Gudger analyzed these accounts, but he still remained skeptical overall. Yet,

he listed the names of eight men whom he could accept as eye witnesses, admitting that just because something seems improbable does not mean it does not exist. Reexamining the material for this paper, the various accounts, especially original documents (de Castelnau,[9] von den Steinen,[11, 12] Pellegrin,[13] Jobert,[21] and Boulenger[22]), illustrate that most reports are, in fact, repeated again and again based on the same stories already described elsewhere. Therefore, after careful distillation, very little remains and of that little, even accounts sounding like first-hand descriptions become suspect. H.H. Rusby had claimed that “evidence is abundant and confirmed,” but he failed to provide proof.[16] In retrospect, it is almost impossible to identify genuine eye witnesses of candiru “attacks” and we just have to trust Cabozantinib that some reports may, indeed, be true. A number of critical comments shall be made here, not only because it is important

to http://www.selleckchem.com/products/azd9291.html interpret the literature mindfully but because it is the basis of current medical advice. These comments relate to the exoticism of the topic, local language issues, and the translation of original accounts. Modern travel, even to the most remote places, has no parallel in early voyages. It is difficult today to appreciate fully the physical and mental challenges these explorers faced. Devoted to their particular field of interest, they traveled through unknown, often hostile, environments, collecting astonishing objects and information along the way. Something as bizarre as a fish swimming up people’s urethra must have been one of the most exhilarating stories of the time. Of adventurous spirit and in exotic surroundings, it is easy to get carried away. In such circumstances, a first report, relayed with caution, can quickly take

on a life of its own and, embellished with more and more gruesome details, eventually becomes a fact. It would have taken little to keep the stories alive. The smallest rumor, added to the “body of knowledge,” simply confirmed now preconceived expectations. On the other hand, despite their captivating accounts, it appears that many explorers’ verdict remains one of skepticism because of the absence of scientific proof. Another Methocarbamol point of caution is the use of local languages in obtaining reports from indigenous tribes. Some explorers studied local languages and would have been able to converse with local informants to some degree. However, others and those who traveled for long periods of time and over considerable distances would not have been in a position to speak all the languages encountered. Despite the use of língua geral,[23] a unifying language based on Old Tupi, there is still a great potential for misinterpretation of language, postures, and gestures.

Furthermore, the term ‘adverse drug event’ was used as a medication error search term. This returned over 10 000 additional results. The first 300 articles were related to the harm due to drug use. However, this review aimed to identify failures in the medication use process in order to provide an overview of the overall reliability, efficiency and safety. The search strategy, tailored for each database, therefore included two concepts, medication error and primary care,

and excluded a third, secondary care (Table 1). ‘Medication error’ was used as MeSH term and keyword. A hand search of Belinostat supplier key journals, which included International Journal of Pharmacy Practice (IJPP), Quality and Safety in Healthcare and Pharmacy World and Science, was also performed. Studies conducted in any country between January 1999 and November 2012 and reported in English were included. Studies, which reported the frequency of errors in the medicines management process, and interventions to prevent errors, were included. All definitions of error such as inappropriate prescribing; prescribing, dispensing, administration and monitoring errors; irrational drug use; hazardous prescribing; and drug interactions

were included. Studies estimating error rates of one medication or therapeutic group, and those that did not report the method used for collecting error data or evaluating interventions, were excluded. find more The first author (JOO) screened all titles and abstracts to determine whether Mirabegron the article met the inclusion criteria and should be retrieved. Another reviewer (MG)

screened a random 5% sample to check the reliability of the screening. JOO then read and extracted data from the articles included in this review. Search results were exported to Endnote X5 (Thomson Reuters, Times Square, New York, NY, USA). Duplicates were removed. Article titles and abstracts were initially reviewed for relevance followed by actual article review to clarify any ambiguities. Information from incidence studies was extracted onto a pro-forma showing primary author, year of publication, study design and setting, sample size, error type, error definitions and reported error rates (Table 2a). Intervention studies were grouped into broad categories (Table 3). Near miss’ incident that was detected up to, including the point at which medication was handed over to patient or their representative’ Incidents detected after patients had taken possession of medication were recorded as ‘dispensing errors The output of the search process is shown in Figure 1. Thirty-two studies, which estimated the incidence of medication errors in primary care, were identified; a manual search retrieved one additional study.[19] Thus, 33 studies were identified and reviewed (Table 2b).

