Viability of enteric bacterial pathogens in transport media

Bacterial pathogens such as enterotoxigenic Escherichia coli, Salmonella, and Campylobacter spp. are associated with up to 80% of diarrheal illness to travelers from developed countries to developing countries. In order to study acute gastrointestinal diseases, researchers from developed countries such as the United States rely on transporting clinical specimens from the developing countries to laboratories in the U.S. in transport media systems. There are few commercially available transport media systems cited in the literature or designated by transport system manufacturers for the transport of enteric bacteria. Therefore a laboratory-based study was conducted to assess three commercial available transport media systems, two gel swabs and one liquid vial, to determine the most appropriate for the maintenance and recovery of common enteric bacterial pathogens. A total of 13 bacterial enteropathogens were recovered from 25°C and 4°C storage temperatures at time points up to 21 days. The results demonstrated that the gel swab and liquid vial transport systems performed similarly for all isolates at both temperatures. All three transport media systems struggled to maintain the isolates at recoverable concentrations when stored at 4°C and it is recommended that isolates be stored at 25°C in transport media systems. Lastly, swab transport systems are recommend for transport since they are small and easy to pack, resist leakage, and are less expensive than similarly performing liquid vial transport media systems.^

Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) nasal colonization among dental professionals

Background. MRSA (methicillin-resistant Staphylococcus aureus) is a multi-drug resistant bacterium that is quite prevalent in social environments where close person-to-person contact and crowding are an issue. In dental settings, the likelihood of transmission of MRSA may be higher than among other healthcare practitioners because of the close proximity between a patient's nose (where MRSA colonizes) and the field of procedure (the mouth) to the dental professional. Objective. To estimate the prevalence of MRSA nasal colonization among dental professionals (dentists and dental hygienists) in the Greater Houston Metropolitan Area, Texas, and analyze its associations with demographic, professional and personal protective equipment-related variables. Methods. 800 dental professionals (400 dentists and 400 dental hygienists) were randomly selected in the Greater Houston Metropolitan Area. Multiple waves of nasal swab kits and a self-administered questionnaire were mailed to increase the response rate of the study population. The swabs were cultured on chromagenic agar growth medium and bacterial growth results were evaluated after 18 hours. Positively selected bacterial colonies were confirmed as MRSA by further culturing these isolated bacteria on blood agar plates. Associations between positive nasal swabs and self-reported professional practice patterns, personal protective equipment use and demographics were analyzed using multiple logistic regression. Main Results. Completed questionnaires and nasal swabs were received from 496 study participants (68%). Fourteen cultures were positive for MRSA (4.2% among dentists and 1.6% among dental hygienists, p=0.07). After adjusting for gender, dental hygienists had a significantly lower prevalence of nasal colonization of MRSA as compared to dentists (OR: 0.20, 95% CI: 0.05–0.75). No other significant associations or interactions were found. Conclusion. The prevalence of nasal colonization with MRSA among dentists is similar to that reported for health care workers in general, whereas prevalence among dental hygienists is only slightly above that of the general population (1%). Differences in practice patterns and use of personal protective equipment did not explain this difference in this study, and was possibly due either to residual confounding or unexplored risk factors. Increased prevalence of MRSA among dentists warrants further investigation as to the reason for the increased rate and to allow implementation of measures to avoid transmission and progression to disease. ^

Influenza A (H1N1) con alteración de enzimas hepáticas en paciente vacunado

El compromiso hepático en la infección por virus Influenza A (H1N1) es muy infrecuente. Presentamos un caso de un paciente varón de 21 años, sano, vacunado, que consulta con un cuadro clásico de gripe. En el examen físico no se palpa hepatoesplenomegalia ni se observa ictericia. Se diagnostica gripe por virus de la influenza A (H1N1) y se inicia tratamiento con oseltamivir oral. Los exámenes de laboratorio revelaron elevación moderada de enzimas hepáticas. Los anticuerpos para virus de la hepatitis A, B y C, virus Epstein-Barr y citomegalovirus (CMV) fueron negativos. El hisopado nasofaríngeo fue positivo para influenza A (H1N1) con prueba de reacción en cadena de polimerasa en tiempo real (PCRRT). Se detectó hepatomegalia homogénea por ecografía abdominal. El cuadro clínico se resolvió en una semana, permaneciendo elevadas las enzimas hepáticas por 21 días. Discutimos los probables mecanismos de la injuria hepática en este caso.

A recombinant Chlamydia trachomatis major outer membrane protein binds to heparan sulfate receptors on epithelial cells.

