MVL and TB wrote the paper. MVL, TB, ZBH, GK and NO interpreted the data. TB, ZBH, GK, RS, NO, JG, CP and CSL were responsible for critical revision of the paper and for important intellectual content. TB, RS, GK, NO, JG, CP and CSL carried out data collection. Financial support: MVL has received grants from Region Hovedstaden and Preben og Anna Simonsens Fond. The Danish HIV Cohort Study has received financial support from the University of

Copenhagen, Rigshospitalet and the NOVO Nordisk Foundation. Conflicts of interest: NO has received research funding from Roche, Bristol-Myers Squibb, Merck Sharp & Dohme, GlaxoSmithKline, Abbott, Boehringer Ingelheim, Janssen-Cilag and Swedish Orphan. TB has received honoraria and consultancy fees from Bristol-Myers Squibb, GlaxoSmithKline, and Statens Serum Institut, Copenhagen. Dabrafenib mouse RS has received honoraria and consultancy fees from Pfizer, is on the advisory board for Leo Pharma, Novartis and Targenta selleck and has been a speaker at Novartis symposiums. All other authors have no conflicts of interest to declare. “
“The aim of this study was to examine Emergency Department (ED) utilization and clinical and sociodemographic correlates of ED use among HIV-infected patients. During 2003, 951

patients participated in face-to-face interviews at 14 HIV clinics in the HIV Research Network. Respondents reported the number of ED visits in the preceding 6 months. Using logistic regression, we identified factors associated with visiting the ED in the last 6 months and admission to the hospital from the ED. Thirty-two per cent of respondents reported at least one ED visit in the last 6 months. In multivariate analysis, any ED use was associated with Medicaid insurance, high levels of pain (the third or fourth quartile), more than seven PRKD3 primary care visits in the last 6 months, current or former illicit drug use, social alcohol use and female gender. Of those who used ED services, 39% reported at least one admission to the hospital. Patients with pain in the highest quartile reported increased admission rates from the ED

as did those who made six or seven primary care visits, or more than seven primary care visits vs. three or fewer. The likelihood of visiting the ED has not diminished since the advent of highly active antiretroviral therapy (HAART). More ED visits are to treat illnesses not related to HIV or injuries than to treat direct sequelae of HIV infection. With the growing prevalence of people living with HIV infection, the numbers of HIV-infected patients visiting the ED may increase, and ED providers need to understand potential complications produced by HIV disease. HIV-infected patients are more intensive users of the healthcare system than the general population [1,2]. Studies early in the HIV epidemic demonstrated that this population had a higher-than-average rate of Emergency Department (ED) use compared with the general US population [3].

Only two patients in the combined NVP arm and two patients in the ATZ/r arm of the study experienced cardiac disorders of division of acquired immunodeficiency syndrome (DAIDS) grade 3 or 4. In the combined NVP arm, one patient experienced angina pectoris and one patient Selumetinib in vivo experienced myopericarditis. In the ATZ/r arm, one patient experienced MI, and another experienced cardiac failure. Primary data from the ARTEN study confirm that the favourable virological and immunological responses to NVP combined with TDF/FTC are maintained through 48 weeks of treatment and are noninferior

to those of ATZ/r [in combination with the same dual nucleoside reverse transcriptase inhibitor (NRTI) backbone] with a similar safety profile [23]. The data presented here also suggest a more favourable lipid profile with NVP than with ATZ/r when combined with TDF/FTC. There are many risk factors for CVD. Known factors include smoking, being overweight, lack of exercise, insulin resistance, elevated waist circumference, hypertension, elevated LDL-c, elevated triglycerides and low HDL-c. For HIV-infected patients receiving treatment with ARVs, the risk of CVD may be significantly greater than in the general population [27]. Increased levels of TG, TC and LDL-c, reduced levels of HDL-c, unfavourable changes in the TC:HDL-c ratio and lipodystrophy are common side effects in patients receiving certain ARV drugs