This study is among the largest cohort studies on HIV nPEP. It includes a population of subjects potentially exposed to HIV through various routes, both sexual and nonsexual. Our results demonstrate the

feasibility and efficiency of a strategy based on active tracing of the source of exposure as a means to reduce unnecessary antiretroviral prophylaxis. Current CDC guidelines recommend the prescription of nPEP in cases of exposure to a known HIV-infected source [7]. A study by Pinkerton et al. [23] concluded that nPEP was only cost-effective in cases where men reported receptive anal intercourse with an infected partner. ZD1839 However, HIV transmission by partners of unknown HIV status has already been reported in this context [24]. In cases of nonoccupational exposures, especially for anonymous sexual contacts, the HIV status of the source is often unknown, as was the case in our study for 77% of events. CDC guidelines do not recommend for or against the use of nPEP in these situations but favour a case-by-case approach in which risks and benefits are weighed [7]. Swiss national guidelines recommend prophylaxis in situations where the source person belongs to a high-risk group for HIV infection (MSM, IDU, individuals from high HIV prevalence areas and sexual assaulters)

[15]. For this reason, in most nPEP studies published to date, antiretroviral prophylaxis has been provided for both documented and high-risk selleck chemical potential exposures to HIV [12,13,16–20]. The only way to overcome this problem and avoid unnecessary prescription of antiviral prophylaxis is to test the source subject whenever possible, as stressed by some guidelines [7,25]. Tracing and testing the source person has already proved feasible and cost-saving [20,26]. In a previous report based on a smaller sample of the same cohort, this strategy was found to reduce the number

of nPEP prescriptions by 28% [26]. In our study, source persons of unknown HIV status could be tested in 42% of events, a proportion significantly higher than previously reported (7–16%) [16,20]. The reason why we obtained such a high rate of source persons presenting for testing was probably related to the proactive way in which we explained to the exposed patients the benefits of avoiding Cell press or interrupting nPEP if the source was tested negative for HIV. These included not having to be exposed to antiretroviral drugs with known side effects for 28 days and the financial benefit of not paying for the entire course of nPEP (in Switzerland, the cost of nPEP is charged directly to the patient and then partially reimbursed through medical insurance). This approach allowed us to avoid or interrupt unnecessary nPEP in 31% of eligible events, contributing to reduced healthcare costs, potential drug toxicity and anxiety for the exposed person.

Coxiella burnetii NMII infections were initiated as described and fixed at 0, 8, 16, 24, 48, 96, and 168 hpi with 4% paraformaldehyde, 0.05% Tween-20 in phosphate-buffered saline for 15 min at room temperature. Indirect immunofluorescent antibody (IFA) analysis was performed by dual staining as described previously (Morgan et al., 2010). Micrograph images were captured via a Nikon DS FI1 camera on a Nikon Eclipse TE 2000-S microscope at × 400 magnification, with nis-elements f 3.00 software. A magnification of × 400 was used as opposed to × 600, as used previously (Morgan et al., 2010). Micrograph capture settings were uniform for all images. Using a modification of a method used previously in C.

burnetii studies that uses relative pixel ratios in sample quantitation (Zamboni et al., 2001), each micrograph image was analyzed using imagej version EPZ015666 manufacturer 1.42n (Wayne Rasband, NIH) software. Five fields of view from each time sampled (three biological samples of each) were digitally captured. The matching Alexa Fluor® 555 and Alexa Fluor® 488 images were stacked (paired) and converted to gray scale (8 bit). No fewer than five regions of interest (ROI) were blindly selected from each field of view of the 555 nm grayscale images. This provided at least 75 ROIs for each time point analyzed. Saturated regions of an image were not selected and ROI size varied

