To investigate sodium-dependent growth, ‘sodium-free M9’ was prep

To investigate sodium-dependent growth, ‘sodium-free M9’ was prepared by replacing PLX4032 order all sodium salts in M9 (this is around 50 mM Na+ in normal M9) with their potassium equivalents and replacing agar with 0.8% agarose; the sodium-free M9 medium still contained approximately 50 μM sodium from the Amp used for selection. M9 agarose plates required longer incubation

times (up to 4 days) for single colonies to grow. All experiments involving the WT and the ΔnanT strains used cells transformed with the empty vector, i.e. pWKS30. The presence of the vector did not affect the growth phenotypes of either strain (not shown). Starter cultures were prepared as described for the growth experiments, except that o/n growths were carried out in M9 Amp supplemented with 2 mg mL−1 glucose and 1 mM IPTG. Overnight cultures were diluted to an OD650 nm of 0.1 in the

same medium and allowed to grow at 37 °C until they reached an OD650 nm of 0.5, when they were harvested, washed four times in M9, resuspended in the same buffer at a final OD650 nm of 3 and stored PD0332991 clinical trial on ice till use. For Neu5Ac uptake assays, cells were diluted 10-fold in M9 prewarmed at 37 °C and allowed to acclimatize for 2 min, with stirring before initiating the assays by adding of varying amounts of [14C]-Neu5Ac (Sigma) appropriately diluted with unlabelled Neu5Ac. The uptake assay and total protein quantification were then performed as described in Severi et al. (2008), except that 200 μL of cell suspensions were immobilized instead of 400 μL. [14C]-Neu5Ac was normally used at a final concentration of 0.5–2 μM and isotopically diluted (up to 100 ×) with unlabelled Neu5Ac when required. Ks and Vmax values were calculated by fitting the experimental data for uptake rates to a hyperbolic Michaelis–Menton equation using sigmaplot. To assay sodium-dependent Neu5Ac uptake, cells were prepared as for a standard uptake assay, except that sodium-free M9 (see the previous section) was used as both washing and

assay buffers. Salts, i.e. NaCl, KCl and LiCl, were added at a final Resveratrol concentration of 100 mM during the acclimitization phase. The assay was performed as described above with a concentration of 100 μM total Neu5Ac. Cold chase experiments were performed as described in Mulligan et al. (2009), using SEVY1 cells transformed with the appropriate plasmids. To assess the suitability of an E. coliΔnanT strain for the functional characterization of hypothetical Neu5Ac transporters, we first examined cells expressing either nanT itself or the known siaPQM TRAP transporter genes from H. influenzae cloned into a low-copy-number vector under the control of an IPTG-inducible promoter.

Capsule enlargement in C neoformans requires extracellular deliv

Capsule enlargement in C. neoformans requires extracellular deliverance of GXM, which is further incorporated into the fungal cell surface to promote distal capsular growth (reviewed in Zaragoza et al., 2009). The subsequent self-aggregation of polysaccharide molecules occurs by mechanisms that putatively require divalent cations, such as calcium II (Nimrichter et al., 2007). The inhibitory activity of microplusin on capsular

enlargement could be due to the interference with aggregation of the building blocks through metal chelation, thereby affecting the correct polysaccharide capsule assembly. However, based on our mass spectrometry analysis, microplusin does not bind calcium II (Silva et al., 2009). Thus, its effect on capsule enlargement GDC-0980 ic50 Selleckchem Dasatinib most likely results from inhibition of one or more metabolic processes dependent on enzymes that requires copper as a cofactor. Notably, the Δvph1 mutant also had aberrant capsular production (Li & Kaplan,

1998; Erickson et al., 2001). In conclusion, microplusin showed a noteworthy fungistatic activity in vitro against C. neoformans. We demonstrate that this effect may be related to its inhibitory effect on the classical respiratory pathway of C. neoformans. Microplusin also affected the two most important virulence factors of this mycopathogen: the melanization process and the formation of a polysaccharide capsule. These findings are particularly relevant for determining the utility of copper-chelator compounds, like microplusin as a therapeutic for cryptococcosis.

