4). Primer extension of mRNA isolated from strain 8013 grown in broth and harvested after adhesion to HUVECs revealed one major mRNA end-point (Fig. 5). Transcription was initiated at the residue G located 55 nucleotides upstream of the translation initiation codon of NMA1803 and separated from the putative −10 box (TATTA) by nine nucleotides (Fig.

5). This finding confirms that NMA1805 displays two promoters: one located in the REP2 sequence and one present in the 5′ end of NMA1803. We also investigated whether NMA1805 bound to one of the four pilC1 promoters (Fig. 1a). None of them were shown to interact with protein NMA1805. BIRB 796 research buy In this work, we explored the regulation of the N. meningitidis pilC1 gene. We identified the protein NMA1805 as a novel regulator involved in

the transcriptional control of pilC1. Perception and response to environmental stimuli are frequently mediated by TCSs (García Véscovi et al., 1996; Beier & Gross, 2006). Classical TCSs consist of a membrane-bound sensor kinase and a cytoplasmic response regulator. The sensor is autophosphorylated in response to an environmental signal. Then, the transfer of the phosphoryl group to the response regulator results in modification of gene expression. Indeed, NMA1805 is annotated as a putative regulatory protein of the NMA1803/1805 putative two-component system (Vallenet et al., 2006). However, NMA1803 has recently been annotated as a nonfunctional truncated sensor (Snyder et al., 2005). Therefore, NMA1805 Morin Hydrate cannot function as a part of this TCS, but can check details still act as a transcription factor because it retains a helix-turn-helix motif allowing DNA binding. Moreover, in this work, we demonstrate a role for NMA1805 in pilC1 regulation. Many other orphan regulators, i.e. without a cognate sensor, have been described previously such as PmrA in Francisella

novicida (Mohapatra et al., 2007) and DegU in Listeria monocytogenes (Gueriri et al., 2008); both are required for bacterial virulence. The absence of a cognate sensor raises the question of signal perception. NMA1805 belongs to the REP2 regulon, and as a corollary, is regulated by the two-component system MisR/S (Jamet et al., 2009). We hypothesized that the operonic organization, under the control of the REP2 promoter, has eliminated the need for NMA1805 to be activated through the perception of a signal by a cognate sensor. Both pilC1 and NMA1805 belong to the REP2 regulon. During the early interaction with host cells, both genes are induced and then NMA1805 is able to induce its own transcription with binding to its own promoter. This work, together with our previous findings, demonstrates that NMA1805 and MisR are necessary to induce pilC1 upregulation upon contact with host cells. Because NMA1805 does not bind to the pilC1 promoters, the precise regulation pathway and the potential collaboration of proteins MisR and NMA1805 are still to be elucidated.

On the basis of these results and comparative genomic studies, we classified the Bf7 phage to the subfamily of Autographivirinae, φKMV-like phages. Pseudomonas

tolaasii is a Gram-negative mushroom pathogenic bacterium that is well known as the causative agent of the yellowing of oyster mushroom (Pleurotus ostreatus) and the Proteasome inhibitor brown blotch disease of champignon, Agaricus bisporus (Bessette et al., 1985; Lee & Cha, 1998). The mushroom infecting phenomenon was firstly described by Tolaas (1915). The bacterium produces a cellular membrane destructive toxin called tolaasin, which disrupts the membrane of the mushroom via pore formation (Rainey et al., 1992). Moreover, bacterial blotch diseases can be caused by other fluorescent pseudomonads such as Pseudomonas agarici, Pseudomonas buy LGK-974 costantinii, Pseudomonas gingeri (Geels et al., 1994; Munsch et al., 2002), and some Pseudomonas fluorescens bv. V strains (Sajben et al., 2011). The infection processes have different characteristics, but the final

result is usually the same: the product becomes unsuitable for sale resulting in serious economic losses. Different studies investigated the significance of fluorescent pseudomonads in the detrimental processes during cultivation and the discolorations after harvesting in case of A. bisporus, Pleurotus pulmonarius, and Lentinula edodes (Thorn & Tsuneda, 1996; Wells et al., 1996). There is an increasing need for appropriate control as the application of most chemical substances during cultivation is prohibited. There are numerous promising investigations for the inactivation of the browning processes with antagonistic bacteria (Fermor & Lynch, 1988; find more Tsukamoto et al., 1998) and toxin neutralizing substances (Soler-Rivas et al., 1999; Tsukamoto et al., 2002). At the same time, there are some Pseudomonas species that play an essential role in sporocarp formation and healthy development of mushrooms (Rainey, 1991), so the complete exclusion

