worse outcomes with higher incidence of invasive mechanical ventilation (23% vs 46%, p-value 0.005) and in-hospital mortality (13% vs 37%, p-value 0.002). These results reflect the literature and should raise awareness to a potentially fatal medical situation of increasing incidence.

Strength and Aerobic Physical Exercises Are Able to Increase Survival of Toxoplasma gondii-Infected C57BL/6 Mice by Interfering in the IFN-γ Expression.

Physical exercise has been implicated in several immunophysiological improvements, particularly during the aging process, when an immunocompromised status could be established. Toxoplasma gondii is a protozoan parasite that causes a widespread opportunistic infection, which may present severe consequences, mainly to the fetus and immunocompromised patients. It is estimated that one-third of the human population worldwide has been infected by this parasite, being the reactivation during immunesenescence an unexplored public health issue. The major purpose of the present study was to observe the immunophysiological differences between exercised vs. sedentary C57BL/6 male mice that have been experimentally infected by T. gondii. In the first set of experiments, the animals were infected after exercising and three groups were set up: experimental groups-infected sedentary (IS, n = 6); infected exercised (IEx, n = 6) and control group-non-infected sedentary (NIS, n = 6). When stimulated in vitro by T. gondii-soluble tachyzoite antigen, it was found that splenocytes from exercised group produced higher levels of IFN-γ, as well as of IFN-γ/IL-10 ratios in comparison with splenocytes from sedentary animals (P < 0.001). However, it was not found significant differences concerning quantification of T. gondii genomic DNA by qRT-PCR and immunohistochemistry analysis in brain cysts from both group of animals (P > 0.05). In order to further investigate the consequences of these data for the host, a second set of experiments was performed, when the animals were infected before exercising and four groups of animals were established for comparison purpose, as follows: experimental groups-infected sedentary (IS, n = 7); infected exercised (IEx, n = 6) and control groups-non-infected sedentary (NIS, n = 6) and non-infected exercised (NIEx, n = 6). It was found significant differences in the survival rates of the exercised group the animals, as they survived longer than sedentary groups (P = 0.0005). In both sets of experiments, mice have been submitted to moderate exercises: aerobic (14 m/min; 3 x/week) and strength (60-80% of one maximum repetition; 2 x/week). Overall, our findings are showing that the aerobic and strength exercises are able to modulate immune response against T. gondii infection, being these immunological features beneficial to the host.

An innovative challenge test for solid cosmetics using freeze-dried microorganisms and electrical methods.

Freeze-dried bacteria and fungi were used as inoculum in 28 days' PET. An electrical method was used in replacement of the conventional plate count method. The use of freeze-dried microorganisms in association with the electrical method can minimize the workload and the variability involved in PET for cosmetic powders.

Fluorescent ester dye-based assays for the in vitro measurement of Neospora caninum proliferation.

Techniques for the measurement of parasite loads in different experimental models have evolved throughout the years. The quantification of stained slides using regular cytological stains is currently the most common technique. However, this modality of evaluation is labor-intensive, and the interpretation of the results is subjective because the successes of the assays mainly rely on the abilities of the professionals involved. Moreover, the novel genetic manipulation techniques that are commonly applied for closely related Toxoplasma gondii have not yet been developed for Neospora caninum. Thus, we aimed to develop a simple protocol for parasite quantification using pre-stained N. caninum tachyzoites and fluorescent probes based on ester compounds (i.e., CFSE and DDAO). For this purpose, we employed a quantification procedure based on flow cytometry analysis. Pre-stained parasites were also examined with a fluorescent microscope, which revealed that both dyes were detectable. Direct comparison of the numbers of CFSE+ and DDAO+ cells to the values obtained with classical cytology techniques yielded statistically comparable results that also accorded with genomic DNA amplification results. Although the fluorescence emitted by DDAO was more intense and provided better discrimination between the populations of parasitized cells, CFSE+ tachyzoites were detected for several days. In conclusion, this study describes a simple, fast, low-cost and reproducible protocol for N. caninum quantification that is based on parasite pre-staining with fluorescent ester-based probes.

Adolescent social isolation enhances the plasmalemmal density of NMDA NR1 subunits in dendritic spines of principal neurons in the basolateral amygdala of adult mice.