[1–4]. The cardiac disorders of DAIDS grade 3 or 4 reported in four patients in the ARTEN study (two in each arm) probably relate to pre-existing cardiovascular Mirabegron risk factors, although a role of antiretroviral therapy (ART) cannot be ruled selleck screening library out. With respect to serum lipid levels, traditionally LDL-c is recognized as the primary target of cholesterol-lowering therapy. However, full evaluation of lipid-related risk (i.e. TC, HDL-c, the TC:HDL-c ratio and TG levels) should

also be considered, as these measures play an important role as markers of cardiovascular risk [28]. Although ATZ/r use was associated with markedly lower LDL-c increases compared with NVP, LDL-c is known to be an incomplete measure of atherogenic lipoproteins because very low-density lipoprotein (VLDL) remnants are also likely to contribute to coronary heart disease [29]. In contrast, ApoB measurement includes all atherogenic lipoproteins, with each VLDL and LDL particle having one molecule of ApoB, making ApoB a more reliable measure of the concentration of proatherogenic particles [30]. The Apolipoprotein-related Mortality Risk (AMORIS) study showed that elevated ApoB levels were strongly related to increased cardiovascular risk and were also a stronger marker of cardiovascular risk than LDL-c [31]. In the current study, NVP-containing regimens showed no difference in ApoB, significantly greater increases in HDL-c and ApoA1, and an improved ApoB:ApoA1 ratio over 48 weeks compared with the ATZ/r regimen.

The reactions were performed as per the manufacturer’s instructions www.selleckchem.com/HIF.html (New England Biolabs, UK). DNA fragments were electrophoresed on 1% agarose gel at 5 V cm−1, in 1× TAE buffer (40 mM Tris, 40 mM acetate, 2 mM EDTA, pH 8.0). φ Lambda HindIII digest was used as DNA molecular weight marker. Gels were stained with ethidium bromide and photographed. Pulsed-field gel electrophoresis (PFGE) of bacteriophage DNA was carried out using the protocols described earlier (Carlson, 2005). Agarose plugs were prepared by mixing 1 mL of bacteriophage concentrate with 1 mL of 1.6% PFGE-grade agarose (Bio-Rad, Hercules, CA) and allowed to solidify at room temperature. These agarose

plugs were incubated in EDTA sarcosine proteinase K (ESP) (0.5 M EDTA, pH 9,

1% N-laurylsarcosine, and mg mL−1 proteinase K) overnight at 50 °C to digest the bacteriophage coat proteins and then washed thrice with TE (10 mM Tris-HCl, 0.1 mM EDTA, pH 7.5). The plugs were then treated with RNase A (μg mL−1) by incubating at 37 °C for 30 min. Restriction digestion was performed with various enzymes such as AluI, BamHI, BgII, DraI, HindIII, HaeII, KpnI, NcoI, NotI, PstI, XbaI, and ScaI following manufacturer’s instructions. The plugs were cut to 2-mm slices, placed in 1× restriction buffer, and incubated for 10 min in a 37 °C water bath. The buffer was removed, and fresh restriction buffer selleck containing 10 U of enzyme was added and incubated at 37 °C in water bath for 4 h. PFGE was carried out in 1% agarose gels in a BioRad CHEF DR-III PFGE