of the genus from cultivation is not a possible option. According to this, the targeted application of bacteriophages as biocontrol agents against these pathogens has great potentials. Phages are viruses of bacteria, and they are ubiquitous in the environment. They play a key role in controlling the bacterial number in different habitats and participate in gene transfer between bacteria (Ashelford et al., 1999). They have great potential as biocontrol agents because of their ability of replication and amplification. Phages cannot be degraded by enzymes; furthermore, they are highly specialized to their host (Goodridge & Abedon, 2003). However, it should be noted that problems may emerge from the rapid development of phage resistant bacterial strains (Guillaumes et al., 1985; Munsch & Olivier, 1995). Several studies have been carried out to isolate bacteriophages against different fluorescent pseudomonads causing diseases (e.g.

Several studies have evaluated protease inhibitor (PI) monotherapies as a maintenance strategy for patients with suppressed HIV viraemia, and shown that viral suppression Selleck Galunisertib can be maintained in over 80% of cases without viral resistance emergence

in the event of viral rebound, with a potential benefit in terms of peripheral fat tissue evolution [11-14]. Two recent randomized studies have investigated darunavir/ritonavir (darunavir/r) as a maintenance strategy. The MONET study demonstrated, at week 48, the similarity of darunavir/r monotherapy to standard triple therapy consisting of darunavir/r plus two NRTIs, with darunavir/r monotherapy having an efficacy rate > 85% [15]. Similarly, the MONOI-ANRS 136 study has shown a high efficacy rate, with HIV viral loads maintained below 400 HIV-1 RNA copies/ml in 99% of the per protocol population receiving a darunavir/r triple-drug regimen compared with 94% of those receiving darunavir/r monotherapy [16]. Because the majority Nivolumab of patients included in the MONOI study received treatment with a nonthymidine nucleoside analogue backbone, and because darunavir/r has been associated with

a good tolerability profile [17-22] in both naïve and experienced patients, it was important to evaluate whether darunavir/r monotherapy could be beneficial in terms of fat distribution and metabolic parameters in long-term HIV-infected patients. Therefore, the aim of the MONOI-ANRS 136 body composition substudy was to evaluate the evolution of body fat composition over 96 weeks in the two treatment strategy groups, namely darunavir/r monotherapy and darunavir/r triple therapy with two NRTIs. The MONOI-ANRS 136 study enrolled adult HIV-infected patients who had been on a stable triple-antiretroviral drug regimen for at least 18 months and who had suppressed viraemia, defined as HIV-1 RNA <400 copies/mL for the previous 18 months, and <50 copies/mL at screening. Patients also had a CD4 count nadir >100 cells/μL and no virological failure during treatment with a prior

PI-containing regimen, and no prior HIV-related neurological disease. Phenylethanolamine N-methyltransferase Patients were recruited from 32 clinical sites in France. The protocol was approved by the Ethics Committee of the Pitié-Salpêtrière Hospital and the French Health Product-Safety Agency (AFSSAPS). All patients provided written informed consent. MONOI was a multicentre, randomized, comparative, 96-week open-label trial that had a primary endpoint of efficacy at week 48. After an initial phase of 8 weeks, during which each patient received darunavir/r at 600/100 mg twice daily in combination with two NRTIs, patients were randomly assigned, 1:1, to either continue the triple-drug darunavir-containing regimen (darunavir/r triple therapy) or discontinue the two NRTIs (darunavir/r monotherapy).

Conflicts of interest: SV has received travel grants from Abbott, BMS, Boehringer-Ingelheim, Gilead Sciences, Roche, Tibotec and ViiV Healthcare.

