Characterization of nontypable haemophilus influenzae isolates recovered from adult patients with underlying chronic lung disease reveals genotypic and phenotypic traits associated with persistent infection

Nontypable Haemophilus influenzae (NTHi) has emerged as an important opportunistic pathogen causing infection in adults suffering obstructive lung diseases. Existing evidence associates chronic infection by NTHi to the progression of the chronic respiratory disease, but specific features of NTHi associated with persistence have not been comprehensively addressed. To provide clues about adaptive strategies adopted by NTHi during persistent infection, we compared sequential persistent isolates with newly acquired isolates in sputa from six patients with chronic obstructive lung disease. Pulse field gel electrophoresis (PFGE) identified three patients with consecutive persistent strains and three with new strains. Phenotypic characterisation included infection of respiratory epithelial cells, bacterial self-aggregation, biofilm formation and resistance to antimicrobial peptides (AMP). Persistent isolates differed from new strains in showing low epithelial adhesion and inability to form biofilms when grown under continuous-flow culture conditions in microfermenters. Self-aggregation clustered the strains by patient, not by persistence. Increasing resistance to AMPs was observed for each series of persistent isolates; this was not associated with lipooligosaccharide decoration with phosphorylcholine or with lipid A acylation. Variation was further analyzed for the series of three persistent isolates recovered from patient 1. These isolates displayed comparable growth rate, natural transformation frequency and murine pulmonary infection. Genome sequencing of these three isolates revealed sequential acquisition of single-nucleotide variants in the AMP permease sapC, the heme acquisition systems hgpB, hgpC, hup and hxuC, the 3-deoxy-D-manno-octulosonic acid kinase kdkA, the long-chain fatty acid transporter ompP1, and the phosphoribosylamine glycine ligase purD. Collectively, we frame a range of pathogenic traits and a repertoire of genetic variants in the context of persistent infection by NTHi.

Pressão arterial e concentração plasmática do peptídeo atrial natriurético e do peptídeo natriurético tipo B, em gestações complicadas pela pré-eclâmpsia

OBJETIVO: o estudo busca determinar a existência de associação entre a elevação da pressão arterial e os níveis plasmáticos dos peptídeos natriuréticos ANP e BNP, na gestação complicada pela pré-eclâmpsia. MÉTODOS: em estudo transversal caso-controle, pareado por idade gestacional, 25 grávidas normotensas e 61 portadoras de pré-eclâmpsia foram avaliadas quanto ao nível da pressão arterial e às concentrações plasmáticas dos peptídeos natriuréticos ANP e BNP. Exames clínico e laboratoriais foram realizados para o diagnóstico de pré-eclâmpsia, sendo a pressão arterial medida de forma não invasiva. As dosagens hormonais foram obtidas por radioimunoensaio, após extração em colunas Sep-pak C18. Os valores médios das concentrações plasmáticas do ANP e BNP foram comparados entre grupos com pressão arterial progressivamente maiores. A correlação entre os valores da pressão arterial e os níveis plasmáticos do ANP e BNP no sangue materno foi também investigada pela de análise de regressão no grupo completo de gestantes e em grupos específicos excluindo-se a hipertensão anterior à gestação e, em seguida, excluindo-se aquelas sem hipertensão prévia. RESULTADOS: os valores plasmáticos de ANP foram 41.5±7.3, 78.4±13.1 e 89.2±13.4 pg/mL (p<0,00001) e os de BNP plasmático foram 79.5±15.8, 176.7±42.2 e 208.3±63.5 pg/mL (p=0,005), respectivamente, para os grupos de pressão arterial média =107 mmHg, 107-139 mmHg e =140 mmHg. Verificou-se correlação positiva entre as concentrações plasmáticas do ANP e os níveis pressóricos na pré-eclâmpsia, independente da existência de estado hipertensivo prévio à gestação (p<0,0001 para pré-eclâmpsia e p<0,01 para pré-eclampsia sobreposta à hipertensão arterial crônica), ao passo que as dosagens de BNP não se mostraram associadas à pressão arterial no grupo com hipertensão arterial prévia à gestação (p=0,004 para pré-eclâmpsia e p=0,18 para pré-eclampsia sobreposta à hipertensão arterial crônica). CONCLUSÃO: o agravamento da hipertensão na pré-eclâmpsia correlacionou-se com as concentrações séricas do ANP e BNP, embora os valores do BNP possam ser influenciados pela existência de estado hipertensivo prévio.