BIBF 1120 molecular weight depending on the PV size. The pixel densities within the identical ROIs from each stacked image were then measured

as published previously (Collins, 2007). The mean pixel densities were then compared to obtain the 488 : 555 ratio for each ROI. These individual ratios were then averaged (≥75 individual ratios/time point) to determine the relative expression of IcmT to whole C. burnetii NMII. The final 488 : 555 (IcmT : C. Ureohydrolase burnetii) ratio for each time point was then divided into the 0 hpi ratio to obtain the final IcmT relative expression levels. The statistical significance between each time point was evaluated using single-factor anova with a 95% confidence interval using ms excel 2007 (Microsoft). The C. burnetii T4BSS RI gene linkage map suggests that three groups of transcriptionally linked genes exist (see Fig. 1a). These include: (1) icmXCBU1651icmW, (2) icmVdotACBU1647, and (3) CBU1646dotBdotCdotDicmSicmT. To demonstrate transcriptional linkage between the genes within each group, RT-PCR analysis was performed using oligonucleotide primers (see Table 1) designed to span intergenic sequences and/or adjoining ORFs. The diamond-ended lines in Fig. 1a indicate the position of primers and DNA products that would result from RT-PCR amplification. Using total RNA harvested from Vero cells infected with C. burnetii NMII as a template, amplification products were observed (Fig. 1b) for each linkage region: (1) icmW–icmX, (2) icmV–dotA, dotA–CBU1647, and (3) icmT–dotD, dotD–dotB, dotB–CBU1646 (Fig. 1b).

If such analogues could be readily produced and were sufficiently stable for clinical use, then renewed attempts to develop

further derivatives of PAS would appear worthwhile (e.g. see Patole et al., 2006). The dual administration of two inhibitors, one preventing the synthesis of salicylate and the other stopping its conversion to mycobactin, could therefore be an extremely effective way of preventing the growth of mycobacteria and could therefore be useful in the treatment of tuberculosis. We thank Overseas Research Studentships (UK) for a research studentship to N.N. We also thank Dr Andrew Boa, Department of Chemistry, University of Hull, UK, for helpful discussions. “
“Autophagy is a degradation system in which cellular components Epacadostat are digested via vacuoles/lysosomes. APO866 cost In the budding yeast Saccharomyces cerevisiae, the induction of autophagy results from inactivation of target of rapamycin complex 1 (TORC1), promoting formation of the serine/threonine kinase Atg1, which is one of the key autophagy-related (Atg) proteins required for both nonselective and selective autophagy such as the cytoplasm-to-vacuole targeting (Cvt) pathway. Here, to understand the induction mechanism of autophagy in filamentous fungi, we first identified the ATG1 homolog Aoatg1 in Aspergillus oryzae and then analyzed the localization

of an enhanced green fluorescent protein (EGFP)–AoAtg1 fusion protein. AoAtg1–EGFP localized to pre-autophagosomal structure (PAS)-like structures, similar to Atg1 localization

in S. cerevisiae. The function of AoAtg1 was evaluated by constructing an Aoatg1 disruptant, ΔAoatg1. Conidiation and development of aerial hyphae were scarcely observed in ΔAoatg1. Moreover, autophagy in the disruptant was examined by observation of the localization of EGFP–AoAtg8 and AoApe1–EGFP, with the results indicating that AoAtg1 Tolmetin is essential for nonselective autophagy and the Cvt pathway. Furthermore, we demonstrated that the overexpression of Aoatg1 results in decreased conidiation and the excessive development of aerial hyphae and sclerotia. Taken together, our findings provide evidence for the existence of the Cvt pathway in A. oryzae. Macroautophagy (hereafter autophagy) is a highly conserved degradation pathway that mediates the turnover of bulk cytoplasmic protein and organelles induced under nutritional starvation conditions (Nakatogawa et al., 2009). Autophagy plays a number of roles associated with quality and quantity control of cytoplasmic components, including the killing of intracellular microorganisms (Deretic & Levine, 2009) and removal of damaged or depolarized mitochondria (Apostolova et al., 2011). The autophagic process consists of several sequential steps: the induction of autophagy, autophagosome formation, fusion of autophagosomes to lysosomes/vacuoles, and degradation of autophagic bodies (Mizushima, 2007).

All were obtained from the same reputable supplier at different dates, and paired with primer R907 (Teske et al., 1996). Soil community PCR was performed with