Chlamydial attachment to columnar conjunctival or urogenital epithelial cells is an initial and critical step in the pathogenesis of chlamydial mucosal infections. The chlamydial major outer membrane protein (MOMP) has been implicated as a putative chlamydial cytoadhesin; however, direct evidence supporting this hypothesis has not been reported. The function of MOMP as a cytoadhesin was directly investigated by expressing the protein as a fusion with the Escherichia coli maltose binding protein (MBP-MOMP) and studying its interaction with human epithelial cells. The recombinant MBP-MOMP bound specifically to HeLa cells at 4 degrees C but was not internalized after shifting the temperature to 37 degrees C. The MBP-MOMP competitively inhibited the infectivity of viable chlamydiae for epithelial cells, indicating that the MOMP and intact chlamydiae bind the same host receptor. Heparan sulfate markedly reduced binding of the MBP-MOMP to cells, whereas chondroitin sulfate had no effect on binding. Enzymatic treatment of cells with heparitinase but not chondroitinase inhibited the binding of MBP-MOMP. These same treatments were also shown to reduce the infectivity of chlamydiae for epithelial cells. Mutant cell lines defective in heparan sulfate synthesis but not chondroitin sulfate synthesis showed a marked reduction in the binding of MBP-MOMP and were also less susceptible to infection by chlamydiae. Collectively, these findings provide strong evidence that the MOMP functions as a chlamydial cytoadhesin and that heparan sulfate proteoglycans are the host-cell receptors to which the MOMP binds.

Immunolocalization of the mercurial-insensitive water channel and glycerol intrinsic protein in epithelial cell plasma membranes.

Two water channel homologs were cloned recently from rat kidney, mercurial-insensitive water channel (MIWC) and glycerol intrinsic protein (GLIP). Polyclonal antibodies were raised against synthetic C-terminal peptides and purified by affinity chromatography. MIWC and GLIP antibodies recognized proteins in rat kidney with an apparent molecular mass of 30 and 27 kDa, respectively, and did not cross-react. By immunofluorescence, MIWC and GLIP were expressed together on the basolateral plasma membrane of collecting duct principal cells in kidney. By immunohistochemistry, MIWC and GLIP were expressed on tracheal epithelial cells with greater expression of GLIP on the basal plasma membrane and MIWC on the lateral membrane; only MIWC was expressed in bronchial epithelia. In eye, GLIP was expressed in conjunctival epithelium, whereas MIWC was found in iris, ciliary body, and neural cell layers in retina. MIWC and GLIP colocalized on the basolateral membrane of villus epithelial cells in colon and brain ependymal cells. Expression of MIWC and GLIP was not detected in small intestine, liver, spleen, endothelia, and cells that express water channels CHIP28 or WCH-CD. These studies suggest water/solute transporting roles for MIWC and GLIP in the urinary concentrating mechanism, cerebrospinal fluid absorption, ocular fluid balance, fecal dehydration, and airway humidification. The unexpected membrane colocalization of MIWC and GLIP in several tissues suggests an interaction at the molecular and/or functional levels.

Células-tronco de membrana amniótica de cão como terapia alternativa para o tratamento de ceratoconjuntivite seca em cães

A Ceratoconjuntivite Seca (KCS Keratoconjunctivitis Sicca) é uma desordem imunomediada e resulta de alterações do componente aquoso do filme lacrimal e da deficiência dos componentes lipídicos e mucoso.Seu diagnóstico é baseado no Teste Lacrimal de Schirmer (TLS) e no Teste de Ruptura do Filme Lacrimal (TRFL) e tem como sinais clínicos: secreção mucopurulenta, hiperemia conjuntival, blefaroespasmos, fotofobia, incômodo, dor, vascularização, opacidade corneana e pigmentação, além de cegueira em casos avançados. O tratamento convencional consiste em aplicações diárias de Ciclosporina 0,2% ou Tacrolimus 0,03% (pomada ou colírio oftálmicos), que apesar de controlar a doença, são custosos, não curativos e exigem alto comprometimento da interação paciente-proprietário. A terapia celular usando células-tronco (CT) traz uma nova esperança para doenças sem tratamento efetivo. Neste trabalho utilizamos CT mesenquimais (CTM) obtidas a partir de membrana amniótica (CTMA) de cães obtidas a partir do descarte destes tecidos em campanhas de castrações em diferentes tempos gestacionais, sem formação tumoral quando submetidas ao teste tumorigênico durante 60 dias. Dois animais com KCS crônica foram tratados com duas injeções de CTMA com intervalo de 30 dias, sendo a primeira de 0,5x106 células e a segunda de 1x106 células em cada glândula. Na segunda semana após a terapia foi observado aumento da TLS sugerindo um benéficio da terapia que foi diminuindo com o passar das semanas. O TRFL oscilou durante os testes e não apresentou diferenças significativas. A terapia celular utilizando CTMA de cães melhorou a condição ocular nos dois casos em momentos e parâmetros variados, com repercussão na melhoria da superfície, mas não houve regressão do quadro clínico. Investigações futuras em estágios menos avançados da doença podem ajudar a elucidar os mecanismos pelos quais esse efeito foi obtido

