In this study, the mutant JX22MT1 was obtained by the EZ-Tn5 tran

In this study, the mutant JX22MT1 was obtained by the EZ-Tn5 transposon mutation and showed no antifungal activity against Fusarium oxysporum f. sp. lycopersici as compared with wild-type strain JX22.

The pqqC gene was disrupted in the mutant. Antifungal activity at the wild-type level was restored from the mutant JX22MT1 with the introduction of the functional pqqC gene, which encodes pyrroloquinoline–quinone synthesis protein C. The results suggest that pqqC is essential for antifungal activity of P. kilonensis JX22 against F. oxysporum f. sp. lycopersici. “
“Consumption see more of Vibrio parahaemolyticus via contaminated shellfish results in inflammatory gastroenteritis characterised by severe diarrhoea, nausea and stomach cramps. This study investigated

the translocation of V. parahaemolyticus across a Peyer’s patch M cell-like Caco-2/Raji B co-culture model system, as M cells represent a primary site of infection for many pathogenic bacteria. Vibrio parahaemolyticus translocated across co-culture monolayers in higher numbers as compared to Caco-2 monolayers. Moreover, the bacteria induced a greater disruption of the transepithelial resistance in M cell-like co-cultures than in Caco-2 monocultures. Virulence factors associated with this pathogen include two type three secretion systems (TTSS-1 and Olaparib cost TTSS-2). TTSS-1 had no effect on translocation efficiency, with TTSS-2 exhibiting a modest enhancing effect. ERK activity was required for optimal translocation 1 h postinfection, however,

neither ERK nor the JNK and p38 MAPK were required at 2 h pi. Additionally, TER disruption in response to bacterial infection occurred independently of the TTSS and MAPK activation. It was concluded that V. parahaemolyticus causes TER disruption of M cell-like co-cultures and translocates in high numbers across the M cell-like co-culture monolayer. These data implicate M cells as important sites for V. parahaemolyticus invasion across the intestinal epithelium during infection. The human gastrointestinal pathogen ID-8 Vibrio parahaemolyticus is a Gram-negative bacterium whose natural habitat is marine and estuarine sediment (Daniels et al., 2000; Makino et al., 2003). Infection is characterised by severe gastroenteritis following consumption of contaminated, uncooked shellfish. Infection of the host epithelium by V. parahaemolyticus is associated with the presence of two haemolysins and two type three secretion systems, namely TTSS-1 and TTSS-2. While TTSS-1 is involved in the cytotoxic effects of the bacterium, TTSS-2 is responsible for bacterial enterotoxicity (Park et al., 2004a, b). The intestinal monolayer is an important defensive barrier following the consumption of contaminated seafood (Catalioto et al., 2011).

In Turkey where diarrheagenic Escherichia coli are the major path

In Turkey where diarrheagenic Escherichia coli are the major pathogens,4,5 the rate of protection against TD with rifaximin observed in this study (67%) was similar to that observed in a prior study by DuPont and colleagues6 among student

travelers to Mexico. The design of this study is unique from previous rifaximin prophylaxis trials because of the higher rifaximin daily dose (1,100 mg) administered. A safe and effective QD dosing regimen of rifaximin would be more convenient and potentially more cost effective versus a twice daily (BID) or three times daily (TID) dosing regimen. Although unclear, one wonders if a higher QD rifaximin dose of 1,100 mg might also have a residual protective impact seen with more frequent daily dosing regimen at lower rifaximin doses (eg, 200 mg BID or TID). Alternatively, QD scheduling at any dose may not be as effective as BID dosing given the possibility of a therapeutic trough with QD dosing, although in the DuPont and Selleckchem AG-14699 colleagues6 study, efficacy was observed with rifaximin 200 mg QD dosing. This study has important limitations including inadequate power due to lower than anticipated attack rate, limited microbiological outcomes, nonsequential treatment allocation, as well as issues of adherence ascertainment and to a lesser extent daily diary completion among enrollees. Despite these deficiencies, there was no discernable effect of the nonsequential treatment allocation on primary outcomes, although