Social isolation during the vulnerable period of adolescence produces emotional dysregulation manifested by abnormalities in adult behaviors that require emotional processing. The affected brain regions may include the basolateral amygdala (BLA), where plasticity of glutamatergic synapses in principal neurons plays a role in conditioned emotional responses. This plasticity is dependent on NMDA receptor trafficking denoted by intracellular mobilization of the obligatory NR1 NMDA subunit. We tested the hypothesis that the psychosocial stress of adolescent social isolation (ASI) produces a lasting change in NMDA receptor distribution in principal neurons in the BLA of adults that express maladaptive emotional responses to sensory cues. For this, we used behavioral testing and dual electron microscopic immunolabeling of NR1 and calcium calmodulin-dependent protein kinase II (CaMKII), a protein predominantly expressed in principal neurons of the BLA in adult C57Bl/6 mice housed in isolation or in social groups from post-weaning day 22 until adulthood (∼3 months of age). The isolates showed persistent deficits in sensorimotor gating evidenced by altered prepulse inhibition (PPI) of acoustic startle and hyperlocomotor activity in a novel environment. Immunogold-silver labeling for NR1 alone or together with CaMKII was seen in many somatodendritic profiles in the BLA of all mice irrespective of rearing conditions. However, isolates compared with group-reared mice had a significantly lower cytoplasmic (4.72 ± 0.517 vs 6.31 ± 0.517) and higher plasmalemmal (0.397 ± 0.0779 vs 0.216 ± 0.026) density of NR1 immunogold particles in CaMKII-containing dendritic spines. There was no rearing-dependent difference in the size or number of these spines or those of other dendritic profiles within the neuropil, which also failed to show an impact of ASI on NR1 immunogold labeling. These results provide the first evidence that ASI enhances the surface trafficking of NMDA receptors in dendritic spines of principal neurons in the BLA of adult mice showing maladaptive behaviors that are consistent with emotional dysregulation.

New development on materials and techniques used in the heraldic designs of illuminated Manueline foral charters by multi-analytical methods.

In the illuminated foral charters commissioned by D. Manuel I, King of Portugal (1495-1521), heraldry played a prominent role. In this work, royal emblems corresponding to the third heraldic design, applied to the Charter of Couto de Teixedo, are studied by means of EDXRF, SEM-EDS, micro-Raman, micro-FTIR and HPLC-DAD to characterize the materials and techniques used to produce this masterpiece of Portuguese Renaissance. By comparing this data with the results previously obtained in other three charters, this research allows for the first time an accurate insight into the reformed manuscripts production, especially since the illuminated charters under consideration were all investigated on the basis of similar micro-spectroscopic techniques. Inks, metal coatings, pigments, organic dyes, fillers or extenders, and binding media are discussed, and their use enlightened by the politic and legal messages the foral charters had to convey. Parchment analysis allowed us to identify the animal species, and all undertaken processes along its preparation. HPLC-DAD allowed us to differentiate the gums used. CaSO4 was observed in the parchment as a degradation product of CaCO3 used in parchment production in the presence of metal sulfates coming from the ink. Pigments used in the different colors as well as some specific phases for some colors were identified. Silvering and gilding processes, metal caratage and silver oxidation compounds were also objects of the present study.

Multi-molecular markers and metals as tracers of organic matter inputs and contamination status from an Environmental Protection Area in the SW Atlantic (Laranjeiras Bay, Brazil).

The sources and concentrations of aliphatic hydrocarbons (AHs) and polycyclic aromatic hydrocarbons (PAHs), faecal and biogenic sterols, and trace metals at 10 sampling sites located in Laranjeiras Bay, a large Environmental Protection Area in the southern Atlantic region of Brazil, were determined to assess the sources of organic matter and the contamination status of estuarine sediments. Organic compounds were determined by GC-FID and GC-MS, and ICP-OES was used to evaluate trace metals. The total AHs concentration ranged from 0.28 to 8.19 µg g(-1), and n-C(29) and n-C(31) alkanes were predominant, indicating significant inputs from higher terrestrial plants. Unresolved complex mixtures (UCM) were not detected at any site, suggesting that the study area was not significantly contaminated by fossil fuels. The total PAH concentration varied from 3.85 to 89.2 ng g(-1). The ratio between selected PAH isomers showed that combustion of biomass, coal, and petroleum is the main source of PAHs in the study area. The concentrations of the faecal sterols coprostanol and epicoprostanol were below the detection limits, suggesting that sewage was not a significant contributor to sedimentary organic matter. The concentrations of the trace metals (As, Cr, Cu, Ni, Pb and Zn) were low, except near sites located at the mouths of rivers that discharge into the study area and near urbanised regions (Paranaguá city and the adjoining harbour). In general, the concentrations of PAHs were below the threshold effect concentrations (TEL) levels. Although the As, Cr and Ni concentrations were above the TEL levels, the study area can be considered as preserved from human activities.