system (Bio-Rad), at 120° angle and 6 V cm−1, using ramped pulse times from 1 to 12 s for 6 h in 0.5× TBE (45 mM Tris, 45 mM borate, 1 mM EDTA pH 8.0) at 14 °C. Low-range PFGE marker was used as molecular weight size standard. Genome size was estimated by adding the length of each DNA fragment in the PFGE profile of ScaI and XbaI separately. The REA and PFGE patterns were captured using the Quantity one electrophoresis analysis system (Bio-Rad). Gel images were digitally normalized Protein kinase N1 to a single DNA marker to reduce gel-to-gel restriction pattern variability, and cluster analysis was carried out using molecular analyst software – Fingerprinting II (Version 3.0; Bio-Rad) by unweighted pair group method with arithmetic mean. Ability of the phages to transduce genetic elements was demonstrated by the transduction of the plasmid pHSG396 (Takara Bio Inc., Shiga, Japan), which possesses two selective phenotypic markers, β-galactosidase and chloramphenicol resistance. An isolate of V. harveyi (Vh57) susceptible to all four phages was transformed by CaCl2 treatment (Sambrook et al., 1989). Transformants carrying the plasmid were grown in 10 mL PYSS broth supplemented with 50 μg mL−1 chloramphenicol at 30 °C. This broth was suitably diluted with sterile PYSS to obtain 108 cells and mixed with four bacteriophage suspensions at a multiplicity of infection of one in separate tubes and incubated for 15 min at 30 °C.

Chorioamnionitis, prolonged ROMs and premature birth have all been associated with MTCT of HIV and may be interlinked [37-39]. However, a Phase III clinical trial of antibiotics to reduce chorioamnionitis-related perinatal HIV-1 transmission showed no benefit in reducing MTCT in the context of single-dose nevirapine prophylaxis [40]. Although both Chlamydia trachomatis and Neisseria gonorrhoeae have been associated with chorioamnionitis, the organisms usually implicated are those associated

with BV, including Ureaplasma urealyticum [41],[42]. A strong association between BV and premature delivery has been reported [43],[44]. There are data from Malawi that suggest that BV may be associated with an increased risk of maternal HIV infection in pregnancy as well as premature delivery and MTCT of HIV [42]. A

study in Gefitinib nmr which mothers received zidovudine from 34 weeks of pregnancy reported that maternal fever >38 °C and BV were associated with in utero transmission of HIV with 2.6-fold and 3-fold risks, respectively [45]. It is not known how applicable this is in settings where mothers receive HAART from earlier in pregnancy. A large meta-analysis assessing the effects of antibiotic treatment of BV in pregnancy does not support the routine screening for, and treatment of, BV in pregnant HIV-negative women [43],[44]. However, the available evidence cannot rule out a small benefit in pregnancy outcome associated with the screening and treatment of BV. The latest Cochrane analysis concludes that there is little evidence that screening and treating Erlotinib price all HIV-1-uninfected pregnant women with asymptomatic BV will prevent preterm delivery (PTD) and its consequences. However, there is some suggestion that treatment before 20 weeks’ gestation may reduce the risk of PTD [46]. In HIV-1-uninfected women, data regarding the effect of screening for and treating BV on premature delivery are conflicting. As outlined above, in HIV-positive pregnant women, there are additional considerations regarding the potential effect of

genital infections on MTCT of HIV-1, but these data are largely from the pre-HAART era. In the setting of full virological suppression on HAART, it is unclear to what extent, if any, Interleukin-2 receptor the presence of any genital infection will contribute to HIV MTCT. Newly diagnosed HIV-positive pregnant women should be screened for sexually transmitted infections as per the routine management of newly diagnosed patients [47]. For pregnant HIV-1-positive women already engaged in HIV care, in the absence of RCTs but for the reasons outlined above, the Writing Group suggests screening for genital tract infections, including evidence of BV. This should be done as early as possible in pregnancy and consideration should be given to repeating this at about 28 weeks. Syphilis serology should be performed on both occasions. In addition, any infection detected should be treated according to the BASHH guidelines (www.bashh.org/guidelines), followed by a test of cure.

cereus group genomes mainly due to the multi-copies of IS231C, IS232A and ISBth166. Taking into account that genome projects usually fail to yield detailed characterization of these elements, we depicted all IS elements in YBT-1520. The disequilibrium in the distribution and copy numbers of ISs among B. cereus group genomes is probably one of the major forces of genome evolution.

