Two Regulators of Vibrio parahaemolyticus Play Important Roles in Enterotoxicity by Controlling the Expression of Genes in the Vp-PAI Region

Vibrio parahaemolyticus is an important pathogen causing food-borne disease worldwide. An 80-kb pathogenicity island (Vp-PAI), which contains two tdh (thermostable direct hemolysin) genes and a set of genes for the type III secretion system (T3SS2), is closely related to the pathogenicity of this bacterium. However, the regulatory mechanisms of Vp-PAI's gene expression are poorly understood. Here we report that two novel ToxR-like transcriptional regulatory proteins (VtrA and VtrB) regulate the expression of the genes encoded within the Vp-PAI region, including those for TDH and T3SS2-related proteins. Expression of vtrB was under control of the VtrA, as vector-expressed vtrB was able to recover a functional protein secretory capacity for T3SS2, independent of VtrA. Moreover, these regulatory proteins were essential for T3SS2-dependent biological activities, such as in vitro cytotoxicity and in vivo enterotoxicity. Enterotoxic activities of vtrA and/or vtrB deletion strains derived from the wild-type strain were almost absent, showing fluid accumulation similar to non-infected control. Whole genome transcriptional profiling of vtrA or vtrB deletion strains revealed that the expression levels of over 60 genes were downregulated significantly in these deletion mutant strains and that such genes were almost exclusively located in the Vp-PAI region. These results strongly suggest that VtrA and VtrB are master regulators for virulence gene expression in the Vp-PAI and play critical roles in the pathogenicity of this bacterium

Diagnóstico laboratorial da infecção pela Chlamydia trachomatis: vantagens e desvantagens das técnicas

O objetivo deste artigo é revisar e comentar as vantagens e desvantagens dos diferentes tipos de testes de detecção de Chlamydia trachomatis na rotina de laboratórios clínicos, com ênfase nas técnicas de amplificação. A Chlamydia trachomatis é considerada a bactéria sexualmente transmissível mais freqüente em países desenvolvidos e de grande impacto no sistema reprodutivo das mulheres. É o agente causador de doenças do trato urogenital, linfogranuloma venéreo (LGV), tracoma, conjuntivite de inclusão e pneumonia no recém-nascido. Um dos fatores de risco para a infecção é a prática sexual entre adolescentes. A recorrência das infecções é comum. Episódios sucessivos de infecção aumentam o risco de desenvolver seqüelas e a chance de contrair a infecção pelo vírus da imunodeficiência humana. O diagnóstico da infecção pela Chlamydia trachomatis ainda é crítico, devido à freqüência de infecções assintomáticas. As técnicas de amplificação de ácidos nucléicos permitem utilizar urina para a detecção da clamídia, simplificando a coleta. Apresentam maior sensibilidade do que a cultura e do que os testes mais utilizados, como a imunofluorescência direta e o enzimaimunoensaio. A cultura celular, utilizada como padrão- ouro, tem especificidade de 100% e sensibilidade de 70% a 85%. De acordo com o Centers for Disease Control (CDC), um diagnóstico é considerado definitivo quando positivo em cultura ou em pelo menos dois testes não-culturais distintos. Os testes de amplificação são mais dispendiosos do que os demais testes não-culturais, mas de menor custo que a cultura.<br>The purpose of this article is to review and comment the advantages and disadvantages of Chlamydia trachomatis tests in clinical laboratories routine, emphasizing the techniques of amplification. Chlamydia trachomatis is considered the most frequent sexually transmitted bacterium in developed countries and it has an important impact on reproductive female tract. It causes diseases on urogenital tract, venereal limphogranuloma, trachoma, conjunctivitis and newborn pnemonia. One of the main risks of infection is the sexual practice among adolescents. Repetitive episodes of infection raise the risk of developing sequel as well as the chance of getting the infection with the human immunodeficiency virus. Its hard to diagnose the infection caused by Chlamydia trachomatis because most of times there are no symptoms. Nucleic acid amplification techniques allow us to use urine to detect chlamydia. It simplifies the sampling and offers greater accuracy than bacteria culture. It is even better than tests such as direct fluorescent antibodies and immunoassay. Cellular culture as reference techniques results in 100% of specificity and 70 to 85% of sensitivity. According to the Centers for Disease Control (CDC) is considered definitive when positive culture or, at least, two non culture distint tests. Amplification tests are more expensive than other tests, but cheaper than culture