Avaliação de indicadores biológicos na validação de processos de esterilização de isoladores por peróxidos de hidrogênio / Assessment of biological indicators in the validation of isolator decontamination with hydrogen peroxide

Os isoladores permitem a aplicação de descontaminação por gases, resultando em ambiente estéril. Esta característica, adicionada a possibilidade de não interferência humana no processo, torna o emprego dos isoladores consideravelmente vantajosa quando comparada com a performance dos processos em salas limpas convencionais. A descontaminação empregando peróxido de hidrogênio é vantajosa em relação a outros métodos disponíveis uma vez que é de fácil remoção após aplicação; sendo água e oxigênio seus produtos de degradação, apresenta boa compatibilidade com materiais usualmente empregados nas áreas produtivas; e seu custo é relativamente baixo. O propósito deste estudo foi demonstrar que o Geobacillus stearothermophilus (ATCC 12980) é o microrganismo mais resistente quando comparado com microrganismos isolados presentes em áreas produtivas, assim como avaliar o melhor material a servir de suporte durante a validação do processo de descontaminação das superfícies internas do isolador e externas de materiais presentes. Bacillus sp., Micrococcus luteus, Corynebacterium sp., Staphylococcus sp. e Penicilium sp. foram os microrganismos que apresentaram maior incidência na área produtiva. Aço inoxidável é o material mais adequado a ser usado como suporte para o emprego do peróxido de hidrogênio, por ser inerte e o principal componente dos isoladores e por não demonstrar incompatibilidade com o agente esterilizante. Os resultados obtidos nesta etapa do estudo demonstraram que o Geobacillus stearothermophilus é o microrganismo mais resistente para ser utilizado na avaliação da eficácia do peróxido de hidrogênio quando comparado com aqueles microrganismos encontrados na flora normal. Adicionalmente, o melhor suporte é o aço inoxidável, significando que os indicadores biológicos comercialmente disponíveis neste material são a melhor opção para este propósito.

Isolators allow decontamination gases to be employed to create a sterile processing environment. This feature, added to the potential removal of human interference in the process, makes the use of isolators rather advantageous, compared to performing aseptic processes in conventional clean rooms. Decontamination with vaporized hydrogen peroxide (VHP) offers several advantages over other available methods, as it decomposes to water and oxygen and is thus easy to remove after use, is highly compatible with materials usually employed in production areas and it is relatively cheap. The aims of this study were to prove that Geobacillus stearothermophilus (ATCC 12980) is more resistant than microorganisms isolated from the normal production area flora and to determine the best material to serve as a support during validation of the decontamination of the inner surfaces of isolators and outer surfaces of materials inside them. Bacillus sp., Micrococcus luteus, Corynebacterium, Staphylococcus sp. and Penicillium sp. were the microorganisms of highest incidence among those identified in the production area. Stainless steel is the best material to be used as a support for the VHP treatment of specimens, as it is inert and the main component of isolators and showed no incompatibility with this sterilizing agent. The results obtained in this phase of the experiment proved that Geobacillus stearothermophilus is the most resistant microorganism with which to challenge the effectiveness of hydrogen peroxide, when tested against species of the normal flora. Secondly, the best support material is stainless steel, showing that the commercial bioindicators available on the market with this support material are scientifically proved to be the best choice for this purpose.