such an effect cannot be ruled out. Furthermore, restricting

analysis to those for whom adequate Ivacaftor adherence and outcome ascertainment could be assessed resulted in no appreciable change in the primary outcome with an estimated protective efficacy 71% (−34% to 94%; Fisher’s exact p = 0.14). Given the potential harms of long-term daily antibiotics in a population at risk for trauma-associated infections (including enteric trauma) and impact on individual and community microbiomes, it is uncertain that antimicrobial chemoprophylaxis would offer a practicable solution during most extended military deployments (which historically have averaged about 3–6 mo). However, Molecular motor there are a number of relevant settings including port visits, in special operations forces, or in the initial phase of deployment settings where risk of TD is highest and the consequences of heat injury are frequent, where chemoprophylaxis may offer an acceptable solution. Further studies to explore the efficacy and safety of TD chemoprophylaxis in these populations and settings are warranted. This study was supported by Salix Pharmaceuticals under a cooperative research and development agreement, and the Department of Defense Military Infectious Disease Research Program (Fort Detrick, MD, USA) under work unit no. 6000.RAD1.D.E0301. One or more authors of this article are military service members (or employees of the US Government). This work was prepared as part of official duties.

The relative bioavailability was assessed by comparing the NVP XR

The relative bioavailability was assessed by comparing the NVP XR and IR trough concentrations at week 24 and the geometric mean of all weeks. In determining the sample size, a planned noninferiority margin of 12% was selected for the difference in proportions between NVP XR and NVP IR in terms of continued virological response, assuming that 90% would be responders in both groups. A noninferiority test, with a one-sided α = 0.025 and a randomization ratio of 2:1 for the NVP XR and NVP IR treatment

arms, required 198 and 99 patients, respectively, in order to Selleckchem Oligomycin A have 90% power to reject the null hypothesis. The primary endpoint (proportion of patients with continued virological response at week 24) and its 95% confidence interval (CI) were estimated based on a time to loss of virological response (TLOVR) algorithm as specified by the US Food and Drug Administration (FDA) guidance [16]. Weighted treatment difference and corresponding variance were calculated Nutlin-3a nmr based on Cochran’s statistic [17] with continuity for variance calculation. Noninferiority to the control group in the primary endpoint was determined by comparing the lower 95% confidence limit of the difference in proportions of virological response for the two treatment arms (NVP XR vs. NVP IR) with the noninferiority margin of −12%. Because of the increased numbers of patients enrolled in this study, the noninferiority

margin for the study was adjusted to −10%. An additional approach (SNAPSHOT analysis) was also used to analyse the endpoint of continued suppression, as a key secondary analysis. In this approach, a patient with VL < 50 copies/mL at the 24-week time-point (± 4) was defined as a virological responder. The secondary endpoint of TLOVR using an LLOQ of <400 copies/mL was analysed using the Cox proportional hazard model with baseline background therapy as a stratum variable. All safety data were analysed using descriptive statistical methods. A total of 499 patients were enrolled in the study, an increase over the planned Etofibrate number of 300. This was a result of the unexpectedly rapid enrolment as a result of investigators pre-screening their patients. Of these, 445 were randomized, 295 to NVP XR and 148

to NVP IR; 54 patients were excluded primarily because they did not meet the eligibility criteria (Fig. 1). Two patients, one in each treatment group, were randomized but never received treatment, leaving 443 in the full analysis set. Baseline demographic data, which are shown in Table 1, were similar for the two treatment groups. The baseline VL value was defined as the mean of the VLs at screening and at randomization; 27 patients had a VL > 50 copies/mL at the randomization visit, so 6.1% of patients had a ‘detectable’ baseline VL. As the results for VL at randomization were not available until several days after randomization, these patients were still included in the study and continued in the study based on the earlier nondetectable screening of VL.

When the monolayer is not disrupted, the recovered CFU mL−1 shoul

When the monolayer is not disrupted, the recovered CFU mL−1 should remain essentially constant over the same

time course. The S. Typhimurium 14028s (black diamonds) and S. Typhimurium 14028s ΔsopD2∷FRT (NT060) (white circles) showed a slight decline over the time course of the assay, suggesting that the monolayer integrity was not significantly affected by these strains (Fig. 3). In contrast, CFU mL−1 of S. Typhi STH2370 abruptly decreased until they became undetectable, strongly suggesting that gentamicin leaked due to a monolayer disruption (white squares). When S. Typhi was complemented with sopD2STM gene (in the pNT007 plasmid, see Materials and methods) and used to infect the monolayer, we observed that the corresponding CFU mL−1 showed a sharp difference with the otherwise isogenic wild-type strain resembling the S. Anti-diabetic Compound Library price Typhimurium phenotype (black triangles). The CFU mL−1 numbers from infected cells with S. Typhi carrying the empty plasmid (pCC1) showed no differences with respect to the wild-type strain (data not shown). It has been reported that sopD2 contributes to the synthesis of Sifs, lipid filaments essential for S. Typhimurium intracellular

