Pujol for advice This work was supported in part by the project

Pujol for advice. This work was supported in part by the project with reference AGL2011-30461-C02-02 by the Ministerio de Ciencia e Innovación (Spain). 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. “
“The Gram-negative anaerobe Dichelobacter nodosus is the causative agent of footrot in sheep. Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent.

Virulent strains have greater twitching motility and secrete proteases that are more thermostable than those secreted by benign strains. We have identified polynucleotide phosphorylase (PNPase) as a putative virulence regulator and have proposed that PNPase expression is modulated by the adjacent integration of genetic elements. In this study, GSK126 manufacturer we compared PNPase activity in three virulent and four benign strains of D. nodosus and found that PNPase activity is lower in virulent learn more strains. We disrupted the pnpA gene in three benign D. nodosus strains and two virulent strains and showed that deletion of the S1 domain of PNPase reduced catalytic activity. In all but one case,

deletion of the PNPase S1 domain had no effect on the thermostability of extracellular proteases. However, this deletion resulted in an increase in twitching motility in benign, but not in virulent strains. Reconstruction of the pnpA gene in two mutant benign strains reduced twitching motility to the parental level. These results support the hypothesis that PNPase is a virulence repressor in benign strains of D. nodosus. Footrot is a mixed bacterial infection Parvulin of the hooves of sheep, goats and deer that leads to lameness. The Gram-negative anaerobic bacterium Dichelobacter nodosus is the principal causative agent (Beveridge, 1941). Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent. The extracellular proteases secreted by virulent

strains are more thermostable than proteases secreted by benign strains (Depiazzi & Richards, 1979). Virulent strains also have greater twitching motility, generated by polar type IV fimbriae, than benign strains (Depiazzi & Richards, 1985), and twitching motility is essential for virulence (Kennan et al., 2001; Han et al., 2008). Comparative analysis of DNA from virulent and benign strains has led to the identification of a series of genetic elements that integrate into the D. nodosus chromosome. These include the intA (Katz et al., 1991, 1992, 1994; Cheetham et al., 1995; Billington et al., 1996), intB (Bloomfield et al., 1997), intC (Bloomfield et al., 1997) and intD elements (Tanjung et al., 2009), each of which contains an integrase gene. A fifth integrated element, the virulence-related locus, vrl (Katz et al., 1991; Haring et al., 1995; Billington et al., 1999), lacks an integrase gene.

According to Buchner (1960), many hatching larvae and nymphs (eg

According to Buchner (1960), many hatching larvae and nymphs (e.g. Regorafenib weevils, stink bugs) own biting mouthparts with which they feed parts of the eggshell during its burst and are thus infected with the bacteria. Larval infection of P. riparius with endosymbionts most probably takes place in the same manner. However, where exactly the endosymbiotic bacteria of P. riparius are located inside the beetles was not resolved in this work and requires further FISH investigations with the novel oligonucleotide probes developed in this study. We thank J. Piel (Kekulé Institute, University of Bonn) for the supply with the ketosynthase-specific primer

pair KS1F/KS1R, H. Rödel (Institute of Animal Physiology, University of Bayreuth) for the kind introduction in sigmaplot 9.0, R. Grotjahn (Institute of Electron Microscopy, University of Bayreuth) for numerous electron-microscopical exposures, E. Helldörfer (Institute of Animal Ecology II, University of Bayreuth) for the creation of scientific figures of Paederus beetles’ anatomy, W. Nowak and I. Nowak for

the provision of several P. riparius specimens and Harold L. Drake for provision of a LINUX-based network for arb. Financial support by the Deutsche Forschungsgemeinschaft (DFG) is gratefully acknowledged (GRAKO 678). “
“Bacteria secrete small signal molecules into the environment that induce self and neighbour gene expression. This phenomenon, termed quorum sensing, allows cooperative Apoptosis inhibitor behaviours that increase the fitness of the group. The best-studied signal molecules are the N-acylhomoserine lactones (AHLs), Montelukast Sodium secreted by a growing number of bacterial species including important pathogen species such as Pseudomonas aeruginosa. These molecules have recently been proposed to have properties other than those of signalling, functioning as iron quelants or antibiotics. As the presence of an acylase capable of inactivating long-chain AHLs in Anabaena sp. PCC7120 could constitute a defence mechanism against these molecules,