Antimicrobial resistance among invasive Haemophilus influenzae strains: results of a Brazilian study carried out from 1996 through 2000

A total of 1712 strains of Haemophilus influenzae isolated from patients with invasive diseases were obtained from ten Brazilian states from 1996 to 2000. ß-Lactamase production was assessed and the minimum inhibitory concentrations (MIC) of ampicillin, chloramphenicol, ceftriaxone and rifampin were determined using a method for broth microdilution of Haemophilus test medium. The prevalence of strains producing ß-lactamase ranged from 6.6 to 57.7%, with an overall prevalence of 18.4%. High frequency of ß-lactamase-mediated ampicillin resistance was observed in Distrito Federal (25%), São Paulo (21.7%) and Paraná (18.5%). Of the 1712 strains analyzed, none was ß-lactamase negative, ampicillin resistant. A total of 16.8% of the strains were resistant to chloramphenicol, and 13.8% of these also presented resistance to ampicillin, and only 3.0% were resistant to chloramphenicol alone. All strains were susceptible to ceftriaxone and rifampin and the MIC90 were 0.015 µg/ml and 0.25 µg/ml, respectively. Ceftriaxone is the drug of choice for empirical treatment of bacterial meningitis in pediatric patients who have not been screened for drug susceptibility. The emergence of drug resistance is a serious challenge for the management of invasive H. influenzae disease, which emphasizes the fundamental role of laboratory-based surveillance for antimicrobial resistance.

Aislamiento y caracterización de compuestos de plantas del noreste de México con actividad contra cepas de Streptococcus pneumoniae, Staphylococcus aureus y Haemophilus influenzae

Tesis (Doctorado en Ciencias con Especialidad en Química Biomédica) UANL

Determinación de genes regulados por microRNAs, involucrados en la leucemia linfocítica aguda tipo B (LLA-B).

Tesis (Doctorado en Ciencias con Orientación en Biología Molecular e Ingeniería Genética) UANL, 2013.

Household survey of hepatitis B vaccine coverage among Brazilian children

We conducted a multi-stage household cluster survey to calculate hepatitis B vaccine coverage among children 18-30 months of age in 27 Brazilian cities. Hepatitis B vaccine is administered at birth, 1 month and 6 months of age by Brazil`s national immunization program. Among 17,749 children surveyed, 40.2% received a birth dose within one day of birth, 94.8% received at least one dose of hepatitis B vaccine, and 86.7% completed the three-dose series by 12 months of age. Increased coverage with the birth dose and administration of hepatitis B in combination with diphtheria-tetanus-pertussis-Haemophilus influenzae type b antigens could improve protection against hepatitis B. (C) 2009 Elsevier Ltd. All rights reserved.

Estudo do índice crioscópico do leite tipo B "in natura" produzido na bacia leiteira do Vale do Taquari, RS.

O índice crioscópico é um dos parâmetros analíticos de precisão utilizados para determinar a qualidade físico-química do leite. É um valor diretamente ligado ao extrato seco do leite, mais especificamente em relação a presença, maior ou menor, de lactose e cloretos. A adição de água ao leite, como mecanismo de fraude, altera os valores do índice crioscópico. Em virtude disto, o mesmo é utilizado como um dos critérios para desclassificação de leite. O problema se estabelece quando os padrões determinados em legislação não condizem com as características reais do leite produzido. O presente trabalho objetivou comparar os resultados do índice crioscópico do leite tipo B “in natura” da bacia leiteira do Vale do Taquari com o valor estabelecido na legislação vigente, bem como observou as alterações deste parâmetro ao longo de um ano. O projeto foi desenvolvido no período de março de 2001 a fevereiro de 2002, com a participação de 10 propriedades produtoras de leite tipo B, onde as coletas foram realizadas mensalmente, perfazendo um total de 573 amostras no período. Os resultados analíticos foram obtidos mediante as análises de acidez, temperatura, densidade, gordura, extrato seco total (EST), extrato seco desengordurado (ESD) e índice crioscópico (IC). Quanto ao comportamento do índice crioscópico, o diagnóstico confirmou a necessidade imprescindível da implementação de parâmetros legais regionais, respeitando as características específicas de cada região. A pesquisa apresentou para o leite tipo B “in natura” do Vale do Taquari, um índice crioscópico médio de –0,537 °H, enquadrando-se no parâmetro estabelecido pela Instrução Normativa Nº 51 de 18/09/02, ou seja, máximo de –0,530 °H. Este índice também apresentou Valor médio mais baixo nos meses de junho e julho e as variações mensais individuais foram significativas. Estes comportamentos indicam que a alimentação ofertada ao rebanho influencia nestas alterações.

Peptídeo natriurético tipo-B como ferramento no manejo de insuficiência cardíaca e fibrilação atrial persistente

Resumo não disponível.