proliferation (Brumell et al., 2003; Jiang et al., 2004; Birmingham et al., 2005). When we performed a gentamicin protection assay, we observed that S. Typhi sopD2STM showed a significant decrease of CFU recovered from HEp-2-infected monolayers compared with the wild-type strain (Fig. 4). In contrast, S. Typhi sopD2STM showed similar invasion levels compared with S. Typhimurium 14028s ΔsopD2∷FRT (NT060) (P=0.13749). The results suggest that loss of SopD2 function in the serovar Typhi contributes to the bacterial intracellular proliferation in human epithelial cells. In the process of adaptation to humans, bacterial genes no longer compatible with the lifestyle of facultative

pathogens within the host are selectively inactivated. These inactivated genes are called ‘antivirulence genes’ and their loss of function results in the adaptation to a given host (Maurelli, 2007). Salmonella enterica serovar Typhi is a facultative bacterial pathogen that has accumulated a large number of pseudogenes (approximately 5% of the genome), over 75% of which have completely lost their function (McClelland et al., 2004; Urease Dagan et al., 2006). Compared with free-living organism genomes, facultative pathogens harbor several pseudogenes and a gene population structure that promotes the maintenance of specific mutations. In contrast to free-living bacteria (large genomes, a great diversity of functional genes and low percentage of laterally transferred genes) and obligate parasites (extremely reduced genomes), S. Typhi represents an intermediate step exhibiting some genome erosion directed to inactivation and loss of detrimental or nonessential functions for its environment, i.e. the host (Ochman & Moran, 2001).

The objectives of the study were to investigate whether the risk

The objectives of the study were to investigate whether the risk of developing lymphoma was Vemurafenib purchase increased when blood EBV DNA load was high in preceding years and whether a cut-off value above which patients would be at a very high risk of progression to ARL could be determined. We conducted a nested case–control study within the French ANRS PRIMO and SEROCO/HEMOCO cohorts. Cases of B lymphoma were classified into two different groups: systemic B lymphoma and PBL. Ethics committee approvals were obtained for the two cohorts (PRIMO and SEROCO/HEMOCO)

and all patients gave written consent to participate in the cohort. Between 1996 and 2009, 808 antiretroviral-naïve HIV-infected patients presenting at the time of primary infection were enrolled in the ongoing ANRS PRIMO cohort. Primary infection was confirmed by an incomplete Western blot, or a positive p24 antigenaemia or a detectable plasma viral load with a negative or weakly reactive enzyme-linked immunosorbent assay (ELISA) test, or an interval of less

than 6 months between a negative and a positive ELISA test [17]. In this cohort, sera were collected and frozen at −80°C every 6 months and cells were collected and frozen at −196°C every 12 months. In the ANRS SEROCO/HEMOCO cohort, 1748 HIV-infected patients who had a recent diagnosis of HIV-1 infection (< 1 year) or a well-documented date of seroconversion were enrolled between 1988 and 2001 [18]. Serum samples and PBMC samples were collected and stored at −196°C every 6 months and every 18 months, respectively. In these cohorts, visits were Pifithrin-�� in vitro almost scheduled for clinical and biological examination every 6 months. The occurrence of AIDS-related events was recorded at follow-up visits (reviewed and checked in the medical files) and through repeated cross-checking with the national AIDS registry. At the time of this analysis, a diagnosis of NHL/brain lymphoma had been reported in 72 patients. Among these patients

with lymphoma, 43 patients, including 29 with B NHL confirmed histologically and 14 with PBL for whom no histology was available, had available frozen PBMCs and/or serum samples collected within 3 years preceding the diagnosis of lymphoma. EBV-encoded small RNA (EBER) mRNA results were not available except in one case of systemic B NHL, in which EBER mRNA was detected. The date of diagnosis of the lymphoma was called the ‘index date’. For each case, two controls were randomly selected from eligible individuals included in the cohorts with the same CD4 count (± 30 cells/μL) in the year of lymphoma diagnosis (index case). The main known risk factors for NHL (CD4 cell count and age) were taken into account by adjustment in multivariate analysis. Characteristics of the cases and controls are reported in Table 1.

