In no case was any evidence of contamination found. Furthermore, real-time PCR assays showed increases of the mmoX gene (encoding for the large hydroxylase subunit of the sMMO) that very closely corresponded with direct microscopic cell counts. Thus, for the first time, clear conclusive proof for the reality of facultative methanotrophy was provided. Remarkably, M. silvestris displayed higher yields, carbon conversion efficiency, Selleckchem Ku-0059436 and growth rates on acetate than on methane. Specifically,

the growth rate of M. silvestris was 0.053 and 0.033 h−1 on acetate and on methane, respectively, suggesting that acetate may be the preferred growth substrate for this microorganism. Shortly thereafter, another acidophilic methanotroph, Methylocapsa aurea, was also identified that could utilize acetate as the sole growth substrate (maximum OD600 nm=0.3, μ=0.006 h−1). As shown in Table 1, neither larger organic acids (citrate, oxalate, malate) nor any tested sugar (glucose, fructose, maltose) could be used as a sole growth substrate (Dunfield et al.,

2010). In contrast to M. silvestris, however, M. aurea only expresses pMMO. Strain purity was determined via: (1) phase-contrast and electron microscopy of acetate-grown cultures; (2) sequencing of more than 21 16S rRNA gene clones from both acetate- and methane-grown cultures; and (3) streaking onto medium with yeast extract and growing cultures with Vincristine ic50 acetate in the absence of methane. In contrast to M. silvestris, however, M. aurea grew best on methane, with a maximum OD600 nm of 1.2 and μ=0.018 h−1. It is interesting to note that all these facultative methanotrophic species are not only acidophilic, but also members of the Beijerinckiaceae family known to include species with broad substrate

ranges. It could thus be hypothesized that facultative methanotrophy will only thrive in a small subset of acidophilic methanotrophs of this family, in environments where organic acids such as acetate are found primarily in the protonated form due to the prevailing low pH, and are thereby more readily taken up (Axe & Bailey, 1995). Facultative methanotrophy, however, does not extend to all acidophilic fantofarone methanotrophs of the Beijerinckiaceae family. For example, Methylocapsa acidophila cannot grow on multicarbon compounds such as malate, acetate, ethanol, succinate, or pyruvate (Dedysh et al., 2002, 2005; Dunfield et al., 2010). As a result of these findings, more effort has been spent to find other facultative methanotrophs, and in the past year, other acidophilic methanotrophs of the genus Methylocystis (family Methylocystaceae) were found that could grow on either methane or acetate (Belova et al., 2011). Specifically, Methylocystis strain H2s, a mild acidophile (optimal growth pH of 6.0–6.

Schools for students with special needs were excluded. The second stage of sampling comprised selection of 25% students from 6th, 7th, and 8th grades of the previously selected schools. The study population included 4086 students: 2272 from Amman, 1425 from Irbid, and 389 from Al-Karak. Selected students were given copies of the questionnaire prepared for this study with consent forms to Bcl-2 expression be signed by their parents or their legal guardians. Cover letters were also attached to the questionnaires to providing additional information about the aim of this project and asking parents to kindly allow their children to participate. Only those with written consent were

included in the study. The diagnostic criteria of DE for each surface were determined according to Smith and Knight ([19]) Tooth Wear Index (TWI)[19] as modified by Millward et al.[20]. TSA HDAC All surfaces of permanent teeth were examined for loss of enamel surface characteristics and/or exposure

of dentin or pulp. Participants were considered as having DE if they had at least one surface that exhibited signs of DE. Students who exhibited changes in dental structure, such as amelogenesis imperfecta, dentinogenesis imperfecta, hypoplasia, diffuse opacities, white spot lesions, tetracycline staining, and fluorosis, were excluded from the study. Excluded teeth also included partially erupted teeth, teeth with orthodontic bands or brackets, extensive restorations and crowns, fractured teeth, surfaces with composite restorations, and fissure sealants. The clinical examination was conducted with students sitting in an ordinary

chair in their class rooms using daylight Ergoloid supplemented with a head light. Teeth were dried with gauze and, when necessary, cotton rolls were used to remove debris. A full mouth examination for DE was performed using a mirror, and information was recorded on a prepared examination form by a research assistant. All examinations were carried out by a single examiner who was trained and calibrated by a university assistant professor of paediatric dentistry by examining 20 patients aged between 12 and 14 years who attended the Jordan University of Science and Technology dental clinics before the commencement of the study. There was a 98.4 percentage of agreement between the two examiners. To assess intraexaminer reliability during the study period, approximately 300 participants of the total sample were examined twice. Thus, for every 25 students examined, the first two students in that group were re-examined. The kappa value of intraexaminer reliability was 0.98. The study utilized a self-reported questionnaire that was an Arabic version of the questionnaire used in the National Diet and Nutrition Survey in the United Kingdom[21].