FW has received travel grants from Abbott, BMS, Boehringer-Ingelheim, Gilead Sciences, Roche, Tibotec and ViiV Healthcare. GM has received research grants from Abbott, Ardea Biosciences, Bionor, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Merck, Pfizer, Theratechnologies and Tibotec. He has received honoraria as a speaker and/or advisor from Astellas, Boehringer-Ingelheim, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Selleck Ruxolitinib Merck, Pfizer, Theratechnologies, Tibotec and ViiV Healthcare. FR received research funding or honoraria from, or consulted for, Bristol-Myers Squibb, Gilead Sciences and Roche. DJ has received

research grants from Tibotec, ViiV and Vertex. He has received honoraria as a speaker from Tibotec, BMS, Merck, BIPI and Gilead and as a consultant for Tibotec, Roche and Gilead. SM has received research grants from Roche and has served on advisory boards for ViiV, BMS and Gilead. He has received honoraria as a speaker from Roche, Tibotec, BMS and Gilead. CK has received travel grants, conference fees or consultancy fees from Abbott, Bristol-Myers Squibb, Gilead, GlaxoSmithKline, Janssen and MSD. MF has served as a scientific advisor for, and/or received honoraria for speaking engagements from, Abbott, Boehringer selleck kinase inhibitor Ingelheim, Bristol-Myers Squibb, Gilead, GlaxoSmithKline, Theratechnologies, Methisazone Tibotec and ViiV Healthcare. LS has received lecture fees, travelling expenses and payment of registration fees from Roche, Bristol-Myers Squibb, MSD, Gilead and Boehringer Ingelheim. DH has received research grants from Pfizer, Tibotec, Gilead and Bionor, and honoraria for advisory boards/consulting from Tibotec, Pfizer, ViiV, Gilead, Monogram and Merck. EDJ has served on the speakers bureau for Gilead Sciences, Merck, Tibotec and Virco and has received honoraria for consulting from Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Merck, Tibotec, and Vertex Pharmaceuticals. He has received research support from

Abbott Laboratories, Achillion, Avexa, Boehringer Ingelheim, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Hoffman LaRoche Laboratories, Merck, Pfizer, Schering Plough, Taimed, Tobira, Tibotec and Vertex. PR has served as a scientific advisor to Boehringer Ingelheim Pharmaceuticals, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Merck & Co., Theratechnologies, Tibotec Therapeutics and Tobira Therapeutics. He has served on data and safety monitoring boards and endpoint adjudication committees for Tibotec Therapeutics and has received honoraria for speaking engagements at scientific conferences from Boehringer Ingelheim Pharmaceuticals, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline and Theratechnologies.

The median time to suppression was 4.55 months (IQR 2.99–7.89 months). In multivariate analyses, older age, male sex, treatment in Ontario rather than British Columbia, non-IDU history, and having an AIDS diagnosis at baseline predicted increased likelihood of suppression. Patients with low baseline viral load were more likely to have suppression [4–5 log10 copies/mL, hazard ratio (HR) 1.27, 95% confidence interval (CI) 1.18–1.38; <4 log10 copies/mL, HR 1.49, 95% CI 1.32–1.68] than patients with baseline viral load ≥5 log10 copies/mL; however, this effect ceased after 18 months of follow-up. Suppression was more likely with nonnucleoside reverse transcriptase inhibitors and ritonavir-boosted

HAART. Identification of patients at risk for diminished likelihood of virological suppression will allow focusing of efforts and selleck kinase inhibitor the utilization of resources to maximize the benefits of HAART. It is well documented that highly active antiretroviral therapy (HAART) decreases morbidity and mortality amongst HIV-positive individuals [1–4]. In particular, one of the primary goals of HAART is the obtainment and maintenance of complete HIV RNA suppression [5]. Failure to achieve and maintain suppression can result in the development of drug resistance and also increases the risk of both horizontal and vertical viral transmission [6–8]. When first initiating

antiretroviral therapy, the obtainment of viral load suppression is an important objective that is associated with a variety of socio-demographic and baseline clinical factors [9,10]. Additionally, choice of initial antiretroviral regimen plays selleckchem an important role in the time it takes to achieve a viral load that Sirolimus research buy is undetectable [11,12]. While the long-term clinical benefits of earlier suppression are unclear, achievement of suppression earlier reduces the length of time one

carries detectable virus and may allow more immediate CD4 T-cell reconstitution [13]. Identifying factors that predict time to virological suppression on modern HAART regimens is thus vital to optimizing therapeutic success. Moreover, identifying such factors will provide useful information to improve care and inform therapeutic treatment guidelines for socio-demographic or clinical risk groups across Canada. This study constitutes the first examination of such factors in the Canadian context with patients initiating modern HAART regimens (defined as first initiation after 2000 with a combination of at least three individual antiretroviral agents). Here we investigate the time to virological suppression and factors associated with suppression among a national cohort of individuals on HAART in Canada. The Canadian Observational Cohort (CANOC) Collaboration is a Canadian cohort study of HIV-positive patients with no prior antiretroviral experience initiating HAART on, or after, 1 January 2000.