, 2006; Jones & Dangl, 2006) PTI is induced by perception of pat

, 2006; Jones & Dangl, 2006). PTI is induced by perception of pathogenic PAMPs with specific plant cell surface pattern-recognition receptors (PRRs). Flagellin Sensing 2 (FLS2) is one of the best characterized PRRs, and specifically perceives a highly conserved 22-amino-acid peptide flg22 derived from the amino terminus of

Pseudomonas syringae flagellin (Felix et al., 1999; Chinchilla et al., 2006). Perception of flg22 usually triggers mitogen-activated protein kinase (MAPK) activation, transcription of resistance-related genes, reactive oxygen species (ROS) production, and callose deposition (Felix et al., 1999; Asai et al., 2002; Nicaise et al., 2009). ETI is activated by plant intracellular resistance (R) proteins after specific perception of pathogenic T3SEs. It is often associated with a hypersensitive response (HR), a form Akt inhibitor of rapid programmed cell death at the site of infection learn more (Takken & Tameling, 2009). In most cases, R proteins recognize effectors through monitoring specific host proteins, which are

targeted and modified by pathogen effectors. For example, P. syringae secreted effectors AvrRpt2 and AvrRpm1 target and cause Arabidopsis RPM1-interacting protein 4 (AtRIN4) cleavage and phosphorylation, respectively, and the modifications of AtRIN4 are then monitored by R proteins RPS2 and RPM1, resulting in a rapid initiation of ETI (Mackey et al., 2002, 2003; Axtell & Staskawicz, 2003). Dozens of T3SEs have been identified from Pseudomonas species, and most of them can suppress plant PTI and/or ETI responses (Guo et al., 2009). One of these effectors, HopF2,

was recently reported to target and ADP-ribosylate both MAPK kinase 5 (MKK5) and RPM1-interacting protein 4 (RIN4) in Arabidopsis to block PTI and AvrRpt2-trigerred ETI (Wang et al., 2010; Wilton et al., 2010). HopF1 (also named AvrPphF) is a homolog of HopF2 in P. syringae pv. phaseolicola (Psp), a bean pathogen (Tsiamis et al., 2000). HopF1 triggers cultivar-specific resistance in bean plants (Phaseolus vulgaris) containing the R1 disease resistance gene and promotes virulence in plants lacking the resistance gene (Tsiamis et al., 2000). Although HopF1 was cloned more than a decade ago, the real virulence and avirulence targets of this effector remain unclear. HopF1 shares about 48% amino acid sequence identity with HopF2, which was confirmed Levetiracetam to be an active ADP-ribosyltransferase (ADP-RT). Although no ADP-RT activity was detected in a standard in-vitro assay, HopF1 owns the same putative ADP-RT active sites with HopF2, and these sites are necessary for the virulence and avirulence functions of this effector in bean (Singer et al., 2004). Thus, RIN4 and MKK5 homologs of bean are possibly the virulence or avirulence target of HopF1. Due to the technical challenge of transformation, a long growth cycle and lack of complete genomic information, studies about the gene functions of bean are not well developed.

Purified VDHs showed activities toward some aromatic

Purified VDHs showed activities toward some aromatic selleck chemicals llc aldehydes. These enzymes have the same subunit molecular mass of about 57 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but differed in some of their observed properties. Native molecular masses also differed between the purified enzymes. These were 250 kDa for the enzyme from alkaliphilic strain TA1 and 110 kDa for that from neutrophilic strain TM1, as determined by gel filtration. The enzyme from strain TA1 required NADP+ as a coenzyme for its activity, but that from strain TM1 required NAD+. These results are important because this is the first report of an alkaliphilic bacterium consuming lignin monomers. Vanillin

(4-hydroxy-3-methoxybenzaldehyde) is one of the most important aromatic flavor compounds widely used in the food industry and in fragrances for perfumes. This compound is extracted from the fermented pods of Vanilla orchids. However, only about 0.2% of the market demand for ICG-001 vanillin is met by extraction from Vanilla pods (Krings & Berger, 1998). This natural vanillin is more expensive than the synthesized compound (Priefert et al., 2001). There is an increasing interest

and demand for natural vanillin from consumers. In the United States and European Union, the term ‘natural’ can be applied to a product that is derived from a natural raw material via biological conversions using enzymes or whole cells (Venkitasubramanian et al., 2008). Because of this, numerous studies on natural vanillin biosynthesis using microorganisms or enzymes have been conducted. Several potential feedstocks, including curcumin, Siam benzoin resin, phenolic stilbenes, eugenol, and ferulic acid, have been suggested for the production of vanillin (Ghosh Glycogen branching enzyme et al.,