, 2011) are critical for each of their corresponding sortase activities. When two other residues (Leu263 and Thr265) in this motif were changed to Ala, the effect on the enzyme activity was minimal (Fig. 3). Regarding whether these two residues are important for sortase activity, there are no mutagenesis data available for comparison in other pilus-related sortases. However, in the nonpilus-related SrtA, the corresponding L181A mutation has modest effect on catalytic efficiency, while T183A mutation resulted in a

1200-fold decrease in kcat relative to wild-type SrtA (Frankel et al., 2007). Because our polymerization assay only indicates the presence or the absence of activity, not the rate of the enzymatic activity, quantitative methods

need to be developed to address the effect of these mutations more precisely. To this end, our dot-blot E7080 results did show that there are less FimP Selleck ERK inhibitor components on the surface of T265A mutant than on the surface of the wild-type strain (Fig. S1). It has been proposed that sortases use a catalytic triad composed of His-Cys-Arg during the catalytic process (Table 3). The His204 residue is most likely the His residue in the catalytic triad. The His 204 residue is located 6 Å from the Cys 266 SG atom (Persson, 2011). The H204A mutation effect is consistent with what has been reported about other histidine residues located in the catalytic triads. For instance, in pilus-related sortases, its counterparts His 131in SrtC-1 of S. pneumoniae and His157 in SrtC1 of Group

B Streptococcus were essential for pilus fiber formation in both organisms (Manzano et al., 2009; Cozzi et al., 2011). In the nonpilus-related SrtA from S. aureus, His120 residue at a similar location in relation to the essential Cys184 residue is also critical for the catalytic process (Ilangovan et al., 2001; Ton-That et al., 2002). However, the catalytic function of this critical histidine residue is still a subject of debate. It is speculated that the His residue, because it is being positively charged, might contribute to the electrostatic environment essential for the catalytic activity (Zong et al., 2004). Although the newly published crystal structure (Persson, 2011) showed that Arg275 is part of the His-Cys-Arg catalytic triad, our results for indicate that this Arginine residue is not important for the SrtC1 activity in A. oris T14V. In contrast, its counterparts Arg202 in SrtC-1 of S. pneumoniae and Arg228 in SrtC1 of Group B Streptococcus are essential for the activity of the corresponding sortases (Manzano et al., 2009; Cozzi et al., 2011). Even in the nonpilus-related sortase SrtA, the Arginine 197 residue was identified to be important for the enzyme’s activity(Frankel et al., 2007). However, in SrtA, the Arg197 residue is 13 amino acids away from the essential Cys194 residue instead of the nine-amino acid distance between the arginine and cysteine residues in the SrtC catalytic triads.

The dosing and safety issues with newer therapies, such as lopinavir/ritonavir, are outlined below. It is therefore suggested that neonatal zidovudine monotherapy remains a reasonable approach for infants born to mothers with a HIV VL <50 HIV RNA copies/mL plasma, even if there is a history of zidovudine resistance. Further investigation of the national cohort data to address this question is under way. Where a low transmission-risk mother (see Section 5: Use of antiretroviral therapy in pregnancy) chooses zidovudine

monotherapy plus PLCS, the infant should receive zidovudine monotherapy [1]. There are two situations where triple combination PEP for neonates is advised: Post-delivery infant-only prophylaxis: mother found to be HIV positive after delivery, which is only effective if given buy STA-9090 within 48–72 h of birth. Detectable maternal viraemia (>50 HIV RNA copies/mL) at delivery, mother may be on HAART or not: delivery before complete viral suppression is achieved (e.g. starting HAART late or delivery premature); viral rebound with or without resistance, with or without poor adherence; unplanned Gefitinib supplier delivery ( e.g. premature delivery