The efficacy and stability of five sanitizing agents challenged with reference microorganisms and clean area isolates / Eficácia e estabilidade de cinco agentes sanitizantes avaliados frente a microrganismos de referência e isolados de áreas limpas

The antimicrobial activity of five sanitizing agents employed in clean areas designated for the pharmaceutical manufacturing of sterile products was tested against nine microorganisms, including four microorganisms from the clean area microbiota. The method consisted of challenging 5 mL of each sanitizing agent - 70% isopropyl alcohol, 0.4% LPH®, 1.16% hydrogen peroxide, 4% hydrogen peroxide, 1% Bioper® and 5% phenol - with 0.1mL each of concentrated suspensions (105 ? 106 CFU/mL) of Staphylococcus aureus, Candida albicans, Corynebacterium sp., Micrococcus luteus, Escherichia coli, Aspergillus niger, Bacillus subtilis, Staphylococcus sp. and Bacillus sp. for 10 minutes, followed by serial dilutions and plating. The results demonstrated that the five agents were effective against S. aureus, C. albicans, Corynebacterium sp., and M. luteus. The same was true of E. coli, except that isopropyl alcohol showed low levels of inactivation. With A. niger, isopropyl alcohol, 0.4% LPH® and hydrogen peroxide were more effective and 5% phenol and 1% Bioper® less effective. 1% Bioper® and 4% hydrogen peroxide showed greater inactivation of Staphylococcus sp., Bacillus sp. and B. subtilis than the other agents. Against S. aureus, C. albicans, Corynebacterium sp. and M. luteus, 5% phenol showed similar activity to other agents, while with A. niger, B. subtilis, Staphylococcus sp. and Bacillus sp., it was similar to or less active than the other agents. It was demonstrated that two microorganisms from the clean area microbiota, Staphylococcus sp. and Bacillus sp., were the most difficult to eradicate, requiring more frequent application of hydrogen peroxide and 1% Bioper® than the other strains.

O objetivo deste estudo é avaliar a atividade antimicrobiana de cinco agentes sanitizantes empregados em áreas limpas construídas para a fabricação de produtos farmacêuticos estéreis contra nove microrganismos, incluindo quatro microrganismos oriundos da área limpa. A metodologia constituiu em desafiar 5 mL de cada agente sanitizante, álcool isopropílico 70%, LPH® 0,400%, peróxido de hidrogênio 1,160% e 4%, Bioper® 1% e fenol 5% com 0,1 mL de suspensão concentrada (105 ? 106 UFC/mL) de Staphylococcus aureus, Candida albicans, Corynebacterium sp., Micrococcus luteus, Escherichia coli, Aspergillus niger, Bacillus subtilis, Staphylococcus sp. e Bacillus sp. individualmente por 10 minutos, seguido de diluições seriadas e plaqueamento. Os resultados demonstraram que os cinco agentes sanitizantes foram efetivos contra S. aureus, C. albicans, Corynebacterium sp., e M. luteus. Os mesmos resultados foram observados com E. coli, exceto para o álcool isopropílico, que demonstrou baixos níveis de inativação. Contra A. niger, álcool isopropílico, 0.4% LPH® e peróxido de hidrogênio foram mais efetivos e fenol e Bioper® menos efetivos. Bioper® e peróxido de hidrogênio 4% demonstraram altos níveis de inativação de Staphylococcus sp., Bacillus sp. e B. subtilis quando comparados com outros agentes. Fenol demonstrou atividade antimicrobiana similar aos outros agentes contra S. aureus, C. albicans, Corynebacterium sp. e M. luteus. Contra A. niger, B. subtilis, Staphylococcus sp. e Bacillus sp., a atividade antimicrobiana do fenol foi similar ou inferior a dos outros agentes. Foi demonstrado que os microrganismos isolados da área limpa, Staphylococcus sp. e Bacillus sp., foram os que apresentaram maior dificuldade para inativar, sendo necessária a aplicação de peróxido de hidrogênio e Bioper® , com maior frequência.