2007; Unno et al., 2007; Yamada et al., 2007). However, these bioconversions are not yet economically feasible. A genetic approach to metabolic engineering has also been developed for the production of vanillin from eugenol and ferulic acid. Because some Pseudomonas strains metabolize these compounds with vanillin as an intermediate, the inactivation of vanillin dehydrogenase (VDH) enzyme by making a null mutant of the gene for vanillin accumulation has been investigated (Overhage et al., 1999). Vanillin (molar yield of 44.6% relative to the initial eugenol concentration) is obtained from eugenol by blocking vanillin catabolism in the mutant. These metabolic engineering approaches can be effective for vanillin production. Therefore, we attempted a novel approach for producing vanillin using microorganisms or their genetic mutants. Because a high concentration of ferulic acid can be dissolved under alkaline conditions (≥150 g L−1 at pH 10 vs. ≤15 g L−1 at pH 7 in our simple solubility examination), we screened an alkaliphile that can grow on ferulic acid as the sole carbon source.

Then, cells were incubated with FITC-conjugated anti-rabbit IgG 1

Then, cells were incubated with FITC-conjugated anti-rabbit IgG 1 : 50 for 1 h at 37 °C. Fluorescence at 525 nm was measured in a microplate reader Spectramax M2e (Molecular Devices, Sunnyvale). Conidia macerated with liquid nitrogen were used as a sample of the total (extra- and intracellular) GAPDH protein. Conidia without

an immunolabeling treatment were used as the negative control. Conidial suspensions of a wild-type (WT) green fluorescent protein expressing M. anisopliae (2 × 107 conidia mL−1) were used to treat insect wings by immersion for 20 s. The wings see more from Dysdercus peruvianus disinfected previously in 37% H2O2 were placed on the surface of 0.7% water agar and incubated for 8 h at 28 °C to induce conidial swelling and germination. The conidia were counted in five objective fields under a fluorescence microscope and recorded with three replicates of wings. The conidia were counted before VE821 and after washing in 0.05% Tween 20 for 30 s. The following treatments were performed: (1) before immersion of the wings in the conidial suspension – preincubation with bovine serum albumin (BSA) (25 μg mL−1) for 1 h at 37 °C and preincubation with recombinant

GAPDH (25 μg mL−1; from M. anisopliae, Supporting Information, Appendix S1) for 1 h at 37 °C; (2) conidial suspension was treated before immersion of the wings – with anti-CHI2 antisera (1 : 100) for 1 h at 37 °C and anti-GAPDH antiserum (1 : 100, produced with P. brasiliensis GAPDH). Experiments were in triplicate; the means and SEs were determined. Statistical analysis was performed using a t-test. P values of 0.0001 or less were considered statistically significant. The ORF from gpdh1 (GenBank accession number EF050456) predicts a 338 amino acid protein with an estimated MW of 36 kDa and ID-8 a theoretical pI of 8.26. In silico protein domain analysis found no domain other than the expected NAD-binding domain (from Val4 to Cys151) and the C-terminal domain (from Leu156 to Tyr313) typical of GAPDH (Figs 1 and S1; Appendix S2). The putative M. anisopliae GAPDH sequence

had high identity and similarity values with fungal counterparts (Table S1), and a phylogenetic tree was built (Fig. S2) showing a distribution consistent with other orthologs and one intron at a conserved position (Ridder & Osiewacz, 1992; Templeton et al., 1992; Jungehulsing et al., 1994). The M. anisopliae gpdh1 gene is a single copy (Fig. 1a). To characterize possible isoforms of the GAPDH in M. anisopliae, cell extracts were analyzed by 2-D gel electrophoresis [Fig. 1b (A)]. The immunodetection of native GAPDH isoforms was performed using anti-GAPDH P. brasiliensis polyclonal antiserum. The Western blot revealed three reactive isoforms, with pIs of 6.6, 6.8 and 7.0 [Fig. 1b (B)]. A protein with a molecular mass of 36 kDa and pI 7.0 was excised from 2-D gel electrophoresis blots of M. anisopliae mycelial protein extracts.

The canyon – a rectangular cross-section tube – lay in the surfac

The canyon – a rectangular cross-section tube – lay in the surface of a schematic planet. In the canyon, there were three types of spaceship marked by different colors (blue, red, and green). The color of the controlled spaceship was blue. That was directed with the gamepad along the horizontal dimension of the canyon. In every second, one spaceship appeared at the start of the canyon and moved towards the blue spaceship. The color of the spaceship was red with 0.6 probability and green with 0.4 probability.