in this work we analyse the effects of different AHLs varying in length and substitutions on the growth and nitrogen metabolism of the cyanobacterium Anabaena sp. PCC7120. All the AHLs tested strongly inhibited nitrogen fixation. The inhibition seems to take place at post-transcriptional level, as no effect on heterocyst differentiation or on the expression of nitrogenase was observed. Moreover, N-(3-oxodecanoyl)-l-homoserine lactone (OC10-HSL) showed a specific cytotoxic effect on this cyanobacterium in the presence of a combined nitrogen source, but the mechanism involved seems to be different from that described so far for tetramic acid derivatives of oxo-substituted AHLs. These results suggest a variety of new unexpected activities for AHLs, at least on cyanobacterial populations. The term ‘quorum sensing’ (QS) (Fuqua et al.

This finding was not surprising, as higher expression levels of m

This finding was not surprising, as higher expression levels of mexB and mexD are to be expected in mexR or nfxB mutants (Dumas et al., 2006). The nature of the specific beneficial adaptive mechanisms in the tolerant bacterial population is under investigation in our lab. To gain insight into the effect of inactivation of genes involved in the GO system on selleck chemicals the global gene expression, we studied the transcriptional changes produced by inactivation of the two genes. The inactivation of both mutY and mutM caused significant changes (more than twofold and P-value < 0.05 compared with PAO1) in six genes (Table 4). Remarkable was the up-regulation of pfpI whose product has been shown to have a protective role against

DNA damage caused by the oxidative stress (Rodriguez-Rojas & Blazquez, 2009). This can be considered a compensatory mechanism for the protection of the DNA in PAOMY-Mgm with impaired repair of the DNA oxidative damage. Interestingly, the most significantly down regulated gene was PA5148 involved in iron selleck trafficking. The modified expression of pfpI and PA5148 expression in PAOMY-Mgm compared with PAO1 was confirmed using RT-PCR, which showed up-regulation of pfpI (9 ± 2.3-fold) and down-regulation of PA5148 (4.04 ± 2.7-fold). Complementation of PAOMY-Mgm,

which showed high expression levels of pfpI and low expression levels of PA5148 compared with PAO1 with wild-type mutM or mutY, reduced the level of pfpI up-regulation to 6 ± 2.4-fold and 4.6 ± 2.4-fold, respectively and the level of PA5148 down-regulation to 1.6 ± 0.09-fold and 2.1 ± 0.25-fold, respectively. Pseudomonas aeruginosa, which colonizes and persists within the highly ROS-rich CF airways has to protect itself against the mutagenic effect of ROS and it uses the GO system, consisting of MutT, MutY and MutM to prevent or eliminate the oxidized form of guanine, which is a mutagenic lesion. Homologue proteins are present in other microorganisms as well as in eukaryotic cells. Inactivation

of each of the three genes encoding for the respective proteins led to various degree of increase in the spontaneous MF with mutants in mutY and mutM exhibiting a moderate and weak mutator phenotype (increase in MF < 20 times the MF of PAO1) (Mandsberg et al., Dipeptidyl peptidase 2009; Morero & Argarana, 2009; Sanders et al., 2009). In the present study, we show for the first time that the mutY and mutM double mutant (PAOMY-Mgm) showed a strong mutator phenotype providing evidence for the cooperation of MutM and MutY to prevent mutagenesis in P. aeruginosa, in a similar manner as in E. coli (Michaels et al., 1992; Tajiri et al., 1995). It has been shown that hypermutability plays an important role in the adaptive evolution of P. aeruginosa in the CF lung (Mena et al., 2008), and it has been demonstrated that mutator populations are amplified by hitchhiking with adaptive mutations. The selective pressure exerted by antibiotics plays an important role in the adaptive process of P.