However, studies in vivo are Oxymatrine crucial to corroborate the efficiency of this peptide or other metal chelators for combating C. neoformans. S.D. is supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); M.L.R. is supported by CNPq, Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); J.D.N. is supported in part by RO1 AI52733 and by the Einstein-Montefiore CFAR (NIH AI-51519). L.R.M. gratefully acknowledges support from Long Island University. We are grateful to Susana P. Lima for technical assistance and Cassiano Pereira for figure preparation. “
“Adherent growth of Pseudomonas putida KT2440 with and without the TOL plasmid (pWWO) at the solid–liquid and air–liquid interface was examined. We compared biofilm formation on glass in flow cells, and assayed pellicle (air–liquid interface biofilm) formation in stagnant liquid cultures by confocal laser scanning microscopy. The TOL-carrying strains formed pellicles and thick biofilms, whereas the same strains without the plasmid displayed little adherent growth. Microscopy using fluorescent nucleic acid-specific stains revealed differences in the production of extracellular polymeric substances: TOL carriage leads to more extracellular DNA (eDNA) in pellicles and biofilms.

“To evaluate the disease activity and current pharmacologi

“To evaluate the disease activity and current pharmacological interventions used to achieve remission in rheumatoid arthritis (RA) patients in Australia. Rheumatoid arthritis patients treated in participating Australian clinics were included in the study. Patient demographics, disease onset, medications and disease measures were analyzed. Data, de-identified to the patient, clinic and clinician were captured using an electronic selleck clinical management program. The disease activity score

(DAS28) was used to classify patients into the disease activity states of remission, low disease activity (LDA), moderate disease activity (MDA) and high disease activity (HDA). Choice of therapy was at the discretion of the treating clinician. A total of 5686 patients, 72.9% female, 26.9% male, with mean age 61.1 (SD 13.6) years

and mean disease duration of 11.5 (SD 10.5) HKI-272 order years were analyzed. DAS28 ESR (erythrocyte sedimentation rate) scores were recorded for 2973 patients, with 41.6% in remission, 18.6% LDA, 31.6% MDA and 8.2% HDA. Of those in remission, 17% received a biological disease modifying anti-rheumatic drug (bDMARD), 73% methotrexate (MTX), 19% leflunomide (LEF) and 28% prednisolone. Of the patients with MDA, 20% received a bDMARD, 76% MTX, 24% LEF and 39% prednisolone. Of the patients in HDA, 27% received a bDMARD, 78% MTX, 31% LEF and 60% with prednisolone. Cross-sectional assessment of this large cohort of Australian RA patients found a large proportion remain in moderate or high disease activity; suggesting a considerable evidence–practice gap. Improvement in disease control in this group may reduce future health burdens. “
“To describe the clinical manifestations, disease activity and organ damage in Korean patients with systemic lupus erythematosus (SLE).

American College of Rheumatology (ACR) criteria, SLE Disease Activity Index (SLEDAI), and Systemic Lupus International Collaborating Clinics/ACR damage index (SDI) were assessed in patients with SLE from 1998 to 2012. A total of 996 SLE patients were analyzed. The common accrual of ACR criteria included: immunologic (93%), hematologic (93%), arthritic (66%) and nephritic (50%). In the inception cohort over 10 years of follow-up Bumetanide (n = 120), the number of ACR criteria increased significantly (5.0 ± 1.2 to 5.7 ± 1.3), and nephritis, serositis and neuropsychiatric symptoms tended to increase continuously over time. SLEDAI-2K decreased significantly (5.6 ± 3.4 to 4.1 ± 1.2), but the percentage of patients with SLEDAI scores ≥ 12 did not decrease over time. The common organ damages were musculoskeletal (14.9%) and renal (11.1%). The mean SDI score increased significantly (0.4 ± 0.8 to 1.1 ± 1.6) and renal damage had two peaks in 1 and 6–10 years, musculoskeletal and neuropsychiatric damage were predominant from 1 to 5 years, and ophthalmic damage increased sharply over 10 years.

Posterior probability (PP) values were subsequently calculated S

Posterior probability (PP) values were subsequently calculated. Stabilization of model parameters (burn-in) occurred around 2 400 000 and 800 000 generations for 16S rRNA and surface-encoding genes, respectively. Every 100th tree after stabilization (burn-in) Hydroxychloroquine in vivo was sampled to calculate a 50% majority-rule

consensus tree. All trees were constructed using the program figtree v1.3.1 ( dnasp (Librado & Rozas, 2009) was used to calculate synonymous (dS) and nonsynonymous (dN) rates and two common measures of nucleotide variation, π and θW, for determining ompA intraspecies variation within Glossina. Neutrality tests were also performed in dnasp. The McDonald–Krietman test and neutrality index