, 2000) for auxotrophy. No auxotrophs were detected in our screen of >14 000 clones (∼11 000 of which were recovered following ciprofloxacin treatment as described in Materials and methods). Transposition of phage Mu from its original location to other regions of the chromosome occurs following induction, and is followed by lysis. If the same is true for ECA41, this would explain why the auxotrophy screen failed to yield any colonies: ECA41 may transpose into genes essential for growth on minimal medium, but such cells may not be detected as lysis would follow shortly after. Inward-reading primers flanking the prophages were

designed to detect prophage-less genomes. No ECA41-deficient genomic template was detected, and this is consistent with the stable lysogeny and replicative transposition Daporinad research buy characteristics of Mu. In contrast, an amplicon corresponding to loss of ECA29 was obtained (data not shown). This

amplicon was sequenced, confirming the absence of ECA29 as well as the sequence of the 13-bp direct repeats (GTCAGTAATCGGT) that contribute to the att sites. Navitoclax In contrast with ECA41, excision was precise, reforming the disrupted pflA gene. Spontaneous induction of prophages can result in the presence of phage particles in the culture supernatants of bacterial lysogens. In an attempt to detect these, a filter-sterilized supernatant of a Pa overnight culture was spotted on top agar lawns of 32 different Pa strains, as well as representative strains of E. coli, C. rodentium, Y. enterocolitica and Serratia. No lysis was seen in any case (data not shown). Those Pa strains that carry the prophages are expected to be immune to superinfection by the same phage. Nonetheless, Cobimetinib most of the strains appeared to be naïve to the respective phages (Fig. 1), and could therefore be, in principle, sensitive to infection by these

phages, assuming that the receptor was present. This suggests that no virions were present. Indeed, phage particles were not observed by transmission electron microscopy of culture supernatants following ciprofloxacin treatment (data not shown). It is possible that even though the prophages can excise from the genome, the DNA cannot be packaged into capsids to produce functional virions. Attempts to induce prophage excision and cell lysis using the DNA-damaging agents mitomycin C and UV irradiation were unsuccessful (data not shown). Taken together, these data show that excision of both prophages from the genome occurs, but is a rare, spontaneous and noninducible event, consistent with the relatedness of ECA29 and ECA41 to phage P2 and Mu, respectively. The lysogenic state is stable in these cases, spontaneous induction is rare and they are not inducible by chemical or physical means (Nilsson & Haggard-Ljungquist, 2006; Paolozzi & Ghelardini, 2006).

Both afferents converge onto dendritic spines, the critical site for synaptic integration in MSNs. In advanced PD there is a marked atrophy of dendrites and spines in these neurons, Androgen Receptor activity inhibition indicative of dysfunctional signal integration in the striatofugal pathway. Similar pathology, triggered by a dysregulation of intraspine Cav1.3 L-type Ca2+ channels (Day et al., 2006), has been observed in rodent and primate models of

PD (Day et al., 2006; Neely et al., 2007; Scholz et al., 2008). The significance of such dendritic atrophy and spine pruning for the symptoms and the treatment of PD has remained poorly understood. However, there is increasing awareness that these morphological alterations represent a major obstacle for therapeutic approaches

to enhance striatal function (Schuster et al., 2009). Most notably, the efficacy of dopamine cell replacement strategies, designed to restore nigrostriatal connectivity, may be hampered by striatal dendritic and spine PLX4032 mouse atrophy. In order for grafted dopamine neurons to re-establish functional connections, the morphological target of such reinnervation would need to be preserved or reestablished. In this issue of EJN, Soderstrom et al. (2010) report the results of a study on the impact of dendritic spine preservation in MSNs upon both anti-parkinsonian and prodyskinetic effect of fetal mesencephalic cell grafts. The authors elegantly and convincingly Methocarbamol show that administration of the L-type Ca2+ channel blocker nimodipine prevented loss of spines in MSNs in unilaterally lesioned rats that were grafted with embryonic ventral midbrain cells. Nimodipine treatment also resulted in improved therapeutic benefit and reduced graft-induced behavioral abnormalities of these hemi-parkinsonian rats. Specifically, the results indicate