Análise da carga microbiana de implantes mamários de silicone / Microbial load analysis in silicone gel breast implants

Silicone breast implants consist of biomaterials widely used in breast reconstitution surgeries or in mammary augmentation for esthetic reasons. A preliminary stage of the implant production process is vulcanization, which consists of heating the implant to 165±5ºC for approximately 9 hours. The aim of this work was to evaluate the bioburden of silicone breast implants prior to the vulcanization process and the decline in bioburden due to this process, and to confirm the sterility of the gel contained in the membrane. Breast implant production stages were evaluated by microbial counting in different steps, according to the USP 32 methodology. To evaluation of decrease in microbial load, spores strips were introduced inside the implant, and after vulcanization cycles the strips were removed from the implant. The strips were transferred to tubes containing TSB, followed by incubation for 7 days at 30-35ºC. The results obtained showed that the level of microbial contamination of gel implants is relatively low, and that vulcanization allowed for the inactivation of up to 108 spores. This study led us to the conclusion that vulcanization leaded to sterility of the gel inside the product. Thus, the final sterilizing process contributed to an increase in the Sterility Assurance Level.1.

Os implantes mamários de silicone constituem-se em biomateriais que têm sido amplamente utilizados em cirurgias para reconstituição da mama ou para o aumento do tamanho da mama por motivos estéticos. Uma etapa preliminar do processo produtivo do implante é a vulcanização, que consiste no aquecimento do implante a 165±5ºC por aproximadamente 9 horas. O objetivo deste estudo foi avaliar a carga microbiana dos implantes mamários de silicone antes do processo de vulcanização, o decaimento da carga microbiana neste processo e confirmar a esterilidade do gel contidointernamente à membrana. Os estágios do processo produtivo dos implantes mamários foram avaliados pela contagem microbiana em diferentes etapas, de acordo com a metodologia da USP 32. Para avaliação do decaimento da carga microbiana, tiras de esporos foram introduzidas no interior do implante e após os ciclos de vulcanização foram retiradas do implante. As tiras foram transferidas para tubos contendo TSB, seguidos pela incubação por 7 dias a 30-35ºC. Os resultados obtidos mostraram que o nível de contaminação microbiana dos implantes gelatinosos é relativamente baixo e que a vulcanização possibilitou a inativação de até 108 esporos. Este estudo nos leva à conclusão que a vulcanização levou à esterilidade do gel interno ao produto. Desta forma, o processo esterilizante final contribuiu para um aumento no Nível de Garantia de Esterilidade.1.

Validação de método de doseamento e aplicação em estudo de equivalência farmacêutica de solução injetável de metronidazol / Validation of metronidazole assay methods and application to study of pharmaceutical equivalence of injectable solutions of this drug

O presente trabalho tem por objetivos validar métodos por espectrofotometria de absorção no ultravioleta e cromatografia líquida de alta eficiência para o doseamento de metronidazol em solução injetável e aplicá-los em estudo de equivalência farmacêutica entre medicamento de referência, genérico e similar. Os métodos propostos para doseamento de metronidazol em solução injetável por espectrofotometria de absorção no ultravioleta e por cromatografia líquida de alta eficiência foram validados, mostrando especificidade / seletividade, linearidade e faixa linear, limite de detecção /quantificação, exatidão e precisão adequados para o uso pretendido. Os medicamentos foram avaliados quanto aos testes de determinação de pH, volume médio, identificação por espectrofotometria de absorção no infravermelho, identificação por cromatografia líquida de alta eficiência, esterilidade, endotoxinas bacterianas e doseamento por espectrofotometria de absorção no ultravioleta e por cromatografia líquida de alta eficiência. Os três medicamentos atenderam as especificações para os testes avaliados e, portanto, podem ser considerados apresentando equivalência farmacêutica.

The aim of this study was to validate analytical methods, based on UV absorption spectrophotometry and high performance liquid chromatography (HPLC), to assay metronidazole supplied in injectable solutions, and to employ these methods in a study of the pharmaceutical equivalence of the original brand name medicine (?reference?), generic and similar (brand) medicines. The methods proposed for the metronidazole assay were validated, showing adequate specificity/selectivity, linearity, linear range, detection and quantitation limits, accuracy and precision for the intended purpose. The injectable solutions were then tested to determine pH, volume and identity (by IR spectrophotometry), to verify sterility, detect bacterial endotoxins and assay the drug by the proposed UV spectrophotometric and HPLC methods. All three medicines met the requirements in all tests performed and, therefore, can be considered pharmaceutically equivalent.