Most of these IS elements were probably acquired after the divergence of B. cereus group genomes and contribute to niche adaptation. The study also indicated that the expansion of IS231C in YBT-1520 occurred in evolutionarily recent events due to cycles of expansion and extinction. These data will probably contribute towards further comparative analyses of multiple B. thuringiensis strains, which will shed further light on the impact of ISs transposition on genome diversification. This work was financially supported by

the Chinese National Natural Science Selleckchem Proteasome inhibitor Funds (Grant No. 30930004) and by the National High Technology Research and Development Program of China (863 Program, No. 2008AA02Z112). We are sincerely grateful to Dr Daniel R. Tyrosine Kinase Inhibitor Library Zeigler for the provision of the standard B. thuringiensis strains. We also thank Dr Mick Chandler, Dr Patricia Siguier and Dr Jacques Mahillon for advice on the nomenclature of the ISs we submitted. Table S1. Distribution and number of IS elements on the plasmids of finished Bacillus cereus group genomes. Fig. 1. Phylogeny of IS110 family transposases in Bacillus cereus group genomes. Please note: Wiley-Blackwell is not responsible for

the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Mycoplasma penetrans, a potential human pathogen found mainly in HIV-infected individuals, uses a tip structure for both adherence and gliding motility. Tolmetin To improve our understanding of the molecular mechanism of M. penetrans gliding motility, we used chemical inhibitors of energy sources associated with motility of other organisms to determine which of these is used by M. penetrans and also tested whether gliding speed responded to temperature and pH. Mycoplasma penetrans gliding motility was not eliminated in the presence of a proton motive force inhibitor, a sodium motive force inhibitor, or an agent that depletes cellular ATP. At near-neutral pH, gliding speed increased as temperature increased. The absence of a clear chemical energy source for gliding motility and a positive correlation between speed and temperature suggest that energy derived from heat provides the major source of power for the gliding motor of M. penetrans. Cellular motility is important for a variety of processes, including obtaining nutrients, evading threats, organizing cells for developmental processes, and cell division.

Only 10 patients (20%) had taken any prophylaxis to prevent malaria. Five of these took a drug that was inappropriate for the country to which they traveled. P. falciparum was most common (74%). P. vivax, P. ovale, and P. malariae were present in five, three, and one case, respectively.

In four cases, definitive species identification was not possible due to the low percent parasitemia, with just a few ring forms present. No coinfections were seen. The majority of patients (52%) had parasitemia <1%; only seven patients had hyperparasitemia (>5%). The maximum parasitemia was 28.6%. All cases with >5% parasitemia were P. falciparum. Nonfalciparum forms made up 42% of patients with ≤1% parasitemia EPZ015666 and 12% of those with 1% to 5% parasitemia. Gametocytes were rarely identified. Laboratory results are presented in Table 2. Thrombocytopenia and anemia were the most commonly observed laboratory abnormalities. Mild hyponatremia was also relatively common (36% had sodium ≤135 mEq/L and 12% had sodium ≤130 mEq/L). G6PD levels were measured in 10 children; only one was G6PD deficient. Six patients were tested for sickle cell disease; all were negative. Two patients had known sickle trait. Thirty-four children (68%) were hospitalized for treatment of malaria, with a maximum stay of 9 days. Among those with P. falciparum malaria, 75% were hospitalized;

17% stayed for only 1 day. Documentation of treatment available in 41 children: 18 patients (44%) received quinine and doxycycline, eight (19%)