before starting ART or late presentation when maternal HIV parameters may be unknown). 8.1.2 Infants <72 h old, born to untreated HIV-positive mothers, should immediately initiate three-drug ART for 4 weeks. Grading: 1C There is one large RCT of combination therapy in neonates born to mothers who did not receive any ART before delivery (n = 1684, in Brazil, Argentina, South Africa and the USA) [18]. Infants were randomly allocated at <48 h of age to: 6 weeks of zidovudine monotherapy; or 6 weeks of zidovudine with three doses of nevirapine in the first week of life; or 6 weeks of zidovudine, with nelfinavir and lamivudine for 2 weeks. Overall, in this

high-risk group, the HIV transmission rate was 8.5%, and in multivariate analysis, only ART arm and maternal VL were significantly associated with transmission. For infants uninfected at birth, transmission Bay 11-7085 was twofold higher in the zidovudine-alone arm compared to the multiple ART arms (P = 0.034). There was no significant difference in transmission rates between the two multiple ARV arms and neonatal neutropenia was significantly higher in the three-drug arm. In a randomized African study, babies born to mothers presenting at delivery received single-dose nevirapine or single-dose nevirapine and 1 week of zidovudine. Of those HIV negative at birth, 34 (7.7%) who received nevirapine plus zidovudine and 51 (12.1%) who received nevirapine alone were infected (P = 0.03): a protective efficacy of 36% for the dual combination [19]. However, in two other randomized African studies where the mothers received short-course ART, for infants uninfected at birth there was no significant difference in transmission rate at 6 weeks for dual vs. monotherapy short-course regimens to the infant: zidovudine plus lamivudine vs.

We report the results from further analyses investigating the frequency, time distribution and severity of AEs

and laboratory abnormalities of interest for etravirine, performed using the week 96 data set from the DUET trials. For these analyses, AEs of interest were selected based on their relevance in the target population (i.e. treatment-experienced, HIV-1-infected patients), their known association with other antiretrovirals and their potential importance based on preclinical or earlier clinical data. We also present the frequency of AEs and laboratory abnormalities per 100 patient-years of exposure for all AEs and laboratory abnormalities of interest to account buy Metformin for the difference in extent of exposure between the etravirine and placebo groups. DUET-1 (NCT00254046) and DUET-2 (NCT00255099) were randomized, double-blind, placebo-controlled, phase III trials of 48 weeks’ duration, with an optional open-label 48-week extension period. Patients were randomized to receive etravirine 200 mg twice daily (bid) or placebo, both in combination with a background regimen of darunavir/ritonavir 600/100 mg bid, investigator-selected nucleoside

reverse transcriptase inhibitors and optional PI3K activity enfuvirtide. The DUET trial design and methodology have been previously reported in detail [3, 6, 7]. Treatment-experienced, HIV-1-infected patients with plasma viral load > 5000 HIV-1 RNA copies/mL were enrolled if they had been on stable therapy for ≥ 8 weeks, had at least one NNRTI resistance-associated mutation (RAM) and at least three primary PTK6 protease inhibitor mutations at screening or in documented historical genotype. The trial protocols were reviewed and approved by the relevant Independent Ethics Committees or Institutional Review Boards,

and written informed consent was obtained from all participants prior to any trial-related procedures. The trials were conducted according to the principles of good clinical practice, the Declaration of Helsinki and the European Union Clinical Trials Directive. The week 96 pooled analysis of DUET-1 and DUET-2 was pre-specified. Safety assessments were carried out every 8 weeks between week 48 and week 96. For the purposes of this analysis, AEs of interest (and preferred terms) were: nervous system AEs (e.g. headache, dizziness, somnolence, memory impairment, amnesia, disturbance in attention, balance disorder, and restless legs syndrome); psychiatric AEs (e.g. depression, insomnia, anxiety, sleep disorder, libido decreased, abnormal dreams, stress, confusional state, nightmare and panic attack); rash-related AEs (e.g.