Validação de método de doseamento para aciclovir eaplicação em estudo de equivalência farmacêutica decreme contendo aciclovir / Validation of acyclovir assay method and its use toverify pharmaceutical equivalence of creamscontaining acyclovir

O presente trabalho teve por objetivos validar método para o doseamento de aciclovir em creme por espectrofotometria de absorção no ultravioleta e aplicá-lo em estudo de equivalência farmacêutica entre medicamento de referência, genérico e similar. O método proposto para doseamento de aciclovir em creme por espectrofotometria de absorção no ultravioleta foi validado, mostrando especificidade/ seletividade, linearidade e faixa linear, limite de detecção /quantificação, exatidão e precisão adequados para o uso pretendido. Os medicamentos foram avaliados quanto aos testes de peso médio, limite de guanina por cromatografia em camada delgada, identificação por espectrofotometria de absorção no ultravioleta, contagem microbiana de bactérias e fungos, pesquisa de microrganismos patógenos e doseamento por espectrofotometria de absorção no ultravioleta. Os três medicamentos atenderam as especificações para os testes avaliados e, portanto, podem ser considerados equivalentes farmacêuticos.

The aim of this study was to validate a UV absorption spectrophotometric method to assay acyclovir in cream and to use it to verify the pharmaceutical equivalence of the original brand-name, generic and similar (brand) medicines. The method proposed for acyclovir cream was validated, showing adequate specificity/selectivity, linearity and linear range, detection and quantitation limits, accuracy and precision. The medicines were tested for average weight, guanine contents within limits (by thin-layer chromatography), drug identity (by UV spectrophotometry), bacterial and fungal counts and presence of pathogens, and were assayed by UV spectrophotometry. All medicines met the requirements in all these tests and can, therefore, be considered equivalent.

Detection of all four human metapneumovirus subtypes in nasopharyngeal specimens from children with respiratory disease in Uberlândia, Brazil.

The human metapneumovirus (hMPV) is a pathogen of the respiratory tract identified first in the Netherlands in 2001 and since then it has been detected worldwide. The purpose of this study was to identify and characterize hMPV in samples collected from children <5 years presenting with acute respiratory disease (ARD) seen at a public hospital in Uberlândia, in Southeastern Brazil. One hundred fourteen nasopharyngeal aspirates (NPAs) samples that were negative for the presence of nine other respiratory viruses were tested by reverse transcription polymerase chain reaction (RT-PCR) for the presence of hMPV RNA. Fourteen out of 114 (12.3%) samples were positive for presence of hMPV RNA. PCR products, obtained by the amplification of partial nucleotide sequence of gene N, were sequenced and compared with sequences deposited in GenBank. Sequences from eight samples were obtained and all four subtypes were identified. Also, the recently proposed sublineages "a" and "b" of subtype A2 were found; mean age was 21 months old; upper respiratory tract infection (URTI) was the most common clinical symptom; the virus was detected in samples collected from March to November, a period that corresponds to late summer to mid-spring in Brazil. This is the first study to describe the circulation of all hMPV subtypes in Minas Gerais state.

Netrin-1 interacts with amyloid precursor protein and regulates amyloid-beta production.

The beta-amyloid precursor protein (APP) is an orphan transmembrane receptor whose physiological role is largely unknown. APP is cleaved by proteases generating amyloid-beta (Abeta) peptide, the main component of the amyloid plaques that are associated with Alzheimer's disease. Here, we show that APP binds netrin-1, a multifunctional guidance and trophic factor. Netrin-1 binding modulates APP signaling triggering APP intracellular domain (AICD)-dependent gene transcription. Furthermore, netrin-1 binding suppresses Abeta peptide production in brain slices from Alzheimer model transgenic mice. In this mouse model, decreased netrin-1 expression is associated with increased Abeta concentration, thus supporting netrin-1 as a key regulator of Abeta production. Finally, we show that netrin-1 brain administration in Alzheimer model transgenic mice may be associated with an amelioration of the Alzheimer's phenotype.

Protective effects of beta-glucan extracted from barley against benzo[a]pyrene-induced DNA damage in hepatic cell HepG2.