Comportamento dos níveis plasmáticos do Peptídeo Natriurético tipo-B em idosos com função sistólica do ventrículo esquerdo preservada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento, aperfeiçoamento e validação de método cromatográfico para previsão da qualidade e propriedades físico-químicas do óleo diesel tipo B

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Surveillance of antimicrobial resistance of Streptococcus pneumoniae and Haemophilus influenzae in children with pneumonia

In order to identify optimal therapy for children with bacterial pneumonia, Pakistan's ARI Program, in collaboration with the National Institute of Health (NIH), Islamabad, undertook a national surveillance of antimicrobial resistance in S. pneumoniae and H. influenzae. The project was carried out at selected urban and peripheral sites in 6 different regions of Pakistan, in 1991–92. Nasopharyngeal (NP) specimens and blood cultures were obtained from children with pneumonia diagnosed in the outpatient clinic of participating facilities. Organisms were isolated by local hospital laboratories and sent to NIH for confirmation, serotyping and antimicrobial susceptibility testing. Following were the aims of the study (i) to determine the antimicrobial resistance patterns of S. pneumoniae and H. influenzae in children aged 2–59 months; (ii) to determine the ability of selected laboratories to identify and effectively transport isolates of S. pneumoniae and H. influenzae cultured from nasopharyngeal and blood specimens; (iii) to validate the comparability of resistance patterns for nasopharyngeal and blood isolates of S. pneumoniae and H. influenzae from children with pneumonia; and (iv) to examine the effect of drug resistance and laboratory error on the cost of effectively treating children with ARI. ^ A total of 1293 children with ARI were included in the study: 969 (75%) from urban areas and 324 (25%) from rural parts of the country. Of 1293, there were 786 (61%) male and 507 (39%) female children. The resistance rate of S. pneumoniae to various antibiotics among the urban children with ARI was: TMP/SMX (62%); chloramphenicol (23%); penicillin (5%); tetracycline (16%); and ampicillin/amoxicillin (0%). The rates of resistance of H. influenzae were higher than S. pneumoniae: TMP/SMX (85%); chloramphenicol (62%); penicillin (59%); ampicillin/amoxicillin (46%); and tetracycline (100%). There were similar rates of resistance to each antimicrobial agent among isolates from the rural children. ^ Of a total 614 specimens that were tested for antimicrobial susceptibility, 432 (70.4%) were resistant to TMP/SMX and 93 (15.2%) were resistant to antimicrobial agents other than TMP/SMX viz. ampicillin/amoxicillin, chloramphenicol, penicillin, and tetracycline. ^ The sensitivity and positive predictive value of peripheral laboratories for H. influenzae were 99% and 65%, respectively. Similarly, the sensitivity and positive predictive value of peripheral laboratory tests compared to gold standard i.e. NIH laboratory, for S. pneumoniae were 99% and 54%, respectively. ^ The sensitivity and positive predictive value of nasopharyngeal specimens compared to blood cultures (gold standard), isolated by the peripheral laboratories, for H. influenzae were 88% and 11%, and for S. pneumoniae 92% and 39%, respectively. (Abstract shortened by UMI.)^