Scanning electronic microscopy was conducted using cryo-SEM (Hitachi S-3000N microscope, Japan), operating between 10 and 15 kV on samples containing a thin layer of gold sputter coating. Strain R5-6-1 was cultivated on PDA medium for 5 days in the dark at 25 °C, during which time conidiophore and conidia formation started. The margin of the Selumetinib molecular weight culture was then sliced out. The operation was carried out carefully not

to deform the surface features of the culture. The fungal strain was cultured in PD broth for 4 days at 180 r.p.m. min−1 in an orbital shaker at 25 °C. Fungal DNA was extracted using the Multisource Genomic DNA Miniprep Kit (Axygen Bioscience, Inc.) following the manufacturer’s instructions. Primers ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) (White et al., 1990) were used for amplification of the fungal rDNA internal transcribed spacer (ITS) regions 1 and 2. The PCR reaction (50 μL total volume) contained 5 μL 10 × PCR buffer, 7 μL 25 mM Mg2+,

2 μL 2.5 mM dNTP, 2 μL of each primer (10 μM), 4 μL (0.5–10.0 ng) of total DNA, 1 μL Taq polymerase and 27 μL ddH2O. Thirty-five cycles were run, each consisting of a denaturation step at 94 °C (40 s), an annealing step at 54 °C (50 s) and an extension step at 72 °C (60 s). After the 35th cycle, a final 10-min extension step at 72 °C was performed. The reaction products were separated in a 1.0% w/v agarose gel and bands were stained with ethidium bromide. The PCR products were then purified using the DNA Gel Extraction Wnt antagonist Kit (Axygen Bioscience, Inc.) and sequenced in an ABI 3730 sequencer (Applied Biosystems) using the ITS1 and ITS4 primers. The sequences were subjected to a blast search and were

aligned using clustal x together with the next neighbors (i.e. sequences that had a negative probability e-value of 0.0 in a blast search against the GenBank database); the alignment was manually corrected in genedoc. The evolutionary distance was determined using the Jukes–Cantor model to construct a phylogenetic tree by the neighbor-joining method using phylogeny inference package (phylip, v 3.68). The resultant trees were analyzed using the program consense to calculate a majority rule consensus tree. The tree file was then displayed by treeview. Bootstrap (1000 replicates) analysis used SEOBOOT, DNADIST, NEIGHBOR PI3K inhibitor and CONSENSE in phylip. Sequence inspection of the ITS1, 5.8S rRNA gene and ITS2 regions showed 100% identity of H. oryzae isolates R5-6-1 and RC-3-1. The blast similarity search revealed that H. oryzae shared 96%, 95% and 95% identity with ITS 1 and 2 sequences of unidentified Harpophora spp. (AJ132541), Harpophora spp. (AJ132542) and Harpophora spp. (AJ010039), respectively. In order to relate H. oryzae to already known Harpophora sequences and species and other related genera in Magnaporthaceae, a phylogenetic analysis was performed. As shown in Fig. 1, the NJ tree grouped Harpophora spp.

Taking life-long treatment with a high adherence demand may also have emotional effects. Some compounds exacerbate mental health symptoms [7], while others may be associated with side effects (e.g. lipodystrophy) with mental health sequelae [8]. Poor mental health or heavy mental health burden is associated with reduced adherence, which in turn is associated with poorer outcome [6-9]. Therefore, incorporating assessment of mental health

into the routine follow-up of patients at all stages is important but is particularly critical at first presentation in order to establish a baseline. It is also important prior to commencement of ART (see 6.2 Monitoring of ART-naïve patients) and in those individuals with suboptimal adherence and/or virological failure, or signs of mental health symptoms (such DAPT ic50 as depressed mood, heightened anxiety, relationship concerns, memory or functioning concerns). Cognitive symptoms have been noted from the early days of the

epidemic, ranging from mild cognitive symptoms to more severe memory loss, executive functioning difficulties and cognitive impairment [10]. The advent of treatment has clearly reduced the prevalence of severe cognitive disorders [11, 12], while milder forms have continued in a proportion of patients. There is currently much debate about the prevalence, risk factors for, and prognosis of, mild-to-moderate cognitive impairment in persons taking effective ART GSK2118436 purchase (full virological suppression). Joint psychological support standards are currently being consulted on and it is anticipated that these will make recommendations about screening [13], although there is not yet consensus about easy-to-administer and effective measurements. The finalized standards will be available late in 2011. Standardized monitoring of psychological wellbeing at baseline, at annual follow-up and at change points (such as treatment initiation and treatment switching) (III). Having good referral mechanisms to psychological services in place and clear criteria for referral (see BHIVA guidelines on psychological support

[13]) (IV). Inclusion of psychological selleck kinase inhibitor consideration in relation to fertility, drug use, treatment change, side effects, adherence, relationships and doctor–patient interaction (IV). There is no high-grade evidence for what is the optimal frequency at which to measure CD4 T cells in well-resourced health environments. We have considered three different scenarios: initial HIV diagnosis; monitoring ART-naïve patients; and CD4 T-cell counts in patients on ART. Recommendations for how often we should be measuring CD4 T-cell counts are mainly based on expert opinion [1-3]. For ART-naïve patients, we used data from a cost-effectiveness analysis using an HIV simulation model incorporating CD4 T-cell count and plasma HIV-1 RNA load as predictors of disease progression [4].