Estudo da viabilidade para introduzir na rotina testes de diagnóstico para infecção respiratória aguda

Para avaliar os benefícios da comunicação rápida ao clínico do diagnóstico de vírus respiratórios, foi analisado a viabilidade econômica de 2 testes, com o tempo de entrega de resultado em 2 horas para teste rápido e 48 horas para Biologia Molecular. As amostras coletadas foram processadas utilizando técnicas convencionais e os testes disponíveis no mercado local. Foram escolhidos dois testes rápidos pelo método de imunocromatografia para quatro parâmetros analíticos: Influenza A, Influenza H1N1, Influenza B e Vírus Sincicial Respiratório (RSV) e em Biologia Molecular um teste de RT-PCR multiplex com 25 patógenos entre vírus e bactérias. O tipo de amostra utilizada foi swab e lavado de nasofaringe. A população escolhida para o estudo foi paciente adulto, em tratamento de câncer, que necessita de uma resposta rápida já que a maioria se encontra com comprometimento do sistema imune por doença ou por tratamento. O estudo foi transversal, realizado entre os anos de 2012 e 2013, para avaliar a viabilidade econômica da introdução de testes de diagnóstico da infecção respiratória aguda de etiologia viral a partir de amostras de nasofaringe em pacientes com câncer atendidos no Centro de Atendimento de Oncologia Intercorrência (CAIO ), do Instituto do Câncer do Estado de São Paulo (ICESP), hospital público que atende exclusivamente Sistema Único de Saúde (SUS) e Hospital A.C. Camargo, que atende tanto a pacientes do SUS como da rede privada. O estudo incluiu 152 pacientes em tratamento para qualquer tipo de câncer, predominantemente do sexo feminino (81 mulheres e 70 homens) com idades entre 18-86 anos. Para participar do estudo o paciente era consultado e o critério para escolha do paciente foi ser portador de câncer, com história de febre (ainda que referida) acompanhada de tosse ou dor de garganta, tosse e sintomas respiratórios agudos, atendidos por protocolo padronizado que inclui avaliação na admissão, seguimento e manejo antimicrobiano. Para a avaliação econômica os pacientes foram classificados de acordo com o estado geral de saúde, se apresentavam bom estado de estado de saúde poderiam receber alta e faziam uso da medicação em casa evitando 5 dias de internação se recebessem algum resultado para Influenza ou RSV, no entanto os pacientes que apresentavam outro vírus, resultado negativo ou o estado geral era ruim permaneciam internados por 7 dias em observação e cuidados com medicação adequada. Foram realizadas análises econômicas em dois âmbitos: o sistema de saúde publico e o privado considerando o fator diminuição de dias de internação. A analise de Custo-benefício foi eficiente no Sistema privado mas inadequada para o SUS assim como, qualquer outra medida monetária já que os valores de reembolso do SUS estão defasados do custo de qualquer internação. A análise de Custo-efetividade que olha para outros fatores além do monetário foi efetiva nos dois sistemas que enfrentam falta de leitos além da condição de saúde do paciente de evitar a ingestão desnecessária de antibióticos, evitar os gastos do acompanhante, perda de dias de trabalho e estudo. Não houve correspondência de resultados dos testes rápidos com o multiplex de Biologia Molecular

Clinical Characterization of Asymptomatic or Minimally Symptomatic Young Patients Showing Signs Compatible With Dry Eye: A Pilot Study