The aim of the present study was to assess the genotoxic and antigenotoxic effect of beta-glucan (BG) extracted from barley. The genotoxicity of BG was tested in the single-cell gel electrophoresis assays (SCGE)/HepG2 test system. Moreover, the protective effects of BG against the genotoxicity of B[a]P were studied to delineate its mechanism of antigenotoxicity using four different treatment protocols - pre-treatment, simultaneous simple, simultaneous with pre-incubation, and post-treatment. The results showed that the compound itself was devoid of mutagenic activity at the three lower concentrations studied (1, 5, and 25microg/mL); however, genotoxic and cytotoxic effects were seen at 100 and 200microg/mL, respectively. In combination experiments with B[a]P, pronounced inhibition of DNA migration in the SCGE assay was observed in the two simultaneous treatments, and a smaller reduction was observed in the two other treatments. Thus, the data suggest that BG acts through binding to the genotoxic compound or capturing free radicals produced during its activation. However, the protective effects observed with pre-treatment and post-treatment suggest that the BG may be modulating cell metabolism.

Contemporary analysis of the influence of acute kidney injury after reduced intensity conditioning haematopoietic cell transplantation on long-term survival.

We evaluated retrospectively the incidence of acute kidney injury (AKI), defined by risk, injury, failure, loss and end-stage kidney disease (RIFLE) and its influence on long-term survival, in 82 patients aged 18-60 years who underwent a reduced intensity conditioning (RIC) haematopoietic cell transplantation (HCT). Patients (53.6%) developed AKI after HCT: 25% were on risk, 45.5% on injury and 29.5% on failure. In all, 64 patients survived after 100 days of post transplant and were available for long-term survival analysis. At follow-up, 43.7% of patients died. A 5-year overall survival of AKI patients was 41.6% as compared with 67.1% for those who did not develop AKI (P=0.028), and decreased according to AKI severity (risk, 55.6%; injury plus failure, 33.3%; P=0.045). After adjusting for age, history of cardiovascular disease, high-risk disease and chronic GVHD, AKI predicted 5-year overall mortality (AKI: adjusted hazards ratio (AHR), 2.36, 95% CI: 1.03-5.37; P=0.041). Moreover, moderate and severe AKI (injury plus failure) was also associated with an increased 5-year overall mortality (injury plus failure: AHR, 1.64, 95% CI: 1.06-2.54; P=0.024). According to RIFLE, 53.6% of patients had AKI after RIC HCT. Such patients have poor long-term survival, particularly in moderate or severe AKI.

Activated status of basophils in chronic urticaria leads to interleukin-3 hyper-responsiveness and enhancement of histamine release induced by anti-IgE stimulus.

BACKGROUND: Basophils and mast cells are the main target cells in chronic idiopathic urticaria (CIU). Besides the basopenia, intrinsic defects of the anti-IgE cross-linking signalling pathway of basophils have been described in CIU. OBJECTIVES: We sought to investigate the profile of expression of activation markers on basophils of patients with CIU and to explore the effect of interleukin (IL)-3 priming upon anti-IgE cross-linking stimuli through expression of activation markers and basophil histamine releasability. METHODS: Evaluation of the surface expression of FcepsilonRIalpha, CD63, CD203c and CD123 on whole blood basophils of patients with CIU undergoing autologous serum skin test (ASST) was performed by flow cytometry. The effect of pretreatment with IL-3 in the anti-IgE response was analysed by the expression of basophil activation markers and histamine release using enzyme-linked immunosorbent assay. RESULTS: Blood basophils of patients with CIU were reduced in number and displayed increased surface expression of FcepsilonRIalpha, which was positively correlated with the IgE serum levels. Upregulation of expression of both surface markers CD203c and CD63 was verified on basophils of patients with CIU, regardless of ASST response. High expression of IL-3 receptor on basophils was detected only in ASST+ patients with CIU. Pretreatment with IL-3 upregulated CD203c expression concomitantly with the excreting function of blood basophils and induced a quick hyper-responsiveness to anti-IgE cross-linking on basophils of patients with CIU compared with healthy controls. CONCLUSIONS: Basophils of patients with CIU showed an activated profile, possibly due to an in vivo priming. Functionally, basophils have high responsiveness to IL-3 stimulation, thereby suggesting that defects in the signal transduction pathway after IgE cross-linking stimuli are recoverable in subjects with chronic urticaria.