No transmissions were observed in the UK and Ireland among the 464 pregnancies managed by zidovudine monotherapy and PLCS between 2000 and 2006 reported to the NSHPC. The median delivery VL in these women was 400 (IQR 61–1992) HIV RNA copies/mL [1]. 5.3.5 Women who do not require treatment

for themselves should commence temporary HAART at the start of the second trimester if the baseline VL is >30 000 HIV RNA copies/mL plasma. (Consider starting earlier if VL > 100 000 HIV RNA copies/mL.) Grading: 1C VL data also influence recommendations relating to mode of delivery (see below). Major determinants of the probability of achieving a VL <50 HIV RNA copies/mL plasma by the time of delivery are the baseline untreated VL and the time available to achieve this target. In the Mma Bana Alpelisib order study, VLs <400 HIV RNA copies/mL plasma were achieved by the time of delivery in 96% (lopinavir/ritonavir-based) to 100% (abacavir/lamivudine/zidovudine) of mothers with baseline VL <1000 HIV

RNA copies/mL plasma and in 86% (lopinavir/ritonavir-based) to 90% (abacavir/lamivudine/zidovudine) if baseline VL >100 000 HIV RNA copies/mL. When therapy was initiated at 31–34 weeks, only 78% of mothers on PI-based therapy had achieved this target [21]. Data from a UK multicentre study retrospectively analysing therapy outcomes in pregnant women initiating HAART at a median gestation of 23 weeks demonstrate very Selleckchem Gemcitabine low rates of complete suppression in women with a baseline VL in the upper quartile (>32 641 HIV RNA copies/mL) with only 46% achieving <50 HIV RNA copies/mL by 36 weeks' gestation (the data point used to make most delivery management decisions) and this fell to 37% for VLs >100 000 HIV RNA copies/mL [88]. For all VLs >10 000 HIV RNA

copies/mL, treatment initiation later than 20.3 weeks’ gestation was associated with significantly less likelihood of successful VL suppression. To address this, the Writing Group recommend that HAART should be commenced at the start of the second trimester, or as soon as possible thereafter, in women with a baseline VL >30 000 HIV RNA copies/mL plasma. 5.4.1 A woman who presents after 28 weeks should commence HAART without delay. Grading: Fossariinae 1B Late presentation after 28 weeks and before the onset of labour occurs less frequently since the introduction of the routine offer and recommendation of antenatal HIV screening. With improved turnaround times for VL testing, a woman presenting beyond 28 weeks may still be managed with a view to a possible vaginal delivery if she commences HAART and achieves a VL <50 HIV RNA copies/mL by 36 weeks. Where women present between 24 and 28 weeks, the advantages of more detailed assessment and tailoring of the regimen should be weighed against the advantages of initiating HAART immediately. The turnaround time for CD4 cell counts, VL and viral resistance tests will impact on this choice. 5.4.

Between 24% and 41% of the respondents indicated their inability to determine the appropriate treatment modality for children with high and low caries risk. Majority of the students failed to differentiate between the caries-preventive practice for children with high and low risk of caries: preventive strategies for children with high caries risk were also

used for those with low caries risk. Age, gender, knowledge of caries prevention measures, and self-perceived competency in providing caries-preventive care were not associated with student’s capacity to provide caries-preventive practice for children. Dasatinib mouse Caries-preventive practice among dental students in Nigeria could be improved. It may be important to explore the possible role of problem-based learning selleck monoclonal humanized antibody inhibitor approach in addressing this challenge. Dental