quinine/quinidine and clindamycin, four (9.7%) received atovaquone–proguanil, six (15%) received only one drug (quinine, choloroquine, Montelukast Sodium or primaquine), and the rest received other Selleckchem SCH727965 combinations. Several children received antibiotic therapy due to concern for additional diagnoses. Sixteen patients had received antimalarial therapy previously, although in some cases this was several months prior. One patient received a blood transfusion for anemia (hemoglobin 5.4 mg/dL). No exchange transfusions were performed. One patient received platelet transfusion for a platelet count of 32,000/ul. All of the patients recovered without serious complications. This case series demonstrates the wide spectrum of possible clinical presentations which may be seen with malaria including vomiting, diarrhea, headache, abdominal pain, etc. Gastrointestinal symptoms can be so severe that an intestinal infection may be suspected. Hepatosplenomegaly may be seen; this was less common in our series than in other reports.14,15 In contrast to the report by Viani and Bromberg,14 hyponatremia was not a common finding. One almost universal symptom is fever, either by history or at presentation. Because malaria may present with a wide variety of clinical symptoms, a high index of suspicion is required to ensure prompt diagnosis. Primary care providers seeing patients with a history of fever should always ask about a history of travel and request the appropriate diagnostic tests.

Complete resolution of the side effect with efficacy has been reported in 72% and 86% of patients treated in this way.[49, 53] Thiopurine-induced pancreatitis occurs in approximately 4% of patients[38] MAPK Inhibitor Library supplier and has been considered a strict contraindication to subsequent treatment with an alternative thiopurine.[75] Three small retrospective case series (< 10 patients each) have examined rechallenging patients with 6MP after AZA-induced pancreatitis, with overall success rates varying from 29% to 100%.[76-78] There are no data regarding the use of allopurinol to overcome thiopurine-induced pancreatitis. Thus, if an adverse event occurs on AZA, it is worthwhile to have a trial of 6MP (initially at low dose) and, if that

fails, then the addition of low-dose allopurinol with 6MP, but only if a recurrence of the adverse event would be tolerated by the patient. If the adverse event occurs on 6MP

as the initial drug, anecdotal experience suggests a trial of AZA may also be worthwhile, followed by combination therapy if unsuccessful. Thiopurines have been the mainstay Proteasome inhibitor of treatment in IBD for many years and have also been extensively used in various rheumatological conditions. With the ability to measure thiopurine metabolites, important strides have been made in the IBD world to improve efficacy and optimize dosing of thiopurines, including in combination with low-dose allopurinol. In IBD, a therapeutic window of 230–260 to 450 pmol/10 × 88 RBCs has been established. Above this level, there are significantly

increased risks of side effects, including myelotoxicity, without any gain in efficacy. BCKDHB Studies in IBD have shown that over 30% of patients who would previously have been declared ‘refractory’ or ‘intolerant’ to thiopurines are now otherwise able to remain on monotherapy with improved clinical outcomes. Much of this work has yet to be undertaken within the rheumatology community. While the upper limit of 6TGN is a relevant threshold to apply in rheumatology due to the risk of universal side effects, the minimum effective 6TGN level is yet to be determined in different rheumatological conditions. The addition of allopurinol should also improve thiopurine metabolic profiles in rheumatology patients who are thiopurine shunters. It may be prudent for a rheumatology patient failing thiopurines to have their metabolites checked prior to drug cessation. “
“Hydrogen sulfide (H2S) is a gaseous mediator produced in the body. In experimental models, endogenously produced H2S has been shown to have pro-inflammatory effects. The aim of this study was to investigate whether H2S is present in three common rheumatic diseases, rheumatoid arthritis (RA), gout and osteoarthritis (OA) and to determine if H2S levels correlate with disease activity. Patients with RA, gout, OA, and healthy controls (n = 30 each) were recruited. Plasma and where possible, synovial fluid (SF), were obtained.