The ghrelin-mimetic drug growth-hormone releasing peptide 6 (GHRP

The ghrelin-mimetic drug growth-hormone releasing peptide 6 (GHRP-6) has been shown to inhibit light-induced cFos expression in the SCN and attenuate a light induced phase shift (Yi et al., 2006; Yi et al., 2008), suggesting that ghrelin can act as a non-photic stimulus to alter the timing of light-signaled behaviour. Therefore, it is not surprising that the absence of ghrelin could alter the timing of activity, especially in LL, where photic Zeitgebers are also absent. In this situation, the absence of ghrelin activity at the GHRS receptor did not have a significant effect on comsummatory behaviour, as the two groups ate the same amount of food and there were no differences

in body weight. One question that must be addressed is the surprising lack of food anticipatory activity in WT mice housed in LL. Indeed, food anticipatory activity has been previously demonstrated in rats housed Cabozantinib clinical trial in LL (Bolles & Stokes, 1965; Edmonds & Adler, 1977a,b; Lamont et al., 2005). In Lamont et al. (2005), no attempt

was made to quantify the amount of anticipatory activity, but certainly overall activity levels were very low after an extended period in LL, as can be seen in the actograms presented in that article. Species differences may selleck compound account for the lack of food anticipatory activity observed in the present study in WT mice. In one study using spiny mice, Acomys cahirinus, wheel-running activity was reduced dramatically in LL compared to LD and only two of the 11 mice studied actually showed entrainment to a restricted feeding schedule under LL, although all 11 had shown significant food anticipatory activity on an LD schedule prior to exposure to

LL (Chabot et al., 2012). In the current experiment, Celecoxib 30 days in LL reduced daily activity levels in WT mice to fewer than 200 wheel revolutions per day, as compared to 600 in KO mice. With such a low level of activity in WT mice, it may simply be difficult to detect food anticipatory activity in these animals. Sampling of brain and peripheral tissues for clock gene protein and RNA at different time points during the temporal feeding period would have demonstrated whether central and peripheral circadian oscillators were entrained to the time of food availability, although the large number of animals required for this type of study was prohibitive. Alternately, a circadian-controlled measurement that is suppressed by light to a lesser degree, such as body temperature, may have been useful in detecting food anticipation in these mice. Regrettably, these data were not collected. Together, these data provide further support for the hypothesis that ghrelin plays a role in the food-entrainable clock, but also suggest that there may be an interaction between the effect of light and ghrelin that extends beyond a simple deficit in the ability of GHSR-KO animals to entrain to scheduled feeding.

Further research is required to explore the detailed mechanisms

Further research is required to explore the detailed mechanisms. “
“l-Asparaginase-producing microbes are conventionally screened on phenol red l-asparagine-containing plates. However, sometimes the contrast of the zone obtained (between yellow and pink) is not very sharp and distinct. In the present investigation, an improved method for screening of the microorganisms producing extracellular l-asparaginase is reported wherein bromothymol blue (BTB) is incorporated as pH indicator in l-asparagine-containing medium instead of phenol red. Plates containing BTB at acidic pH are yellow and turn dark blue at alkaline selleck products pH. Thus, a dense dark blue zone is formed around microbial colonies producing l-asparaginase, differentiating between enzyme

producers and non-producers. The present method is more sensitive and accurate than the conventional method for screening of both fungi and bacteria producing extracellular l-asparaginase. Furthermore, BTB gives a transient green colour at neutral pH (7.0) and dark blue colour at higher pH 8.0–9.0, indicating the potency of the microorganism for l-asparaginase production. ERK inhibitors library
“Studies of enterohemorrhagic Escherichia

coli (EHEC) infection mechanisms using mammals require large numbers of animals and are both costly and associated with ethical problems. Here, we evaluated the pathogenic mechanisms of EHEC in the silkworm model. Injection of a clinically isolated EHEC O157:H7 Sakai into either the silkworm hemolymph or intraperitoneal fluid of mice killed the host animals. EHEC O157:H7 Sakai deletion mutants of the rfbE gene, which encodes perosamine synthetase, a monosaccharide

component synthetase of the O-antigen, or deletion mutants of the waaL gene, which encodes O-antigen ligase against the lipid A-core region of lipopolysaccharide (LPS), had attenuated killing ability in both silkworms and mice. Introduction of the rfbE gene or the waaL gene into the respective mutants pheromone restored the killing ability in silkworms. Growth of both mutants was inhibited by a major antimicrobial peptide in the silkworm hemolymph, moricin. The viability of both mutants was decreased in swine serum. The bactericidal effect of swine serum against both mutants was inactivated by heat treatment. These findings suggest that the LPS O-antigen of EHEC O157:H7 plays an important defensive role against antimicrobial factors in the host body fluid and is thus essential to the lethal effects of EHEC in animals. Infectious diseases caused by enterohemorrhagic Escherichia coli (EHEC) O157:H7 are a serious clinical problem and are associated with encephalopathy and nephropathy (Tarr, 1995; Law, 2000). An understanding of the molecular mechanisms of EHEC O157:H7 virulence is important for establishing effective therapeutic strategies. Unlike other E. coli strains, EHEC produces Shiga toxins and hemolysins. Shiga toxins are encoded by the stx1 and stx2 genes on the phage DNA that is integrated into the EHEC genome (Sato et al.