a 25-μL reaction mixture containing Protein Tyrosine Kinase inhibitor 1.25 U GoTaq polymerase (Promega), 1 × PCR buffer, 1.5 mM MgCl2, 0.4 mg mL−1 bovine serum albumin, 4 μM each primer, 200 μM dNTPs, and 10 ng of template DNA. Pure-culture PCR was performed using a 30-μL reaction mix using the above concentrations of reagents. Thermocycling conditions were as follows: initial denaturation at 94 °C for 5 min, followed by 35 cycles of 94 °C for 30 s, 55 °C for 45 s, and 72 °C for 1 min, and a final elongation at 72 °C for 7 min. After PCR, 1 μL of PCR product was resolved in a 1.5% agarose gel to confirm product size and the negative control. DNA concentrations were determined throughout by fluorometry using the HS dsDNA kit and Qubit Fluorometer (Invitrogen). A total of 300 ng of PCR product was loaded into each lane for soil community DGGE, while 50 ng of DNA was loaded for pure-culture DGGE. A denaturing gradient of 35–65% denaturants [100% denaturants is a mixture of 7 M urea and 40% (v/v) formamide] (Muyzer et al., 1993) was used in 6% (w/v) polyacrylamide gels. Electrophoresis was performed in 1 × Tris-acetate EDTA buffer at 60 °C

and at a constant voltage of 70 V for 16 h using a DCode system (BioRad). The DGGE gels were stained in a 1 : 2000-diluted SybrGold (Molecular Probes) in water Ribociclib datasheet Florfenicol for 30 min. Gel images were captured

using a ChemiDoc XRS (BioRad), and analyzed using quantity one software (BioRad). The background was subtracted using a rolling disk set at 20, and band density at positions was converted to intensity per Rf value between 0 and 1. After normalizing for total intensity across lanes, data were input into the past software package and analyzed using multivariate principal component analysis (Hammer et al., 2001). PCR product from each primer set was ligated into pGEM-T Easy Vector (Promega) and transformed into E. coli JM109 competent cells. A total of 10 clones from each primer set reaction were chosen at random for sequencing. The DNA sequences were determined using the chain termination method by the Nevada Genomics Center, using the T7 primer. Vector sequence and 16S rRNA gene sequences downstream of the respective primer were removed manually. The melting temperature (Tm) was calculated with biomatch (Promega), using the base-stacking algorithm. Empirical observations of differences in DGGE profiles generated using separate GC-clamp primer batches lead us to suspect variation in performance of distinct batches. Therefore, we PCR-amplified two soil DNA extracts using three batches (N1–N3) of the same GC-clamp primer, paired with the same reverse primer R907. To compare the effect of a longer template-directed sequence, primer G1 had the same GC-clamp sequence but an 18- rather than a 15-base 16S rRNA gene sequence (Muyzer et al., 1993).

and Sphingobium sp. The results show that aerobic microbial granules based process is suitable for the treatment selleck of dibutyl phosphite contaminated water. “
“This work describes an efficient, simple, and green bioprocess for obtaining 5-halogenated pyrimidine nucleosides from thymidine by transglycosylation using whole cells. Biosynthesis of 5-fluoro-2′-deoxyuridine (floxuridine) was achieved by free and immobilized Aeromonas salmonicida ATCC 27013 with an 80% and 65% conversion occurring

in 1 h, respectively. The immobilized biocatalyst was stable for more than 4 months in storage conditions (4 °C) and could be reused at least 30 times without loss of its activity. This microorganism was able to biosynthesize 2.0 mg L−1 min−1 (60%) of 5-chloro-2′-deoxyuridine in 3 h. These halogenated pyrimidine 2′-deoxynucleosides are used as antitumoral agents. Nucleosides have been considered of great interest because they have shown activity against various cancer

cell lines in vitro and in vivo. Nucleosides and their analogues are implicated in the modulation of several signal transduction pathways causing growth inhibition, differentiation, apoptosis, and modulation of gene expression through different mechanisms of action (Wang et al., 2004; Rossi et al., 2009; Li et al., 2010). Therefore, nucleoside analogues can be used as powerful antitumoral agents. Halogenated derivatives are high throughput screening compounds widely recognized today as an effective cancer treatment. The efficacy of fluorinated derivatives for the treatment of several cancer modalities is well known (Cantero et al., 2006; Bronckaers et al., 2008). Floxuridine or 5-fluoro-2′-deoxyuridine has shown activity in patients with colorectal, pancreatic, breast, head, and neck cancers (Liu et al., 2008). Many studies have demonstrated that Inositol monophosphatase 1 5-chloro-2′-deoxyuridine is useful in cancer treatment (Morris, 1993). Moreover, 5-fluoro-2′-deoxyuridine and 5-chloro-2′-deoxyuridine have been useful as substrates to design new prodrugs (Johar et al.,

