Resumo Infecções do trato respiratório inferior (ITRIs) são as infecções mais comuns entre pacientes internados em unidade de terapia intensiva (UTI). Pseudomonas aeruginosa é a causa mais comum de ITRIs e está associada ao aumento da mortalidade. Diferentes espécies de Pseudomonas exibem diferentes perfis genéticos e resistência variada as drogas. O presente estudo determina a epidemiologia molecular através do método de fingerprinting de DNA e resistência as drogas de P. aeruginosa isoladas de pacientes com LTRIs internados em UTI. Um total de 79 P. aeruginosa isoladas de pacientes com ITRIs internados em UTI foram caracterizados por Polimorfismo de Comprimento de Fragmentos de Restrição (RFLP), DNA Polimórfico Amplificado ao Acaso (RAPD) e PCR Extrapalindrômico Repetitivo (REP-PCR). A resistência aos antibióticos foram determinadas pelos ensaios de concentrações inibitória mínima (MIC), enquanto os genes MDR, blaTEM, blaOXA, blaVIM, blaCTX-M-15 foram detectados pela reação em cadeia da polimerase (PCR). Das 137 Pseudomonas sp isoladas de pacientes de UTI, 57,7% dos isolados foram relatados como P. aeruginosa. A prevalência geral de P. aeruginosa entre os pacientes incluídos foram de 34,5%. A análise RAPD renderam 45 padrões diferentes com 72 clusters com nível de similaridade de 57% a 100%. A análise RFLP renderam 8 padrões diferentes com 14 clusters com 76% a 100% de similaridade. A análise de PCR do REP produziram 37 padrões diferentes com 65 clusters com nível de similaridade de 56% a 100%. Não houveram correlações entre os diferentes padrões de DNA observados entre os três diferentes métodos. Predominantes dos isolados (46,8%) eram resistentes à amicacina. Dos 79 isolados, 60,8% foram positivos para o gene blaTEM e 39,2% foram positivos para o gene blaOXA. P. aeruginosa foi predominantemente isolado de pacientes com ITRIs internados em UTI. A diferença no nível de similaridade observado entre os três métodos de fingerprinting do DNA indica que há alta variabilidade inter-strain. A alta variabilidade genética e os padrões de resistência indicam que devemos monitorar continuamente a tendência na prevalência e resistência a antibióticos de P. aeruginosa, especialmente em pacientes com ITRIs internados em UTI.

Abstract Lower respiratory tract infections (LRTIs) caused by Pseudomonas aeruginosa are the most common infection among hospitalized patients, associated with increased levels of morbidity, mortality and attributable health care costs. Increased resistant Pseudomonas worldwide has been quite meaningful to patients, especially in intensive care unit (ICUs). Different species of Pseudomonas exhibit different genetic profile and varied drug resistance. The present study determines the molecular epidemiology through DNA fingerprinting method and drug resistance of P. aeruginosa isolated from patients with LTRIs admitted in ICU. A total of 79 P. aeruginosa isolated from patients with LRTIs admitted in ICU were characterized by Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD) and Repetitive Extrapalindromic PCR (REP-PCR). Antibiotic resistance was determined by minimum inhibitory concentration (MIC) assay while MDR genes, viz, blaTEM, blaOXA, blaVIM, blaCTX-M-15 were detected by polymerase chain reaction (PCR). Of the 137 Pseudomonas sp isolated from ICU patients, 57.7% of the isolates were reported to be P. aeruginosa. The overall prevalence of P. aeruginosa among the all included patients was 34.5%. The RAPD analysis yielded 45 different patterns with 72 clusters with 57% to 100% similarity level. The RFLP analysis yielded 8 different patterns with 14 clusters with 76% to 100% similarity level. The REP PCR analysis yielded 37 different patterns with 65 clusters with 56% to 100% similarity level. There was no correlation among the different DNA patterns observed between the three different methods. Predominant of the isolates (46.8%) were resistant to amikacin. Of the 79 isolates, 60.8% were positive for blaTEM gene and 39.2% were positive for blaOXA gene. P. aeruginosa was predominantly isolated from patients with LRTIs admitted in ICU. The difference in the similarity level observed between the three DNA fingerprinting methods indicates that there is high inter-strain variability. The high genetic variability and resistance patterns indicates that we should continuously monitor the trend in the prevalence and antibiotic resistance of P. aeruginosa especially in patients with LRTIs admitted in ICU.