5, lane 12) An extensive mutagenesis of the E coli ArgR was per

5, lane 12). An extensive mutagenesis of the E. coli ArgR was performed in order to determine its precise role in cer site-specific recombination. The ArgR protein binds DNA at ARG boxes localized in promoter regions of several genes of the arginine regulon and at the cer site, which

contains half an ARG box. It is quite likely that the DNA-binding activity of this protein is an important contributor to its role as an accessory factor in cer recombination by bringing two cer sites together. However, ArgR by itself cannot support cer recombination in the presence of the XerCD recombinases; PepA is also required for this reaction. Alén et al. (1997) have demonstrated that PepA and ArgR interact directly with cer, forming a complex in which the accessory sequences of two cer sites are interwrapped approximately three times in a right-handed fashion. Using pentapeptide http://www.selleckchem.com/screening/mapk-library.html scanning mutagenesis, we isolated a series of ArgR mutants that showed an approximate 90% reduction in cer recombination, but were still able to repress an argA∷lacZ fusion effectively in vivo (Figs 1 and 2). The mutant proteins also displayed

sequence-specific DNA-binding activity (Fig. 3). All of Pirfenidone the insertions mapped to the same amino acid, between residues 149 and 150 of ArgR (ArgR5aa). This region corresponds to the C-terminal region of ArgR, at the end of the α6-helix (Fig. 4). In order to show that the observed phenotype was due to the disruption of ArgR and was not caused by the additional five amino acids residues, we constructed an ArgR mutant that was truncated at this region. This protein lacks residues 150–156 (ArgR149), Ribonuclease T1 but displays the same properties as ArgR5aa, namely a significant reduction

in cer site-specific recombination in vivo (Fig. 1b), and the ability to bind to DNA at near wild-type levels in vivo (Fig. 2) and in vitro (Fig. 3). Moreover, we were able to detect the same level of DNA retardation as the wild-type protein, which suggests that both ArgR149 and ArgR5aa bind to DNA as hexamers (Fig. 3). In addition, crosslinking studies have shown that wild-type and mutant proteins are capable of forming higher-order structures in solution, although ArgR149 does not appear to form hexamers as efficiently as either wild-type ArgR or ArgR5aa under the crosslinking conditions used (Fig. 5). It is possible that the small C-terminal deletion in ArgR149 prevents this protein from forming a stable hexameric structure. Similar results have been observed with the α A-crystallin protein, where deletions of the terminal 11 amino acids from the C-terminus significantly decreased the oligomeric size of the protein (Thampi & Abraham, 2003). Despite this, ArgR149 can still bind DNA effectively both in vivo and in vitro; the addition of DNA and l-arginine may allow this mutant to form more stable hexamers under these conditions.

Alternative approaches proposed the utilization of MALDI-TOF for

Alternative approaches proposed the utilization of MALDI-TOF for species identification based on the sodA gene (Hinse et al., 2011). Most of these assays were developed using blood-derived clinical cultures of the SBSEC or restricted to a single species. In contrast to blood samples, raw dairy products were shown to contain a large diversity of different lactococci, enterococci, Streptococcus thermophilus, or Streptococcus

agalactiae (Delbès et al., 2007; Younan & Bornstein, 2007; Franciosi et al., 2009; Giannino et al., 2009; Jans, 2011), which increases the requirements regarding specificity of the primers. Even though the genes groESL or sodA provide improved capability to differentiate RGFP966 solubility dmso between species and subspecies, the 16S rRNA gene is still regarded as the recommended target for the initial identification of novel bacteria for which the higher degree of conservation of the 16S rRNA gene can be of advantage (Glazunova et al., 2009). This gene is one of the most important genotypic markers for bacterial taxonomy (Yarza