that graft-mediated anti-parkinsonian efficacy was not potentiated by the prevention of spine loss; however, the impact of the graft- and levodopa-induced side-effects was greatly diminished by nimodipine treatment. Interestingly, these effects were not due to increased survival of grafted cells but correlated with a greater reinnervation of the affected striatum. These results underscore the importance of prevention (or reversal) of spine loss in striatofugal neurons for effective therapy based on dopamine cell replacement. They extend a previous report of reduced levodopa-induced dyskinesia by prior treatment with L-type Ca2+ channel antagonists (Schuster et al., 2009). The results described in Soderstrom et al. (2010) suggest that unless MSN spine loss and dendritic atrophy are reversed by appropriate pharmacological treatment, therapeutic interventions may be of limited efficacy or even cause unwarranted outcome. The findings and conclusions from the study by Soderstrom et al.

1a) and Southern blotting (not shown). Sequence analysis of five of these argR− mutants showed a five amino acid insertion (GVPLL) between the 149th selleck compound and the 150th residue of ArgR (Fig. 4). These mutations all mapped to the terminal α-6 helix of the protein, which we named ArgR5aa. An ArgR derivative

truncated at position 150 was constructed by site-directed mutagenesis. This truncated protein, called ArgR149, was tested for the ability to resolve pCS210 in the argR− strain (DS956/pCS210). ArgR149 displayed the same properties as ArgR5aa, the protein containing the GVPLL insertion between the 149th and the 150th residue, namely a significant reduction in cer site-specific recombination in vivo (Fig. 1b) and the ability to repress an argA∷lacZ fusion in vivo. In order to quantify the levels of repression of the argA∷lacZ fusion in EC146(λAZ-7) with both wild-type and mutant ArgRs, β-galactosidase assays were performed. EC146(λAZ-7) does not produce a functional ArgR, and as a result, expresses β-galactosidase constitutively from the argA∷lacZ promoter fusion (128.15 Miller units). In the presence of a wild-type argR gene (present in a pUC19 plasmid), the levels of this enzyme were

reduced 25-fold (3.5 Miller units). A cloned ArgR mutant containing the C-terminal pentapeptide insertion (ArgR5aa) repressed the fusion sevenfold (19 Miller units), and the clone containing the truncated ArgR (ArgR149) repressed 33-fold (5.4 Miller Units) (Fig. 2). The variant ArgR proteins (ArgR5aa and Dabrafenib ArgR149) were then analysed for specific binding to ARG box sites using gel-mobility shift assays. The mutant proteins all retarded the migration of a digoxygenin-labelled E. coli ARG box (Fig. 3). Lanes 2–6 and 9–13 show the effect of the increasing

Liothyronine Sodium concentrations of mutant proteins on their binding activity in the presence of a constant quantity of poly-dIdC and digoxygenin-labelled DNA. A retarded complex was observed at low protein concentrations, which became more apparent as the protein concentration increased. The retarded complexes obtained with the mutant proteins displayed a slightly slower migration than that observed with wild-type ArgR–DNA complexes (Fig. 3, lanes 7 and 14). A labelled nonspecific DNA fragment was not retarded in its migration in the presence of wild-type or mutant ArgR proteins (data not shown). The wild-type and mutant forms of ArgR were then subjected to crosslinking analysis (Fig. 5) using glutaraldehyde. All forms of the protein were able to form higher-order multimeric complexes. Both wild-type ArgR and ArgR5aa form hexamers in the presence of 0.08% glutaraldehyde (Fig. 5, lanes 4 and 8).

Unfortunately, combined influences of maternal TB and co-existing undernutrition are not explored systematically in clinical studies. The potential role of socioeconomic factors55 and maternal impoverished nutrition56 has been suggested in earlier studies from developed

countries. A recent study from India also showed that multiparity, anemia, undernutrition and overcrowding, all added to the problem of maternal TB.10 The risk factors for TB also adversely affect perinatal outcome. Selleck RG7422 It is very difficult, if not impossible to dismantle the potential effects of those risk factors on pregnancy outcome from that of TB. In addition, TB being a chronic debilitating disease requiring long-term care and medication, often consumes enormous financial and non-financial resources