caries has been defined as ‘localized destruction of the tooth surface initiated by decalcification of the enamel followed by enzymatic lysis of organic structures and leading to cavity formation’[1]. Although the disease is not life threatening, it is a matter of great concern in dental public health circles because of its high prevalence in some of the developing countries[2, 3], its consequences such as pain and dysfunction, its impacts on the quality of life at all ages, and its social and economic burdens[4, 5]. The burden of caries is high among children living in Nigeria. Unfortunately, there is only one national study on the caries in children in Nigeria and this was conducted in 1995. The survey showed that 30% and 43% of 12- and 15-year-old children had caries. The DMFT for the 12- and 15- year-olds was 0.7 and 1.2 respectively, with very low level of restorative care[6]. The multiple regional studies in the very country continue to show that the prevalence of dental caries in the permanent dentition remains high ranging from 13.9% in Ile-Ife to 33.0% in Benin, 15.5% to 35.5% in Enugu, 5.7% to 30.8% in Lagos, and 11.2% in Ibadan[7-16] (O. O. Sofola, M. O. Folayan, A. B. Oginni, personal communication). In the primary dentition, the prevalence ranges from 10.9% in Ile-Ife to 6.4%

to 22.5% in Lagos[7, 8, 11, 15-18]. Not only is the prevalence high, the level of untreated caries in the permanent dentition is also high, ranging from 77.2% in Ile-Ife to 98.6% in Benin, and 49.5% to 85.5% in Enugu[7, 12-14]. In Lagos, the restorative index ranged from 0.3% to 1%[8, 16], whereas the treatment index is 5.7%[7]. The Met Need Index in Ibadan was 0.11[8]. The prevalence of untreated caries in the primary dentition also remained high ranging from 92% in Ile-Ife to 95.6% in Lagos[3, 7, 16, 18]. As implementation of a curative and restorative approach to combat dental caries at the population level does not appear to be cost-effective in many countries[5], the World Health Organization (WHO) has put more emphasis on prevention in setting global oral health goals for the year 2020[19].

Among the resistance genotyping tests performed in two hospitals in Paris, France during the last 6 years, either for an indication of virological failure or for an indication of initial diagnosis of HIV infection, we identified cases of virus exhibiting protease gene insertions, and retrospectively collected therapeutic, immunological

and virological data. The proportion of patients infected with HIV-1 non-B subtypes in the two hospitals was 39.9% (including Gefitinib 2.9% CRF01_AE, 22.6% CRF02_AG and 1.2% G). GRT was performed on samples available before and/or after the initial detection of a protease insertion. GRT was performed using the consensus technique developed by the Agence Nationale de Recherche sur le SIDA (ANRS) Resistance Study Group, as previously described [14]. The mutations reported in this study are given in the 2008 International AIDS Society (IAS-USA) list [15]. In order to assess the archiving of the insert-containing virus in the cellular reservoir, GRT was performed on HIV DNA obtained from peripheral blood mononuclear cell (PBMC) specimens when HIV-1 RNA plasma viral load was undetectable, at two different time-points in patient 1 and at one time-point Selleck Cabozantinib in patient 4. Phenotypic resistance

to PIs was determined using the HIV-Phenoscript® PI assay (Eurofins, Kalamazoo, MI) as previously described [16,17]. The gag-protease fragment includes cleavage sites p24/p2, p2/p7, p7/p1 and p1/p6. Furthermore, to assess the replicative capacity of different primary viruses, the region spanning the gag cleavage sites as well as the protease and part of the RT were amplified [18]. The results of the assay are expressed as the sensitivity fold change (FC) 50% inhibitory concentration (IC50) values and as the percentage of replicative capacity

compared with the control wild-type virus (NL4-3). All available PIs, except darunavir (DRV), were tested: amprenavir (APV), atazanavir (ATV), indinavir (IDV), lopinavir (LPV), nelfinavir (NFV), saquinavir (SQV) Pyruvate dehydrogenase lipoamide kinase isozyme 1 and tipranavir (TPV). Eleven patients were found to harbour plasma virus with a protease insertion, giving a frequency of 0.24% (11 of 4500 patients). Two patients were ARV-naïve, one was PI-naïve and eight were PI-experienced (Table 1). The inserts were composed of one or two amino acids which mapped between codons 33 and 39 for 10 patients and at codon 19 for one patient (Table 1). The nucleic acid composition of the inserts mainly consisted of duplications of neighbouring sequences (Table 2). At the time of detection, the insertion-containing virus had a median of 9 mutations associated with PI resistance (range 3–13). Six patients (55%) were infected with a HIV-1 non-B subtype (three with CRF02_AG, one with CRF01_AE, one with subtype A and one with subtype G) and most of the mutations were subtype-specific polymorphisms, as confirmed by the Stanford database (http://hivdb.stanford.edu/cgi-bin/MutPrevBySubtypeRx.cgi) (Table 1).