2005; Park et al., 2009). Biocatalysis is frequently recognized as superior to conventional chemical methods in selective modification of polyfunctional substrates owing to high catalytic efficiency, inherent selectivity, and simple downstream processing. In addition, biotransformations take place under very mild conditions and offer environmentally clean technologies (Qian et al., 2008). Transglycosylation is catalyzed by nucleoside phosphorylases. These enzymes catalyze reversible phosphorolytic cleavage of N-glycosidic bonds of nucleosides without addition of ATP, to form a free base and its respective activated pentose moiety, which is then coupled to the desired modified base to give a nucleoside analogue (only β-anomer; Bzowska et al., 2000). Halogenation is usually applied to organic structures in order to confer or enhance antitumoral activity.

, 2008). Enhanced green fluorescent protein (eGFP) was used for tagging A. brasilense strains (Wisniewski-Dyé et al., 2011). To construct egfp-containing strains, both A. brasilense strains were transformed by biparental conjugation using the Escherichia coli S17.1 harboring the broad range plasmid pMP2444 as the donor strain (Bloemberg et al., 2000). Transconjugants were isolated in Nfb with 25 μg mL−1 Gentamicin, and the stability of the plasmid was tested by streaking out single colonies

on Luria–Bertani (LB) medium for 80 successive generations (Carreño-López et al., 2000). Bacteria were grown on Agar Congo Red (ACR) plates (Rodríguez-Cáceres, 1982) for 5 days and then isolated typical colonies were chosen and each one was transferred to 125-mL flasks

containing 25 mL of LB (Difco) medium plus 5 mM MgSO4 and 3.3 mM Sirolimus solubility dmso CaCl2. These precultures were incubated at 30 °C with orbital agitation (100 r.p.m.) for 16 h until risen to 1.1–1.4 OD540 nm. Cells were harvested by centrifugation at 7500 g (Labnet Z300K) for 10 min, washed with phosphate buffer (66 mM), and resuspended to a final OD540 nm = 2. Cultures check details were diluted 1/100 in fresh Nfb-malic medium (Döbereiner & Day, 1976) modified to achieve a relation C : N = 2 using malic acid at 27.6 mM and supplemented with 13.8 mM NH4Cl or 13.8 mM KNO3 as N source. Two mL per well was transferred to sterile clear flat-bottom polystyrene 24-well plates (Costar) and incubated without agitation

for 5 days at 30 °C. All media used for Faj164 strain were supplemented with Kanamycin (25 μg mL−1; Sigma). For pMP2444-transformed Cytidine deaminase strains, Gentamicin (25 μg mL−1; Sigma) was also added. At 24-h (d1), 96-h (d3), or 120-h (d5) total growth, adhered plus planktonic cells were quantified by OD540nm measurements. Bacterial biofilm over walls of wells was mechanically removed and mixed with planktonic cells using sterile plastic sticks and agitation. This procedure efficiently removes biofilm and allows reading OD540nm using a micro plate reader (Spectra MR; Dynex Technologies). Also, viable bacteria were enumerated by dilution plating on ACR, using drop plate method (Herigstad et al., 2001). Biofilm formation was determined using crystal violet staining (O’Toole & Kolter, 1998). Briefly, each well was added with 0.5 mL of 0.5 % crystal violet. Plates were incubated for 30 min at room temperature, and then washed carefully three times with tap water. Dye attached to the wells was extracted with 2 mL of 33% acetic acid. OD590 nm in each well was determined using a micro plate reader. Data were normalized by total growth estimated by OD540 nm. Both pMP2444-transformed A. brasilense Sp245 and Faj164 strains grew for d1, d3, and d5 under static growth conditions as indicated above.

S1). In our previous work, we developed the ‘CRS cassette method’ to construct and combine markerless deletion mutations