Gibbons et al. (2000) had isolated a gene

from Salmonella responsible for the introduction of a 2-hydroxyl group into a lipid-A-bound myristic acid residue. The hydroxylation reaction is catalyzed by the Fe2+/O2/α-ketoglutarate-dependent LpxO dioxygenase. Rojas-Jiménez et al. (2005) had identified a gene called olsC in R. tropici encoding an LpxO homolog responsible for the synthesis of hydroxylated OLs. Later, it was shown that OlsC is responsible for the introduction of a hydroxyl group in the C-2 position of the piggy-back fatty acid of OLs (Vences-Guzmán et al., 2011). A prediction indicates that OlsC of R. tropici CIAT899 is a water-soluble protein of 281 Selleckchem Inhibitor Library amino acids (Rojas-Jiménez et al., 2005). Owing to its homology to LpxO from Salmonella, it can be expected that OlsC-dependent hydroxylation of the ester-linked fatty acid will also be Fe2+/O2/α-ketoglutarate dependent. Genes encoding OlsC homologs can be found in Selleckchem CX-4945 Agrobacterium vitis, Agrobacterium radiobacter, Ochrobactrum anthropi, Ochrobactrum intermedium, Aurantimonas manganoxydans, Fulvimarina pelagi, Roseomonas cervicalis, Chelativorans sp., Mycobacterium rhodesiae, and several Brucella species (Supporting

Information, Table S1). Interestingly, in the so-called classical Brucella such as Brucella ovis, Brucella suis, Brucella melitensis, or B. abortus, which are intracellular pathogens, the olsC gene is present only as pseudogene containing a frameshift mutation. As a consequence, the olsC gene is translated into two ORFs, making the gene olsC nonfunctional (Palacios-Chaves et al., 2011). In the genomes of several atypical Brucella strains such as Brucella microti, Brucella sp. BO1, or Brucella sp. BO2 which share several characteristics with the opportunistic soil pathogen Ochrobactrum, olsC genes lacking the frameshift can

be detected that are probably functional. This observation implies that organisms like Ochrobactrum, R. tropici, and nonclassical Brucella such as Brucella isolated from soil that present PRKACG both (De et al., 2008;Scholz et al., 2008a, 2008b, 2009, 2010) an intracellular and a free-living lifestyle have preserved a functional copy of olsC, whereas the classical Brucella strains that are strictly intracellular pathogens present only a nonfunctional copy of olsC (Palacios-Chaves et al., 2011). A functional OlsC might confer a selective advantage in adverse abiotic stress conditions, but might not be of use or even have a negative impact when the bacteria are inside a host. Recently, Vences-Guzmán et al. (2011) reported a more detailed study of an olsC-deficient R. tropici mutant. Strains lacking the OL hydroxylase OlsC showed a growth defect at increased temperatures (37 and 42 °C) and under acid pH conditions (4.5 and 4.0).

1b), confirming that the NMA0797/0798 TCS was involved in the induction of the expression of the pilC1 gene during N. meningitidis–host cell interaction (Jamet et al., 2009). Moreover,

in mutant KZ1CNMA1803, the β-galactosidase activity was induced upon contact with host cells at a level significantly higher than that in the wild-type KZ1C strain (P<0.01) (Fig. 1b). To confirm these data, the NMA1803 mutation was introduced into the parental N. meningitidis strain 8013 that is devoid of pilC1-lacZ fusion. Total RNA was isolated from the wild-type 8013 and the 8013NMA1803 mutant strain grown in an infection medium and harvested after 1 and 4 h of adhesion to HUVECs. The level of transcription of pilC1 was measured using real-time quantitative RT-PCR. The results confirmed the

Y27632 increased level of pilC1 induction in mutant 8013NMA1803 compared with that RGFP966 nmr of the wild-type 8013 strain (data not shown). Altogether, these data demonstrated that insertion of a transposon into gene NMA1803 resulted in augmented expression of the pilC1 gene upon contact with host cells. Analysis of the genomic location of the NMA1803 gene revealed that it is located in a putative operon spanning from gene NMA1802 to gene NMA1806 (Fig. 2a). Because gene NMA1802 belongs to the REP2 regulon, which is upregulated upon contact with host cells (Morelle et al., 2003), we first investigated whether the expressions of the genes located downstream 17-DMAG (Alvespimycin) HCl of gene NMA1802, i.e. genes NMA1803, NMA1805 and NMA1806, were also regulated upon interaction with host cells. The level of transcription of genes NMA1802–NMA1806 was determined using quantitative RT-PCR from total RNA isolated from strain 8013 grown in an infection medium and from bacteria adherent