RESUMO Infecção com o vírus Hepatite E (HEV) é uma importante questão de saúde pública global. Infecções HEV são reconhecidas como doença zoológica. Acredita-se que suínos são o principal reservatório de HEV. Recentemente iaques, vacas, e gado amarelo foram reportados como novos reservatórios do HEV. Porém, não se sabe se outras espécies de gado e búfalo são sensíveis a infecção HEV. Para investigar a prevalência de infecção HEV em búfalos, foram realizados prova de imunoabsorção enzimática e polimerização em cadeia inversa ancorada em transcrição. Apenas um búfalo foi positivo para o anticorpo anti-HEV IgM (1/106, 0,94%), e nenhum foi positivo para o anticorpo anti-HEV IgG. Para nossa surpresa cinco (5/106, 4,72%) e três amostras de leite (3/40, 7,50%) de búfalos foram positivos para HEV RNA. Todas as estirpes de HEV isoladas de búfalos pertencem ao genótipo 4. Resultados indicam que búfalos podem ser um reservatório de HEV.

ABSTRACT Hepatitis E virus (HEV) infection is an important global public health issue. HEV infections are recognized as a zoonotic disease. Swine are believed to be the main reservoir of HEV. Recently, yaks, cows, and yellow cattle have been reported as new reservoirs of HEV. However, whether other species of cattle and buffaloes are sensitive to HEV infection is unknown. To investigate the prevalence of HEV infection in buffaloes, enzyme-linked immunosorbent assay (ELISA) and reverse transcription-nested polymerase chain reaction (RT-nPCR) were performed. Only one buffalo was positive to anti-HEV IgM antibody (1/106, 0.94%), and none were positive for anti-HEV IgG antibody. To our surprise, five serum (5/106, 4.72%) and three milk samples (3/40, 7.50%) from buffaloes were positive to HEV RNA. All strains of HEV isolated from buffaloes belong to genotype 4. Results indicate that buffaloes may be a new reservoir of HEV.

RESUMO Nesse estudo, folhas "neem" foram extraídas com sucesso com éter de petróleo, 95% de etanol e água, e as atividades inseticidas desses extratos foram medidas contra larvas de Oxya chinesis. Os resultados mostram que extrato com 95% de etanol deram o maior resultado de extração e atividade inseticida e foi então extraído utilizando mais cinco diferentes solventes. O éter de petróleo do extrato de 95% etanol apresentou maior atividade inseticida com concentração letal média variando de 14.93 a 55.66mg/mL. A análise por cromatografia de massa mostrou que o extrato de éter de petróleo está composto principalmente de alcanos, alcenos, ésteres e amidas. A avaliação patológica revelou que as lesões proeminentes, inclusive células regenerativas reduzidas no intestino e células cilíndricas edemaciadas e degeneradas foram observadas no quinto estágio de desenvolvimento da Oxya chinesis após tratamento. As características ultraestruturais mostraram que as células cilíndricas, microvilos e mitocôndrias apresentavam lesões graves. Esses resultados sugerem que o extrato de éter de petróleo de folhas de "neem" tem atividade inseticida potente e pode ser um candidato a inseticida.