et al., 2008), and a large number of 16S rRNA gene sequences are available for downstream comparison and further analysis (Benson et al., 2009). It therefore represents an ideal target for the analysis of complex and I-BET-762 nmr less-studied ecological niches such as the human microbiota (Grice et al., 2008; Liu et al., 2008) or spontaneous food fermentations, e.g., the African dairy environment. The high-density and complex microbial communities in these niches could result in unexpected genetic modifications through horizontal gene transfer (HGT), which was observed for African S. infantarius subsp. infantarius as well as for S. thermophilus and other LAB (Hols et al., 2005; Terminal deoxynucleotidyl transferase Makarova et al., 2006; Jans, 2011).

HGT is affecting nearly all genes within prokaryotic genomes; some genes including the 16S rRNA gene are, however, hypothesized to be less affected by HGT (Jain et al., 1999). Therefore, the objective was to utilize the high conservation of the 16S rRNA gene to develop an identification assay applicable to all species within the SBSEC allowing clear differentiation from other streptococci, enterococci, and lactococci regularly found in the dairy environment. The availability of large sets of nucleotide sequences from all members of the SBSEC including dairy isolates (Jans, 2011) enabled the design of a subsequent RFLP for the discrimination of SBSEC species groups. Furthermore, the primers were designed to work with Sanger sequencing for downstream sequence analysis. The assay was then evaluated against reference strains and isolated species of dairy microbial communities. Bacterial reference strains listed in Table 1 were obtained from the Culture Collection University of Gothenburg (CCUG, Gothenburg, Sweden), the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), and the National Collection of Type Cultures (NCTC, Porton Down, UK).

DMURs comprised 1% of MURs provided in the previous

DMURs comprised 1% of MURs provided in the previous Belnacasan in vivo month; key barriers to provision were not receiving discharge medication summaries, and restrictions on provision to housebound patients/patients in care homes. Community pharmacists identified a clear need for DMURs and want to play a greater part in managing patients’ medicines after discharge Targeted medicines use reviews (MUR) were introduced in late

2011 and included reviews after a patient’s discharge from hospital (DMURs) but to date there are no published studies on this important service. The aims of our study were to investigate: i) community pharmacists’ experiences of, and involvement in, provision of DMURs SB203580 price and ii) pharmacists’ suggestions for service improvement. An online survey of community pharmacists in NHS Airedale, Bradford & Leeds (NHS ABL) was conducted in March 2013. The questionnaire was developed drawing on published research and practice literature. Piloting was conducted with six pharmacists and included review by both community and hospital practitioners. Questions were mostly structured, some invited additional comments. Data were analysed using Survey Monkey online

software. Ethical approval was granted by University of Bradford and NHS research governance approval by NHS ABL. Study information and a link to the online survey was publicised by Community Pharmacy West Yorkshire to the 450 pharmacies

in the area. The survey was open for two weeks from March 14th with a reminder after one week. Twenty-six community pharmacists participated; two thirds worked in pharmacies with five or more branches, three quarters had been qualified for 11 years or longer. Twenty respondents reported providing 643 MURs in the previous months, 76% of which were targeted Amobarbital MURs. Seven DMURs (1.1%) were provided by eight pharmacies. More than two thirds of respondents disagreed that patients were well educated about their medicines on leaving hospital. Not knowing when a patient had been in hospital and discharged was the most frequently cited barrier to greater involvement. Discharge medication summaries (DMS) were rarely received, (0–1 per week by most pharmacists), and mainly for patients discharged with a compliance aid. Patients who are not able to visit the pharmacy (those who are housebound or discharged to nursing homes) were reported as key barriers to DMUR provision. Workload, staffing and motivation were far less frequently cited. In addition to increased communication from hospitals respondents rated receipt of discharge summaries, wider permission to conduct telephone MURs for housebound patients and those in nursing homes, and funding for domiciliary MURs, most highly for service improvement.