Purpose: To evaluate and characterize the clinical profile of young asymptomatic or minimally symptomatic patients without diagnosis of dry eye but showing signs compatible with dry eye syndrome (DES). Methods: Prospective study including a total of 50 consecutive subjects with ages ranging from 18 to 40 years that were identified as asymptomatic or minimally symptomatic by means of the Ocular Surface Disease Index (OSDI) (score of <22). In all patients, a complete battery of tests for the diagnosis of DES was performed including the evaluation of the tear film break-up time (TFBUT), the level of corneal and conjunctival staining, and the eyelid and Meibomian morphology. Results: The OSDI score was significantly higher in women than in men (median: 12.5 vs. 5.3, P=0.01). Low grades of ocular surface staining, dysfunction of Meibomian gland expression, and alteration of quality of Meibomian secretions were observed in 56%, 58%, and 84% of eyes, respectively. More eyes with some dysfunction of Meibomian gland expressibility had a TFBUT less than 5 sec (P=0.033). A statistically significant difference in the OSDI score was found between patients with and without systemic allergies (P=0.036) and between male and female (P=0.01). Likewise, the OSDI score was significantly higher in those women wearing contact lenses compared with those not wearing them (P=0.012). Conclusions: Asymptomatic or minimally symptomatic young subjects may present low grades of clinical signs compatible with DES, with a trend to more symptomatology in women and allergic patients. These outcomes should be confirmed in future studies with larger samples.

Quando a hiperémia conjuntival constitui um desafio : a propósito de um caso clínico de tumor epidermóide da conjuntiva

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Monitoring influenza vaccine effectiveness using the national influenza surveillance system

Background: Flu vaccine composition is reformulated on a yearly basis. As such, the vaccine effectiveness (VE) from previous seasons cannot be considered for subsequent years, and it is necessary to monitor the VE for each season. This study (MonitorEVA- monitoring vaccine effectiveness) intends to evaluate the feasibility of using the national influenza surveillance system (NISS) for monitoring the influenza VE. Material and methods: Data was collected within NISS during 2004 to 2014 seasons. We used a case-control design where laboratory confirmed incident influenza like illness (ILI) patients (cases) were compared to controls (ILI influenza negative). Eligible individuals consisted on all aged individuals that consult a general practitioner or emergency room with ILI symptoms with a swab collected within seven days of symptoms onset. VE was estimated as 1- odds ratio of being vaccinated in cases versus controls adjusted for age and month of onset by logistic regression. Sensitivity analyses were conducted to test possible effect of assumptions on vaccination status, ILI definition and timing of swabs (<3 days after onset). Results: During the 2004-2014 period, a total of 5302 ILI patients were collected but 798 ILI were excluded for not complying with inclusion criteria. After data restriction the sample size in both groups was higher than 148 individuals/ season; minimum sample size needed to detect a VE of at least 50% considering a level of significance of 5% and 80% power. Crude VE point estimates were under 45% in 2004/05, 2005/06, 2011/12 and 2013/14 season; between 50%-70% in 2006/07, 2008/09 and 2010/11 seasons, and above 70% in 2007/08 and 2012/13 season. From season 2006/07 to 2013/14, all crude VE estimates were statistically significant. After adjustment for age group and month of onset, the VE point estimates decreased and only 2008/09, 2012/13 and 2013/14 seasons were significant. Discussion and Conclusions: MonitorEVA was able to provide VE estimates for all seasons, including the pandemic, indicating if the VE was higher than 70% and less than 50%. When comparing with other observational studies, MonitorEVA estimates were comparable but less precise and VE estimates were in accordance with the antigenic match of the circulating virus/ vaccine strains. Given the sensitivity results, we propose a MonitorEVA based on: a) Vaccination status defined independently of number of days between vaccination and symptoms onset; b) use of all ILI data independent of the definition; c) stratification of VE according to time between onset and swab (< 3 and ≥3 days).