(NI) were calculated by comparing the ratio of dS to dN mutations within either individual Glossina species for ompA, or among Glossina isolates for ompC, and an E. coli outgroup. The outgroup was composed of ecologically diverse E. coli representatives NC_000913, selleck inhibitor NC_008253, and NC_002655. These adaptive evolution tests have been shown to be most powerful when taxa are closely related (Clark et al., 2003). We chose E. coli as our representative outgroup because it is a close relative of Sodalis, and has a wide representation of publicly available genome strains. The

nucleotide sequences determined in this study have been deposited in the NCBI GenBank database under accession numbers HM626140–HM626149 and HQ914651-HQ914697. To examine the evolutionary relationships of the newly identified Sodalis-like symbionts, we constructed phylogenetic trees based on 16S rRNA gene sequences. Bayesian analysis supports the monophyly of Gammaproteobacteria symbionts learn more isolated from diverse insect orders (i.e. Diptera, Coleoptera, Hemiptera, and Phthiraptera) (Fig. 1). In general, there is a tight clustering of symbionts with respective insect host Order. Our Bayesian analysis also suggests the closer relationship of hippoboscid symbionts to weevil and pigeon louse symbionts, rather than to Sodalis, despite a common ancestry of their respective hosts within the Hippoboscoidea (Petersen et al., 2007), thus further substantiating a previous hypothesis of independent symbiont acquisition events by these hosts (Novakova & Hyspa, 2007). However, there is only moderate Bayesian support for this relationship (PP=77, data not shown) that is further decreased (PP=51) when symbionts of the recently reported chestnut weevil Curculio sikkimensis (Toju et al., 2010) and the stinkbug Cantao ocellatus (Kaiwa et al., 2010) are included in the analyses.

7,8 Since our patient immigrated to Germany only 2 years before i

7,8 Since our patient immigrated to Germany only 2 years before initial diagnosis, and has never visited southern Germany’s AE endemic areas, it is suggestive that he acquired the disease in Siberia, a highly endemic region. Surveillance systems are not standardized in most endemic countries. In some countries surveillance does not exist at all, therefore incidence rates might be strongly underestimated. The annual global incidence is estimated to be

approximately 18,235 cases (0.26/100,000), of which 16,629 [(91%); 1.24/100,000] have been described in learn more China, and 1606 cases outside China.9,10 Globalization and increased immigration of people from highly endemic to non-endemic areas could potentially raise the number of cases in non-endemic areas.11 Epigenetic inhibitor However, the exposure risk of the usual, short-term traveler to acquire AE is minimal; no cases have been reported so far. Cerebral AE as a differential diagnosis needs to be considered in patients presenting

with neurological symptoms, cerebral lesions, eosinophilia of unknown origin, and who live in or are returning from endemic areas. In endemic areas, regular serological testing and imaging procedures would be important tools for early detection. In general, positive serology does not necessarily confirm diagnosis as antibody titers can also be interpreted as serum residuum titers, ie, in our

patient from hepatic AE. Serology is positive in up to 80% of cases; cross reaction with other helminths is possible. However, recent advanced serology using recombinant antigens such as RecEm18 appears to detect more than 95% of active AE with almost no cross reaction with non-echinococcus diseases.12 Histopathological diagnosis from Teicoplanin tissue specimen is the gold standard but not available universally. In addition to that, it is especially difficult to obtain from the brain. A polymerase chain reaction has been established in specialized laboratories. Molecular diagnostic from tissue specimen might be helpful in selected cases. The treatment of cerebral AE is often difficult: surgical removal, followed by at least 2 years, sometimes life-long chemotherapy is the standard therapy. Treatment with benzimidazoles is the preferred option.4,13 In inoperable disease, chemotherapy with anthelmintic medication is the only treatment shown to be potentially effective, but is usually palliative.13 Because of its better resorption ABZ, compared to mebendazole, is the treatment of choice. Serum levels of ABZ and its active metabolite ABZ-sulfoxide should be monitored for dose adjustments and thus the prevention of side effects and disease progression. Failure to reach therapeutic drug levels or eradicate viable lesions remains a problem, as shown in our patient.