Lawson and colleagues reported that based on 3 years of data capt

Lawson and colleagues reported that based on 3 years of data captured by the Quarantine Activity and Reporting System (QARS), vaccine-preventable and tropical diseases are not major causes of death in international travelers Selleck ALK inhibitor arriving in the United States.[4] Because malaria is not a communicable disease spread person-to-person, reports of malaria are not requested by CDC Quarantine Stations. Only deaths that occurred during travel (on a conveyance or at a US port of entry) are requested. Thus, QARS did not capture 12 malaria deaths associated with international travel

reported by the US National Malaria Surveillance System during that same time period.[2] While QARS is capable of collecting travel-related illnesses or deaths, it would not be an effective surveillance system for travel-associated mortality due to malaria. The cause of death for travelers who died during travel or upon returning from travel might be captured on the US Standard Certificate of Death.[8] However, only the travel-associated data recorded on the death certificate relate to fatal travel-related injury. As a result, data on returning travelers who

died as a result of travel-related illness will not be captured systematically by the current version of the US death certificate for inclusion in find more US vital statistics data. The risks related to travel may not even be considered in assigning cause of death, especially if the signs and symptoms of disease were not overtly suggestive of

a specific travel-related illness, such as malaria or rickettsia, whose symptoms may be shared with many other less exotic maladies. While travel-related information is obtained from ill patients who are able to provide it, the value GABA Receptor of a travel history collected by a physician is often limited to its use in diagnosis and treatment. Travel histories collected in a clinical setting for treatment are often not collected at all or are incomplete,[9] which can limit a systematic collection of epidemiologic data related to severe travel-related illnesses. Furthermore, if the patient dies during hospitalization or while seeking treatment, an autopsy may not necessarily be performed, and thus the true cause of death remains a mystery. Autopsy rates in the United States have been steadily declining since the 1970s, with 50% of autopsies now performed on persons whose death was related to an external cause, such as assault, suicide, and accidental poisoning.[10] If a returning traveler (who truly had severe malaria) presented to an emergency department 2 weeks after returning from travel, a diagnosis of renal failure might be made based on creatinine levels.

Usuku et al [33] followed the changes in drug resistance mutatio

Usuku et al. [33] followed the changes in drug resistance mutations in http://www.selleckchem.com/products/gsk1120212-jtp-74057.html a patient receiving HAART. Mutations detected in the plasma were not present or were infrequently present in the proviral DNA.

The discrepancy persisted for more than 3 years. It is important to emphasize that the peripheral blood pool of lymphocytes represents about 2% of the total number of lymphocytes in normal young adult men [34]. Schnuda et al. [35] showed that the small blood lymphocytes recirculate continuously between the peripheral blood and the lymph nodes in the rat, with each cycle having a duration of less than 3 min. In this article, we report the results of a prospective study assessing the prevalence and persistence of HIV-1 drug resistance mutations in proviral DNA from purified CD4 cells compared with those in plasma viral RNA before therapy initiation in treatment-naïve patients. We also evaluated the evolution of HIV-1 drug resistance mutations in proviral DNA before and after therapy initiation, and plasma RNA mutation patterns in patients remaining treatment-naïve. As 95 to 99% of

infected cells are CD4 cells [36], and in order to confirm the utility of resistance testing in provirus, we used direct sequencing of HIV-1 proviral DNA in purified CD4 cells to follow the evolution of drug resistance mutations in treated and untreated patients and compared the findings to those obtained from HIV-1 viral RNA using the ABI 310 Selleck BAY 80-6946 Prism (Applied Biosystems, Foster City, California). We further chose not to use cloning but

direct population sequencing as this is routinely used in clinical settings. Between May 2002 and July 2007, genotypic resistance Chlormezanone testing was performed on cell-free and cell-associated virus from 69 patients who were not receiving treatment (Table 1). The study was approved by the local ethics committee and informed consent was obtained from each patient. HIV-1 seropositive status was confirmed according to accepted methods. The therapeutic histories of all patients were checked by asking specific questions when they signed the informed consent form and by consulting their clinical records. When documented histories were absent, we contacted the physicians responsible for the patients’ care. This confirmed each patient as HIV drug naïve. Checking the therapeutic histories of all patients can be difficult but is important when studying drug mutations in treatment-naïve patients. Virus was successfully sequenced for 63 of the 69 selected individuals at baseline, both in plasma and in cells. Fifty-eight per cent of the patients were European and 42% non-European, mostly from central Africa. Thirty-nine per cent of the sequenced HIV-1 viruses were subtype B.