to HEC-1B cells. This revealed that the expression of the four genes was coordinately induced upon contact with host cells (Fig. 2b). Moreover, NMA1803 and NMA1805 are overlapping ORFs (Fig. 2a; Vallenet et al., 2006). Analysis of the cotranscription of adjacent genes by RT-PCR revealed that the adjacent NMA1802–NMA1803 and NMA1805–NMA1806 genes were cotranscribed (Fig. 2c). Altogether, these data demonstrated the operonic organization of genes NMA1802–NMA1806. As a corollary, the REP2 sequence that is located upstream of gene NMA1802 contains a promoter for the whole operon, thus being consistent with the above data showing an upregulation of genes NMA1802–NMA1806 following adhesion onto host cells. According to database annotations, NMA1803 is a pseudogene that is part of a putative two-component system, where NMA1803 is encoding the putative sensor and NMA1805 the putative regulator (Vallenet et al., 2006). The protein encoded by NMA1803 lacks the cytoplasmic transmitter and nucleotide-binding domains found in functional sensors (Snyder et al.

1b), confirming that the NMA0797/0798 TCS was involved in the induction of the expression of the pilC1 gene during N. meningitidis–host cell interaction (Jamet et al., 2009). Moreover,

in mutant KZ1CNMA1803, the β-galactosidase activity was induced upon contact with host cells at a level significantly higher than that in the wild-type KZ1C strain (P<0.01) (Fig. 1b). To confirm these data, the NMA1803 mutation was introduced into the parental N. meningitidis strain 8013 that is devoid of pilC1-lacZ fusion. Total RNA was isolated from the wild-type 8013 and the 8013NMA1803 mutant strain grown in an infection medium and harvested after 1 and 4 h of adhesion to HUVECs. The level of transcription of pilC1 was measured using real-time quantitative RT-PCR. The results confirmed the

Selleck MLN0128 increased level of pilC1 induction in mutant 8013NMA1803 compared with that Napabucasin of the wild-type 8013 strain (data not shown). Altogether, these data demonstrated that insertion of a transposon into gene NMA1803 resulted in augmented expression of the pilC1 gene upon contact with host cells. Analysis of the genomic location of the NMA1803 gene revealed that it is located in a putative operon spanning from gene NMA1802 to gene NMA1806 (Fig. 2a). Because gene NMA1802 belongs to the REP2 regulon, which is upregulated upon contact with host cells (Morelle et al., 2003), we first investigated whether the expressions of the genes located downstream 3-mercaptopyruvate sulfurtransferase of gene NMA1802, i.e. genes NMA1803, NMA1805 and NMA1806, were also regulated upon interaction with host cells. The level of transcription of genes NMA1802–NMA1806 was determined using quantitative RT-PCR from total RNA isolated from strain 8013 grown in an infection medium and from bacteria adherent

to HEC-1B cells. This revealed that the expression of the four genes was coordinately induced upon contact with host cells (Fig. 2b). Moreover, NMA1803 and NMA1805 are overlapping ORFs (Fig. 2a; Vallenet et al., 2006). Analysis of the cotranscription of adjacent genes by RT-PCR revealed that the adjacent NMA1802–NMA1803 and NMA1805–NMA1806 genes were cotranscribed (Fig. 2c). Altogether, these data demonstrated the operonic organization of genes NMA1802–NMA1806. As a corollary, the REP2 sequence that is located upstream of gene NMA1802 contains a promoter for the whole operon, thus being consistent with the above data showing an upregulation of genes NMA1802–NMA1806 following adhesion onto host cells. According to database annotations, NMA1803 is a pseudogene that is part of a putative two-component system, where NMA1803 is encoding the putative sensor and NMA1805 the putative regulator (Vallenet et al., 2006). The protein encoded by NMA1803 lacks the cytoplasmic transmitter and nucleotide-binding domains found in functional sensors (Snyder et al.