(Hashimoto et al., 2005). The CRS cassette has a positive-selection marker (CmR) and two negative-selection markers (sacB+ and rpsL+) (Hashimoto et al., 2005; Kato & Hashimoto, 2008). First, the chromosome region to be deleted was replaced with the CRS cassette using a positive-selection marker and lambda red homologous recombination (Murphy, 1998). Next, the CRS cassette was removed using negative-selection markers and red recombination (Hashimoto et al., 2005). Two types of deletion mutants with and without the CRS cassette were constructed and transferred to recipient cells by transduction selleck chemicals with P1 phage. To avoid creating strains with synthetic growth defects and circumvent the complications associated with the use of a long CRS cassette fragment, a convenient method (ApR-415S Sm system) for introducing the new deletions was developed (Fig. 1). First, the ApR deletion units were constructed by replacing them with an ApR fragment that was shorter than the CRS cassette. After confirming that there was no synthetic growth defect, the ApR fragment was removed using ‘the 415S Sm system’ (Kato & Hashimoto, 2008), in which Cell Cycle inhibitor the chromosomal regions flanking the region to be deleted were cloned into a ts replication plasmid 415S Sm to yield ‘the deletion plasmid.’ The 415S Sm plasmid was constructed by inserting

a negative-selection marker, the wild-type rpsL allele, into the ts plasmid pHSG415S that harbored a positive-selection Casein kinase 1 marker CmR (Hashimoto-Gotoh et al., 1981). The deletion plasmid was then introduced into the

rpsL (SmR) mutant with an ApR deletion unit and the CmR transformants were selected at 42 °C. The transformants were incubated at 30 °C to obtain the SmR ApS strains. It was sometimes difficult to isolate ApS strains from the SmR strains using the ApR-415S Sm system. To make this easier, the FRT4 system was used (Fig. 2), in which a CmR fragment containing an FRT site, a recombination site for the FLP site-specific recombinase, was replaced with the deleted region to construct the CmR–FRT deletion unit. The deletion plasmid was constructed by inserting the fragment with the wild-type rpsL allele and the joined chromosomal regions that flanked the deleted regions into the plasmid pSG76A (ApR), which is an R6k derivative plasmid that lacks the pir gene required for replication (Posfai et al., 1997; Kato & Hashimoto, 2008). The deletion plasmid was introduced into the rpsL (SmR) mutant that harbored a CmR–FRT deletion unit and ApR transformants were selected. The FLP-containing plasmid was introduced into the ApR recombinant and, after adding tetracycline to the culture media to induce FLP recombinase, SmR strains were obtained (Posfai et al., 1997). Previously, a series of large-scale chromosome deletion mutants (Δ1–Δ16) were constructed.

Only one ALT per individual was measured and significant intra-individual variability with a single measure is likely. However, such misclassification is likely to be nondifferential with respect to the other factors we have considered and potentially underestimates of the associations is likely to have occured. Finally, we did not have the power to assess higher ALT elevations which might be of more clinical significance in liver disease. Notwithstanding the limitations, our study has significant strengths. Its large

selleck chemicals llc sample size allows for very accurate estimates of the relative risks for elevated ALT among ART-naïve HIV-infected individuals in comprehensive multivariate models. We were able to consider a large set of variables as determinants for elevated ALT, after adjusting for many potential confounders. Patients in the study were recruited from all the three municipalities in Dar es Salaam, increasing the external generalizability of the study. This

is a first Selleckchem AZD6244 pre-ART investigation of factors associated with ALT elevations among HIV-infected patients in a resource-limited setting, and the findings will contribute to improved patient management in such settings. In conclusion, modest elevations of ALT among ART-naïve HIV-infected patients are not uncommon in Tanzania. These elevations are more likely to occur among men, immunocompromised patients and those with components of the metabolic syndrome. These findings have important implications for long-term outcomes among HIV-infected individuals, given the known association between elevations in ALT and liver-related morbidity and mortality [6]. Longer follow-up is needed to assess the effect of elevations in ALT at baseline on morbidity and mortality in this cohort, as well as closer monitoring of ALT after initiation of ART, especially with potentially hepatotoxic therapies. We are grateful to the

patients who agreed to participate in this study. We also acknowledge the efforts all the personnel who contributed to completion of the study. HIV clinics in this study were funded in collaboration by the Government of Tanzania and the US President’s Emergency Program for AIDS Relief (PEPFAR). The first author was supported by Teicoplanin NIH-Fogarty scholar program grant no 5R24TW007988. Conflicts of interest: There is no conflict of interest. “
“The aim of the study was to assess the prevalence, predictors and patterns of genotypic resistance mutations in children after failure of World Health Organization-recommended initial nonnucleoside reverse transcriptase inhibitor (NNRTI)-based treatment regimens. We carried out a multicentre retrospective study of genotyping tests performed for all HIV-infected children at eight paediatric centres in Thailand who experienced failure of NNRTI therapy at a time when virological monitoring was not routinely available.