ABSTRACT In this study, neem leaves were successively extracted with petroleum ether, 95% ethanol and water and the insecticidal activities of these extracts against Oxya chinensis larvae were measured. The results showed that 95% ethanol extract gave the highest extraction yield and insecticidal activity, and it was further extracted with five different solvents. The petroleum ether extract from the 95% ethanol extract possessed the highest insecticidal activity with median lethal concentration values ranging from 14.93 to 55.66mg/mL. The gas chromatography-mass spectrometer analysis showed that the petroleum ether extract mainly composed of alkanes, olefin, esters and amide. The pathological examination revealed that the prominent lesions, including reduced regenerative cells in midgut and swelled and degenerated cylindrical cells, were observed in the 5th instar Oxya chinensis after treatment. The ultrastructural features showed that the cylindrical cells, microvilli and mitochondria were seriously damaged. These results suggested that the petroleum ether extract from neem leaves had potent insecticidal activity and could be a candidate insecticide.

To reduce production costs and increase efficiency in ageing coal mines, a system that utilizes two shearers in a fully mechanized longwall working face is proposed. Using theoretical and engineering experience, three issues pertaining to the system were determined: (a) the two-shearer mining technique; (b) matching and modifying the equipment; (c) and coal-cutting task allocation for each shearer. Two mining processes are proposed, with the two shearers travelling in either the same or opposing directions. To avoid breakage of chains and pan extrusion of the armoured face conveyor (AFC), the length of the AFC 'snake' was controlled. To ensure continuous cutting and transporting of coal, the cross-sectional dimensions of the AFC were checked and the shearer closer to the head drive was modified. Based on the assumption of equal cutting times for each shearer, the meeting position of the two shearers was obtained by using a theoretical model of mining task allocation. Application of this technique at No. 2 Jining Mine, China, has shown remarkable benefits: daily production capacity was increased by 54% and personnel efficiency was improved by 33 t/d per person.

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.

Epidemiological studies of short and long sleepers have not been conducted previously. We collected socioeconomic, psychological, and polysomnographic characteristics of 6501 parents (3252 men and 3249 women) of 4036 primary school children in Guangzhou city. The study data were collected in three phases. The overall prevalence of short (5 h or less) and long (10 h or more) sleep duration was 0.52 and 0.64%, respectively. Long sleepers had higher Eysenck Personality Questionnaire neuroticism scores [odds ratio (OR)=1.224, 95% confidence interval (CI)=1.047-1.409] and lower education levels (OR=0.740, 95%CI=0.631-0.849) than short sleepers. In the polysomnographic assessment, short, long, and normal sleepers (7-8 h) shared similar durations of Stage 3 sleep (short=25.7±10.7, long=20.3±7.9, and normal=28.0±12.8 min, F=1.402, P=0.181). In daytime multiple sleep latency tests, short sleepers (10/19, 52.6%) were more prone to have a short sleep latency (≤8 min) than long sleepers (2/23, 8.7%). In addition to different sleep durations, neuroticism might also contribute to differences between short and long sleepers in social achievements. Stage 3 sleep might be essential for humans. The short sleep latency (≤8 min) of short sleepers in multiple sleep latency tests should be interpreted cautiously, since it was of the same severity as required for a diagnosis of narcolepsy or idiopathic hypersomnia.

Porous silicon (PSi) has attracted much attention for biosensing due to its large surface area and easy preparation. In this paper, an optical immunoassay method based on a macro-PSi silicon resonant waveguide, with pore size much larger than that of large molecules such as bovine serum albumin, has been developed for the detection of the antigen-antibody reaction between hydroxysafflor yellow A (HSYA, i.e., the component of Carthamus tinctorius L.) and the specificity of the polyclonal anti-HSYA antibodies. HSYA antibodies were immobilized into the macro-PSi silicon waveguide using standard amino-silane and glutaraldehyde chemistry. The waveguide resonance angle was increased by binding HSYA onto the immobilized antibodies. The label-free immunosensor is simple and exhibits high sensitivity to HSYA. Therefore, this research is expected to have applications for quick and accurate determination of HSYAand can also be used for various immunoassays with other antigens..