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.

The present study compared the effects of follicle stimulating hormone (FSH) and luteinizing hormone (LH) on in vitro maturation (IVM), apoptosis, and secretion function in sheep oocytes, as well as gene expressions of the receptors (FSHR, LHR, and GnRHR) in cumulus-oocyte complexes (COCs). The COCs were recovered from sheep ovaries and pooled in groups. The COCs were cultured for 24 hours in IVM medium supplemented with various concentrations of LH (5-30 μg/mL) and FSH (5-30 IU/mL). They were allocated to LH-1 (5 µg/mL), LH-2 (10 µg/mL), LH-3 (20 µg/mL), and LH-4 (30 µg/mL) groups, and FSH-1 (5 IU/mL), FSH-2 (10 IU/mL), FSH-3 (20 IU/mL), and FSH-4 (30IU/mL) groups. The apoptosis of COCs was assessed by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). The maturation rates of oocytes improved gradually as LH and FSH concentration increased from 0 to 10 μg/mL(IU/mL), reaching a peak value of 44.1% of LH-2 and 48.5% of FSH-2 group. Oocyte apoptosis rates of LH-2 and FSH-2 groups were the lowest among LH- and FSH-treated groups, respectively. The germinal vesicle breakdown (GVBD) rate of the FSH-2 group was higher than that of the control group (CG) and FSH-4 groups. The GVBD rate of LH-2 group also increased in comparison with the CG group. FSH concentration of the FSH-2 group was greater than that of CG. Expression levels of FSHR, LHR, and GnRHR mRNAs of FSH-2, LH-3, and LH-3 group, respectively, were higher than CG. Levels of FSHR proteins in FSH-2 and FSH-3 groups were greater than CG. Levels of GnRHR proteins were increased with a maximum increment of FSH-4. The FSH and LH supplemented into the IVM medium could promote the maturation rate, reduce the apoptosis rate of sheep oocytes, and increase FSH concentrations in IVM medium fluid. Additionally, FSH and LH enhanced expression levels of FSHR, LHR, and GnRHR mRNAs of sheep COCs.

Oxidative stress plays an important role in the development of diabetic cardiomyopathy. In the present study, we determined whether the effect of astragalus polysaccharides (APS) on diabetic cardiomyopathy was associated with its impact on oxidative stress. Streptozotocin (STZ)-induced diabetic mice and heterozygous superoxide dismutase (SOD2+/-) knockout mice were administered APS. The hemodynamics, cardiac ultrastructure, and the apoptosis, necrosis and proliferation of cardiomyocytes were assessed to evaluate the effect of APS on diabetic and oxidative cardiomyopathy. Furthermore, H2O2 formation, oxidative stress/damage, and SOD activity in cardiomyocytes were evaluated to determine the effects of APS on cardiac oxidative stress. APS therapy improved hemodynamics and myocardial ultrastructure with reduced apoptosis/necrosis, and enhanced proliferation in cardiomyocytes from both STZ-induced diabetic mice and heterozygous SOD2+/- knockout mice. In addition, APS therapy reduced H2O2 formation and oxidative stress/damage, and enhanced SOD activity in both groups of mice. Our findings suggest that APS had benefits in diabetic cardiomyopathy, which may be partly associated with its impact on cardiac oxidative stress.

As atuais medições bidimensionais (2D) com marcadores ultrassônicos com inerentes intra- e interobservadores têm limitações de variabilidade. O objetivo deste trabalho é investigar o desempenho de medições convencionais 2D com marcador ultrassom e uma proposta interativa tridimensional programável de avaliação com marcador (3D). Isto é essencialmente importante para analisar que a medição em 3D volumétrica possui maior impacto e medição de reprodutibilidade vis-à-vis a 2D. Vinte e três casos de ultrassonografia pré-natal foram obtidos do hospital que colaborou com a pesquisa após a aprovação pela Comissão de Ética. A medida do marcador 2D ultrassônica é Translucência Nucal ou comumente abreviado como NT. A análise descritiva de medição de ultrassom 2D e 3D foram calculados. Três medidas foram obtidas em ensaios para cada método. Ambos os dados foram testados com uma amostra de teste de Kolmogorov-Smirnov e os resultados indicam que as medições de marcadores foram distribuídos normalmente com significativos valores paramétricos de 0, 621 e 0, 596, respectivamente. A média e o desvio padrão para ambos os métodos de medição são 1, 4495 ± 0, 46490 (2D) e 1, 3561 ± 0, 50994 (3D).O teste ANOVA mostrou que medições computadorizadas 3D foram insignificantemente diferentes da média 2D convencional com nível de significância de 0, 05. Com coeficiente de correlação de Pearson com valor R = 0, 861, o resultado comprova forte correlação linear positiva entre medidas ultrassônicas 2D e 3D. Reprodutibilidade e exatidão do ultrassom 3D na medida da TN foram significativamente aumentadas em comparação com a avaliação ultrassom 2D na modalidade B-pré-natal. Imagem reconstruída em 3D tem maiores valores clínicos em comparção com as imagens de ultrassom 2D que possuem menos informações para diagnóstico.

Current two-dimensional (2D) ultrasonic marker measurements are inherent with intra- and inter-observer variability limitations. The objective of this paper is to investigate the performance of conventional 2D ultrasonic marker measurements and proposed programmable interactive three-dimensional (3D) marker evaluation. This is essentially important to analyze that the measurement on 3D volumetric measurement possesses higher impact and reproducibility vis-à-vis 2D measurement. Twenty three cases of prenatal ultrasound examination were obtained from collaborating hospital after Ethical Committee's approval. The measured 2D ultrasonic marker is Nuchal Translucency or commonly abbreviated as NT. Descriptive analysis of both 2D and 3D ultrasound measurement were calculated. Three trial measurements were taken for each method. Both data were tested with One-Sample Kolmogorov-Smirnov Test and results indicate that markers measurements were distributed normally with significant parametric values at 0.621 and 0.596 respectively. Computed mean and standard deviation for both measurement methods are 1.4495 ± 0.46490 (2D) and 1.3561 ± 0.50994 (3D). ANOVA test shows that computerized 3D measurements were found to be insignificantly different from the mean of conventional 2D at the significance level of 0.05. With Pearson's correlation coefficient value or R = 0.861, the result proves strong positive linear correlation between 2D and 3D ultrasonic measurements. Reproducibility and accuracy of 3D ultrasound in NT measurement was significantly increased compared with 2D B-mode ultrasound prenatal assessment. 3D reconstructed imaging has higher clinical values compare to 2D ultrasound images with less diagnostics information.