Desarrollo de software de control y medida para goniofotómetro de tipo B

El proyecto trata del desarrollo de un software para realizar el control de la medida de la distribución de intensidad luminosa en luminarias LED. En el trascurso del proyecto se expondrán fundamentos teóricos sobre fotometría básica, de los cuales se extraen las condiciones básicas para realizar dicha medida. Además se realiza una breve descripción del hardware utilizado en el desarrollo de la máquina, el cual se basa en una placa de desarrollo Arduino Mega 2560, que, gracias al paquete de Labview “LIFA” (Labview Interface For Arduino”), será posible utilizarla como tarjeta de adquisición de datos mediante la cual poder manejar tanto sensores como actuadores, para las tareas de control. El instrumento de medida utilizado en este proyecto es el BTS256 de la casa GigaHerzt-Optik, del cual se dispone de un kit de desarrollo tanto en lenguaje C++ como en Labview, haciendo posible programar aplicaciones basadas en este software para realizar cualquier tipo de adaptación a las necesidades del proyecto. El software está desarrollado en la plataforma Labview 2013, esto es gracias a que se dispone del kit de desarrollo del instrumento de medida, y del paquete LIFA. El objetivo global del proyecto es realizar la caracterización de luminarias LED, de forma que se obtengan medidas suficientes de la distribución de intensidad luminosa. Los datos se recogerán en un archivo fotométrico específico, siguiendo la normativa IESNA 2002 sobre formato de archivos fotométricos, que posteriormente será utilizado en la simulación y estudio de instalaciones reales de la luminaria. El sistema propuesto en este proyecto, es un sistema basado en fotometría tipo B, utilizando coordenadas VH, desarrollando un algoritmo de medida que la luminaria describa un ángulo de 180º en ambos ejes, con una resolución de 5º para el eje Vertical y 22.5º para el eje Horizontal, almacenando los datos en un array que será escrito en el formato exigido por la normativa. Una vez obtenidos los datos con el instrumento desarrollado, el fichero generado por la medida, es simulado con el software DIALux, obteniendo unas medidas de iluminación en la simulación que serán comparadas con las medidas reales, intentando reproducir en la simulación las condiciones reales de medida. ABSTRACT. The project involves the development of software for controlling the measurement of light intensity distribution in LEDs. In the course of the project theoretical foundations on basic photometry, of which the basic conditions for such action are extracted will be presented. Besides a brief description of the hardware used in the development of the machine, which is based on a Mega Arduino plate 2560 is made, that through the package Labview "LIFA" (Interface For Arduino Labview "), it is possible to use as data acquisition card by which to handle both sensors and actuators for control tasks. The instrument used in this project is the BTS256 of GigaHerzt-Optik house, which is available a development kit in both C ++ language as LabView, making it possible to program based on this software applications for any kind of adaptation to project needs. The software is developed in Labview 2013 platform, this is thanks to the availability of the SDK of the measuring instrument and the LIFA package. The overall objective of the project is the characterization of LED lights, so that sufficient measures the light intensity distribution are obtained. Data will be collected on a specific photometric file, following the rules IESNA 2002 on photometric format files, which will then be used in the simulation and study of actual installations of the luminaire. The proposed in this project is a system based on photometry type B system using VH coordinates, developing an algorithm as the fixture describe an angle of 180 ° in both axes, with a resolution of 5 ° to the vertical axis and 22.5º for the Horizontal axis, storing data in an array to be written in the format required by the regulations. After obtaining the data with the instrument developed, the file generated by the measure, is simulated with DIALux software, obtaining measures of lighting in the simulation will be compared with the actual measurements, trying to play in the simulation the actual measurement conditions .

Human milk lactoferrin inactivates two putative colonization factors expressed by Haemophilus influenzae

Haemophilus influenzae is a major cause of otitis media and other respiratory tract disease in children. The pathogenesis of disease begins with colonization of the upper respiratory mucosa, a process that involves evasion of local immune mechanisms and adherence to epithelial cells. Several studies have demonstrated that human milk is protective against H. influenzae colonization and disease. In the present study, we examined the effect of human milk on the H. influenzae IgA1 protease and Hap adhesin, two autotransported proteins that are presumed to facilitate colonization. Our results demonstrated that human milk lactoferrin efficiently extracted the IgA1 protease preprotein from the bacterial outer membrane. In addition, lactoferrin specifically degraded the Hap adhesin and abolished Hap-mediated adherence. Extraction of IgA1 protease and degradation of Hap were localized to the N-lobe of the bilobed lactoferrin molecule and were inhibited by serine protease inhibitors, suggesting that the lactoferrin N-lobe may contain serine protease activity. Additional experiments revealed no effect of lactoferrin on the H. influenzae P2, P5, and P6 outer-membrane proteins, which are distinguished from IgA1 protease and Hap by the lack of an N-terminal passenger domain or an extracellular linker region. These results suggest that human milk lactoferrin may attenuate the pathogenic potential of H. influenzae by selectively inactivating IgA1 protease and Hap, thereby interfering with colonization. Future studies should examine the therapeutic potential of lactoferrin, perhaps as a supplement in infant formulas.

Haemophilus influenzae pili are composite structures assembled via the HifB chaperone.

Haemophilus influenzae is a Gram-negative bacterium that represents a common cause of human disease. Disease due to this organism begins with colonization of the upper respiratory mucosa, a process facilitated by adhesive fibers called pili. In the present study, we investigated the structure and assembly of H. influenzae pili. Examination of pili by electron microscopy using quick-freeze, deep-etch and immunogold techniques revealed the presence of two distinct subassemblies, including a flexible two-stranded helical rod comprised of HifA and a short, thin, distal tip structure containing HifD. Genetic and biochemical studies demonstrated that the biogenesis of H. influenzae pili is dependent on a periplasmic chaperone called HifB, which belongs to the PapD family of immunoglobulin-like chaperones. HifB bound directly to HifA and HifD, forming HifB-HifA and HifB-HifD complexes, which were purified from periplasmic extracts by ion-exchange chromatography. Continued investigation of the biogenesis of H. influenzae pili should provide general insights into organelle development and may suggest novel strategies for disease prevention.