Effiziente Spurensicherung im Waffenlauf

Schüsse mit aufgesetzter Waffenmündung können biologische Antragungen im Waffenlauf hinterlassen, die durch Endoskopie optisch und durch PCR analytisch detektiert werden können. Im Rahmen des vom SNF geförderten Projektes wurden verschiedene Verfahren der Spurensicherung überprüft. Mit Vlies abgedeckte Farbpads (2 ml Acrylfarbe, 2 ml Humanblut, 1 ml Bariumsulfat) wurden in 12 cm grosse Gelatinewürfel integriert. Diese wurden mit aufgesetzter Waffenmündung mit verschiedenen Pistolen im Kaliber 7.65 mm Browning beschossen und die Waffenläufe endoskopiert. Die Spurensicherung erfolgte getrennt für den vorderen und den hinteren Laufabschnitt und wurde in mehreren Testreihen mit folgenden Mitteln durchgeführt: Wattetupfer (Holzstäbchen, Applimed), Forensic Swab (Sarstedt), COPAN FLOQSwab™, DNA-freie VFG-Filz-Stopfen. Anschliessend erfolgte eine endoskopische Kontrolle. Der DNA-Gehalt wurde mittels qPCR ermittelt. Grundsätzlich waren alle Verfahren für die Spurensicherung geeignet und erzielten eine gute Spurenausbeute. Wesentliche Unterschiede ergaben sich in der Handhabung. Holzstäbchen waren lang genug, jedoch rigide, was bisweilen zu ihrem Abbruch führte, andererseits aber guten Abrieb ergab. Der Forensic Swab war mit seinem flexiblen Kunststoffstiel leichter von hinten im Waffenlauf anzuwenden, was ihn auch für schwer zugängliche Stellen empfiehlt. Am einfachsten war eine gründliche Spurensicherung mit den FLOQs zu erreichen. Durch deren Nylonbürste wurde eine gute Abriebleistung bei flexiblem Stiel erzielt. FLOQs mit verschiedener Länge ermöglichten eine kontrollierte Spurennahme insbesondere im hinteren Teil des Waffenlaufes. FLOQs waren auch im molekular-biologischen Labor leicht zu verarbeiten. Mit den DNA-frei gemachten, kalibrierten VFG-Stopfen liessen sich restliche Spuren in teils erheblicher Konzentration mobilisieren. Allerdings sind Handhabung und Weiterverarbeitung im DNA-Labor wesentlich aufwändiger.

Transactions of the sixth International congress on tuberculosis. Washington, September 28 to October 5, 1908.

v. 1, pt. 1. Proceedings of Section I: Pathology and bacteriology. Proceedings of joint session of Sections I and II: Opsonic index. Conjunctival and cutaneous tuberculin reactions. Serum diagnosis.--v. 1, pt. 2 Proceedings of Section II: Clinical study and therapy of tuberculosis, sanatoria, hospitals, and dispensaries.--v. 2. Proceedings of Section III: Surgery and orthopedics. Proceedings of Section IV: Tuberculosis in infancy.--v. 3. Proceedings of Section V: Hygienic, social, industrial, and economic aspects of tuberculosis.--v. 4, pt. 1. Proceedings of Section VI: State and municipal control of tuberculosis.--v. 4, pt. 2. Proceedings of Section VII: Tuberculosis in animals and its relations to man.--v. 5. The opening and closing ceremonies. Report of the Secretary-General. Officers, committees, and members.--[v. 6] A series of public lectures Specially prepared for the sixth International congress on tuberculosis, by A. Calmette...Louis Landouzy...[and others] Ed. by the secretary-general, and printed as a supplement to the Transactions of the Congress.

Host and Environmental Correlates of Multi-Drug Resistance in Kenyan Children with Acute Bacterial Diarrhea

Thesis (Master's)--University of Washington, 2016-06

The treatment of pterygium

The treatment of pterygium is still quite controversial, with various treatments being advocated in the scientific literature. Unfortunately, there are very few well-conducted controlled clinical trials of treatments. However, years of anecdotal and noncontrolled studies have confirmed that some methods, such as bare scleral closure, are no longer acceptable in the treatment of pterygium and that other methods are likely to be more useful. In the future it will be important to develop a grading system, and surgeons will need to be conservative in the treatment of pterygium until such time as a single treatment provides a lower recurrence rate and complication rate.

Establishment and characterization of a new epithelial cell line, KC-1, from koala (Phascolarctos cinereus) conjunctiva

A novel, untransformed koala cell line (KC-1) was established by culturing koala conjunctival tissue in growth medium, which has permitted the study of the cell biology of this unique system. After the establishment of the KC-1 cell line, the cells were characterized by light microscopy, doubling time, and Western blot analysis. Light microscopy revealed that the cells have an epithelial morphology. Doubling times were significantly different (P < 0.015) depending on fetal calf serum (FCS) concentration (16.5 h in 10% FCS and 26.5 h in 2% FCS). Cells constricted while in suspension but were shown to attach to the coverslip (or flask) and flatten rapidly, less than 1 h after seeding. To confirm the epithelial nature of the cells, protein was extracted and Western blot analysis was performed. Subsequent probing with primary and secondary antibodies (monoclonal anticytokeratin clone C-11 IgG1 and anti-mouse IgG) revealed two bands at 45 and 52 kDa (compared against a protein molecular weight marker) that correspond to primary type I keratin and major type II keratin, respectively, expressed in simple epithelial cells. The koala cell line was adapted to grow continuously in Dulbecco modified Eagle medium containing 10% FCS for at least 30 passages. This unique cell line is an ideal toot for further investigation on koala cell biology and cytogenetics and for exploration of the pathophysiological mechanism of eye infections caused by different pathogens in koalas.