of the family. Furthermore, simultaneously attending maternity care and the TB clinic can be a very daunting task for an indigent family. As a consequence, irregular treatment and advanced tuberculous disease can adversely affect both maternal and perinatal health and survival, especially for women in South Asian countries and ethnic minorities in the UK.7,8,14 Therefore, it is important to consider the problem of TB not as a medical problem alone, but to consider it holistically in the context of socioeconomic background (Fig. 1).57 Anti-TB GDC-0199 mouse drug therapy is only a part of the solution to a more complex issue with medical-social-economic-cultural factors, which need a multidimensional approach ifenprodil from several agencies. Education and emotional support of the affected women and their family members emphasizing the twin need of TB treatment and pregnancy care are two vital issues,29 which can affect successful obstetric outcome. These require the concerted efforts of the public health system and maternity service, which remain suboptimal in most South Asian countries.27 Advocacy, communication and social mobilization are three key factors, which can effectively bridge pre-existing gaps between the health system and the community by enhancing TB knowledge, attitude

and practice.58 It is a sad irony that despite TB largely affecting young women of reproductive age, only piecemeal information about its effects in pregnancy is available, and this incomplete knowledge has clouded our understanding regarding management of TB in pregnant women, and its effect on perinatal outcomes. TB and HIV are inextricably related.23,59 The negative impacts of each on the other have been widely documented.59–62 Both infections occur in women of the reproductive age group.60 HIV infection and TB during pregnancy are considered a ‘deadly combination’ and are independent risk factors for maternal mortality.63 Although Africa is worst affected by this dual disease,23,59 HIV co-infection affects approximately 4–5% of all TB incidence cases in India in 2008 and to a lesser extent, other South Asian countries.

We then combined data sets. Risk quintiles were generated for the HIV biomarker and the combined models. JNK activity Each Poisson model (HIV, ‘non-HIV’, and combined) was used to generate a risk estimate for each subject. Using each set of model estimates in turn, subjects were ranked from highest to lowest risk and grouped into five quintiles designated by equal numbers of mortality events to ensure similar power to detect differences in risk. Observed mortality rates and 95% CIs were estimated. To determine the effect of differing survival intervals on its discrimination, we reran the Index in both development and validation samples censoring survival follow-up at 30 days, 6 months,

1 year, 2 years, 4 years, and 6 years in development and validation selleck chemicals llc samples. For each model, we calculated a C statistic and compared this with published C statistics (receiver operator characteristic area estimates) for two commonly used prognostic indices, Acute Physiology and Chronic Health Evaluation (APACHE) [36] and The Charlson Comorbidity Index [37]. We fitted a logistic model predicting missing data (0 if no data missing and 1 if at least one variable missing) and including all variables (HIV, ‘non-HIV’, substance abuse or dependence, age, mortality, and year of cART initiation). We used predictions from this model to inversely weigh observations in the development and validation sets and compared

these results with those of the complete case analyses. Of 13 586 HIV-infected veterans initiating cART between 1 January 1997 and 1 August 2002 with laboratory data, 9784 (72%) had complete data (analytic sample). Development and validation sets were clinically similar. Subjects were middle-aged (Table 1; median age 45 years), mainly male (98%), and predominantly black (51%). Over a third had CD4 counts below 200 cells/μL and 18% had HIV RNA above 5 log copies/mL. Diagnoses of alcohol or drug abuse or dependence were

common (31%), as were anaemia (21%), HBV infection (12%), and HCV infection (43%). Twelve per cent had likely liver fibrosis (FIB 4>3.25). Rutecarpine Two per cent had stage IV renal failure (eGFR<30 mL/min). AIDS diagnoses were relatively uncommon. In pairwise comparisons, CD4 cell count, HIV RNA and AIDS-defining illnesses were strongly associated with haemoglobin, FIB 4, and eGFR <30 mL/min (P<0.0001 for each; data not otherwise shown). In development and validation sets, HIV and ‘non-HIV’ biomarkers were associated with mortality when modelled separately (Table 2). In both sets, ‘non-HIV’ biomarkers, as a group, added discrimination to the HIV model when combined into a single index [C statistic improved from 0.68 to 0.72 in development (P<0.0001) and from 0.71 to 0.77 in validation (P<0.0001)]. In all cases, all biomarkers retained independent associations with mortality after full adjustment. When data sets were combined, and quintiles of risk estimated, the combined index offered improved differentiation of mortality (Fig. 1).