A high proportion of the earlier published cases of JE have been

A high proportion of the earlier published cases of JE have been in the United States and allied military personnel stationed in SEA regions. From 1945 to 1972, 131 cases of JE were reported in military personnel. In the years 1978 to 1992 and 1992 to 2008, 24 cases and 21 cases were reported, respectively. Rates of 0.1–2.1 per 10,000 per week have been observed in nonimmunized US military personnel in Asia.[7] JE vaccination is recommended for this group of travelers. JE infection has been reported in short-term travelers who Selleck Pirfenidone have traveled outside of the rainy season with minimal travel to rural

locations.[3, 8-10] This has raised concerns about the limits in our current understanding of the risk of JE infection in short-term travelers. Characterizing CX-5461 price the current risk of JE in general travelers and the uncertainty limits around this risk provides valuable information to travel medicine practitioners advising prospective travelers. In our cohort of predominantly short-term travelers, travel was more common in periods of the year where JE transmission is higher and whilst nearly half of the

travelers visited or stayed in a rural area overnight, only a small proportion of travel-days were spent on “outdoor” activities. The risk of JE infection is linked to outdoor exposure in the dusk or evening times in rural destinations where JE transmission occurs. In terms of adherence to pre-travel advice, most travelers utilized some form of mosquito preventive behavior, although consistency of use was not documented. Only a small proportion of travelers (9%) were vaccinated for JE, which probably reflects the current recommendations for JE vaccine, in that a majority were short-term travelers and not spending a great amount of time in rural areas. Low-level antibodies at baseline were noted in 2.8% of travelers with possibilities

of previous JE, given the presence of JE in Northern Australia, or other flavivirus vaccination or Baricitinib infection as possible explanations. A limitation of this study is the potential impact of a small sample size on the likelihood of observing an infrequent infection such as JE (clinical or subclinical) in travelers. A further limitation is the generalizability of the findings from a travel-clinic attendee cohort who may be different to general travelers. Data were also incomplete for the travelers who did not complete the study. Although unlikely, it is also possible that some seroconversions were missed given the timing of the second bleed (day 10). Several considerations relating to risk factors for infection, adverse effects and costs of vaccine, and individual personal preference regarding vaccination, need to be considered when discussing indications for or against vaccination. The threshold for JE vaccination is generally still based on historical risk-benefit considerations that may no longer be valid now as we have a safer vaccine.

Comparison of ClinSurv HIV with other ongoing clinical HIV cohort

Comparison of ClinSurv HIV with other ongoing clinical HIV cohort studies suggests that the ClinSurv HIV data might play a more important role in the future in complementing HIV research in European countries Doxorubicin price with concentrated HIV

epidemics [7–9]. Close collaboration with European HIV drug resistance networks [the Collaborative HIV and Anti-HIV Drug Resistance Network (CHAIN) and the Collaboration of Observational HIV Epidemiological Research Europe (COHERE)] is already ongoing or planned for the near future. After almost 10 years of data collection, the German national ClinSurv HIV cohort study has evolved to become a valuable and effective tool for clinical surveillance. Its database is stored and managed at the Unit for HIV/AIDS, STI and Blood-Borne Infections of the Department of Infectious Disease Epidemiology of the RKI in Berlin. It is hoped that this cohort study will make significant contributions to answering epidemiological and clinical research questions in the future in countries such as Germany with concentrated HIV

epidemics. ClinSurv HIV is interested in further national and international research co-operation in the area of clinical HIV epidemiology. Additional information about ClinSurv HIV is provided on the homepage of the RKI [25]. Pexidartinib in vitro Berlin: Charité, Universitätsmedizin Berlin, Campus Rudolph Virchow (Dr F Bergmann and Prof. Dr N Suttorp); Vivantes-Klinikum, Auguste-Viktoria-Krankenhaus (Priv.-Doz. Dr K Arasteh)*. Bochum: Ruhr Universität Bochum, St Joseph Hospital (Prof. Dr N Brockmeier)*. Bonn: Rheinische Friedrich-Wilhelm Universität Bonn (Prof. Dr J Rockstroh). Düsseldorf: Universitätsklinikum Düsseldorf (Dr S Reuter and Prof. Dr D Häusinger). Essen: Universitätsklinikum Essen, Klinik für Dermatologie (Dr S Esser)*. Hamburg: Institut für interdisziplinäre Infektiologie Dynein (ifi) (Prof. Dr A Plettenberg); Bernhard Nocht-Institut (Prof. Dr G-D Burchard)†; Universitätsklinikum Hamburg-Eppendorf (Priv.-Doz.