The aim of the task was to avoid the red spaceships and to catch the green ones with the controlled spaceship. To perform the task properly, participants Sunitinib had to fixate in the location where the spaceships appeared. For more details, see Sulykos & Czigler (2011). Electroencephalographic

activity was recorded (DC, 70 Hz; sampling rate, 500 Hz; Synamps2 amplifier, NeuroScan recording system) with Ag/AgCl electrodes placed at 61 locations according to the extended 10–20 system by use of an elastic electrode cap (EasyCap). The reference electrode was on the nose tip, and offline re-referenced to the average activity.‎ Horizontal electrooculographic buy Y-27632 activity was recorded with a bipolar configuration between electrodes positioned lateral to the outer canthi of the eyes. Vertical eye movement was monitored with a bipolar montage between electrodes placed above and below the right eye. The electroencephalographic signal was bandpass-filtered offline, with cutoff frequencies of 0.1 and 30 Hz (24-dB slope). Epochs of duration 600 ms, including a 100-ms prestimulus interval, were extracted for each event, and averaged separately for the standard and deviant stimuli. The mean voltage during the 100-ms prestimulus interval was used as the baseline for amplitude measurements, and epochs with an amplitude change exceeding ± 50 μV on any channel were excluded from further analysis. Event-related potentials were averaged separately

for the standard and deviant stimuli (symmetric and random) in the two conditions. Responses to the third to the seventh standards after a deviant were included in the standard-related ERPs. To identify change-related activities, ERPs elicited by standard stimuli were subtracted filipin from ERPs elicited by deviant stimuli in the reverse condition. Note that, in many studies, vMMN was calculated as the difference between the ERPs elicited by deviant and standard of the same stimulus sequence. With this method, the effect of physical differences between the deviant and standard and the effect of memory-related mismatch effects are confounded. Therefore, comparison of ERPs elicited by identical stimuli is highly recommended (Kujala et al., 2007). Furthermore, comparison of physically identical stimuli (presented frequently/infrequently) in different conditions will not be sufficient to get rid of refractoriness effects adding to plain memory-related effects (Kimura et al., 2009).

The TREAT Asia (Therapeutics Research, Education, and AIDS Traini

The TREAT Asia (Therapeutics Research, Education, and AIDS Training in Asia) HIV Observational Database (TAHOD) is a multicentre prospective cohort of HIV-infected patients, established since September 2003. Data are shared with the International Epidemiologic Selleck Alectinib Databases to Evaluate AIDS (IeDEA). One objective of TAHOD is to evaluate the natural history of HIV disease in ARV-experienced and -naïve patients in the Asia-Pacific region. Seventeen clinical sites (see Appendix A) are included in TAHOD based upon capacity to fulfil data submission requirements and with a view to retaining sites representative

of the region [5]. Ethics approvals were obtained from local Institutional Review Boards and each site sequentially enrolled approximately 200 patients.

Where available, sites provided retrospective data for enrollees and clinical interventions and testing procedures were implemented according to selleckchem local practices. Average follow-up for TAHOD patients in the 12-month period from September 2005 to September 2006 was 86%. Since not all TAHOD patients are taking ARVs, our sampling frame was HIV-infected patients initiating HAART, any combination of three or more ARVs, from 2000 onwards. Eligible patients were also required to have at least one subsequent clinical visit or result recorded in the database, post-therapy, at the time of analysis. Patient covariates included demographics (age at entry to cohort, gender, HIV source exposure), indices of illness severity [Centers for Disease Control and Prevention (CDC) classification, baseline CD4 lymphocyte count and HIV RNA], hepatitis B and C coinfections and prescribed HAART regimen. Retrospective and prospective data were included. The CDC classification for TAHOD was modified from the 1993 Center for Disease Control and SPTLC1 Prevention case definition in that it does not differentiate between presumptive and definitive diagnoses

[17]. The most severe pre-HAART CDC category recorded was used as the baseline clinical status. Hepatitis B (C) positive status was defined as being HBsAg (HCV-Ab) positive and patients were assumed to be coinfected for the duration of follow-up. HIV RNA copies/mL and CD4 cell counts up to 91 days prior to HAART initiation were considered for inclusion as baseline values. Where multiple assay results existed, the value closest to the target date was selected. For classifying TAHOD sites with respect to clinical site resourcing, the four-category World Bank criterion (gross national income per capita) was dichotomized into high (upper-middle and upper: >USD 3705) and low (lower-middle and lower: ≤USD 3705) [18]. The annual frequencies of VL and CD4 monitoring of patients reported between December 2006 and February 2007 were also included as measures of site resourcing.