Dr J van Lunzen); Infektionsmedizinisches Centrum Hamburg (ICH), ICH Study Center (Dr K Schewe, Dr L Weitner, Dr A Adam, Dr H Gellermann, Dr S Fenske, Dr T Buhk, Prof. Dr H-J Stellbrink and Priv.-Doz. Dr C Hoffmann). Hannover: Medizinische Hochschule Hannover (Prof. Dr M Stoll and Prof. Dr RE Schmidt). Kiel: Universitätsklinikum Kiel (Prof. Dr H Horst). Köln: Universität zu Köln (Dr T Kümmerle and Prof. Dr G Fätkenheuer). München: Ludwig-Maximilians-Universität München (Prof. Dr J Bogner). Rostock: Universitätsklinikum Rostock (Dr C Fritsche and Prof. Dr EC Reisinger). All persons listed are members of the ClinSurv HIV Study Group. *The inclusion of data from these three treatment centres in the ClinSurv HIV cohort is currently in preparation. Since 2002 this centre has not actively contributed patient data; however, previously reported case events remain within the observational database. The authors are grateful to all collaborative treatment centres listed in the Appendix.

Since most sites reported that patients were CD4 tested at least

Since most sites reported that patients were CD4 tested at least annually, CD4 monitoring was classified

into two categories: at least three times and fewer than three times per year. The two exceptions monitored patients at least annually when resources were available to do so. Clinical disease progression was determined as a new diagnosis of an AIDS-defining illness (CDC category C) or death from any cause. Patient follow-up commenced at HAART initiation and ended at date of death, AIDS-defining illness or most recent contact, whichever was the earliest. Surrogate endpoints were HIV RNA viral suppression (<400 copies/mL) and change Epacadostat in CD4 cell count from baseline at 12 months post-HAART. Surrogate

marker values closest to the target date were selected from windows of 9–15 months. Patients contributing data to each analysis are shown in Fig. 1. For eligible patients, baseline comparisons by country income (χ2, Fisher’s exact or Cochrane – Armitage test for trend) were performed as appropriate. Determinants of 12-month HIV RNA suppression and change in CD4 cell count were assessed via logistic regression and linear regression, respectively. Proportional hazards models were used to evaluate predictors of time to progression to new AIDS-defining illness or death. Analyses were based on an intention to continue treatment approach in that we did Proteasome inhibitor not take into account regimen changes, interruptions or failure post-HAART. Forward stepwise techniques were used to determine the best fitting models. To identify significant variables Thymidine kinase and important confounders, binary covariate P-values and multi-categorical parameter P-values (from tests for trend/heterogeneity) of <0.2, in univariate analyses, were considered for inclusion in multivariate models. Final multivariate models consisted of covariates remaining significant at the 0.05 level. For each endpoint, a base predictive patient model was determined from significant patient covariates. Then, because of our a priori interest in the role of site resourcing

on outcomes, individual estimates of country income and reported frequencies of VL and CD4 testing were assessed for statistical significance after adjustment for the base patient model. Analyses were performed using sas software version 9.1.3 (SAS Institute Inc., Cary, NC, USA) and stata software version 8.2 (STATA Corp., College Station, TX, USA). Of 3346 patients recruited to TAHOD, 2333 (69.7%) fulfilled the inclusion criteria. Of these, 79% had at least 6 months of retrospective data available and 13% were mono- or dual-ARV experienced. Patient demographics, clinical parameters and prescribed HAART regimen are summarized in Table 1. One hundred and seventy-six of the mono- and dual-experienced patients recycled one or two previously used ARVs in the HAART regimen.

Visceral leishmaniasis in HIV-seropositive individuals usually oc

Visceral leishmaniasis in HIV-seropositive individuals usually occurs in those with CD4 counts below 200 cells/μL [29]. Leishmania cause three types of disease: Visceral (kala azar), which usually presents with systemic features of fever and weight loss along with hepatosplenomegaly (with splenic enlargement most prominent), with or without bone marrow involvement; Most reported cases of HIV/Leishmania co-infection in Europe are of visceral leishmaniasis Belnacasan order [30]. Cases may be associated with a history of intravenous

drug use [31]. Visceral leishmaniasis usually, but not always, presents in the same way as it does in HIV-seronegative people; the systemic features may be mistaken for other opportunistic infections. Cutaneous leishmaniasis may present as it does in immunocompetent individuals with a papule that progresses to a Lumacaftor in vivo chronic ulcer, but a wide range of atypical skin lesions may occur, and may be mistaken for Kaposi’s sarcoma or bacillary angiomatosis. Isolated mucocutaneous leishmaniasis in association with HIV infection appears to be very rare in Europe, probably as L. infantum, which causes most visceral

leishmaniasis in Europe, rarely causes mucosal lesions. However, any patient with a suspected leishmanial lesion on the face should be seen urgently by a specialist. Mucocutaneous leishmaniasis may be seen in cases acquired in Central or South America where the infecting species have greater tropism for mucous membranes. Diagnosis of leishmaniasis mafosfamide requires parasitological or histological confirmation (category III recommendation). Diagnosis depends on parasitological or histological demonstration of Leishmania. Parasitological diagnosis is most useful because identification of Leishmania species may guide appropriate treatment. In the context of HIV, standard diagnostic tests may be less sensitive and expert advice should be sought (category

IV). Visceral leishmaniasis. Parasitological diagnosis may be made by microscopy, culture or PCR. Appropriate specimens include [30,32,33]: Splenic aspirate: this has the highest sensitivity, but should only be performed by a practitioner trained in the technique; It is strongly recommended to liaise with the local tropical disease and parasitology service before taking specimens. Some transport media (e.g. those with antifungal agents) may inhibit leishmania culture so specimen transport should be discussed with the laboratory. Histological diagnosis may be made on biopsy of bone marrow, lymph node, liver, skin or other tissue. Serological tests include the direct agglutination test and ELISA to detect antibodies to recombinant K39 antigen (rK39). The sensitivity of both may be reduced in HIV/Leishmania coinfection [32] due to low levels of antibody in HIV-seropositive individuals [34]. Cutaneous leishmaniasis. Parasitological or histological diagnosis (preferably both) may be made from a skin biopsy [32].

parahaemolyticus possesses a full set of the exsACDE regulatory s

parahaemolyticus possesses a full set of the exsACDE regulatory system, which is similar to that of P. aeruginosa and which regulates T3SS1-related gene expression. H-NS is a major component of the bacterial nucleoid and plays a crucial role in global gene regulation in enteric bacteria (Varshavsky et al., 1977; Hulton et al., 1990). H-NS affects the expression of many unrelated genes and several virulence genes in Salmonella learn more enterica serovar Typhimurium, Shigella sp. and Vibrio cholerae (Maurelli & Sansonetti, 1988; Hulton et al., 1990; Tobe et al., 1993; Harrison et al., 1994; O’Byrne & Dorman, 1994;

Nye et al., 2000). Therefore, we examined the possibility that T3SS1 genes are part of the H-NS regulon. As shown in Fig. 3a, the production of VscC1 and VepA proteins in a Δhns strain was considerably increased in both the bacterial pellet and the supernatant compared with that of the WT. A ΔhnsΔexsA mutant strain did not exhibit

increased production of these proteins (Fig. 3b), suggesting that exsA is necessary for overproduction of T3SS1-related proteins via hns gene deletion. We next examined the possibility that H-NS represses exsA expression using an exsA–lacZ transcriptional fusion reporter (Fig. 3c). Transcription of exsA–lacZ was dramatically increased in the hns deletion strain compared with that in the WT. The increase in exsA–lacZ transcription in the hns deletion strain was suppressed by Selleck 5-Fluoracil in trans complementation with the Selleck ZD1839 hns gene. These results

indicate that H-NS represses T3SS1-related gene expression by suppressing exsA gene expression. In summary, we identified VP1701 and VP1702 of V. parahaemolyticus as functional orthologues of P. aeruginosa ExsC and ExsE, respectively. As VP1701 has sequence similarity with its counterpart, it was not difficult to predict its function. Indeed, the production of T3SS1-related proteins was repressed in a Δvp1701 mutant and derepressed by complementation of the vp1701 gene (Fig. 1b and c). Unlike ExsA (VP1699), ExsD (VP1698) and ExsC (VP1701), sequence annotation of the T3SS1 region on the genome of V. parahaemolyticus did not reveal any CDSs predicted to encode the homologue of P. aeruginosa ExsE. However, we found that one hypothetical CDS (VP1702) was encoded next to the vp1701 (exsC) gene. Deletion of the vp1702 gene deregulated the production of T3SS1-related proteins. Furthermore, VP1702 itself was a substrate for the T3SS1 secretion system. These properties of VP1702 of V. parahaemolyticus conform with those of its counterpart in P. aeruginosa. In P. aeruginosa, the coupling of transcription to secretion is mediated by three interacting proteins (ExsC, ExsE and ExsD) that regulate ExsA transcription activity (Yahr & Wolfgang, 2006). Although it is still unknown whether ExsC, ExsE and ExsD of V.