An autoclaved control

An autoclaved control buy SP600125 was run in parallel which consisted of 30 μM HMX added to 7-mL autoclaved WRF. Tubes were incubated anaerobically in the dark at 39 °C on a rotary shaker (150 r.p.m.); samples were taken at 0.25, 1, 2, 3, 4, and 24 h. All controls and tests were repeated in triplicate. Each strain was incubated with a concentration of 17 μM HMX, added as a liquid solution, which equaled roughly half of the dose in WRF microcosms, in low nitrogen basal (LNB) and

low carbon basal (LCB) media (Eaton et al., 2011; upon pilot testing a dose range of HMX, 17 μM was found to be the highest dose the cultures could tolerate for the 7-day incubation period). A media control consisted of 17 μM HMX in both LNB and LCB without the addition of test organism. A solvent control consisted of both types of media with 1.0 mL of overnight culture of the test organism and the addition of 0.1 mL acetonitrile. Cultures were incubated anaerobically, in the dark, at 39 °C on a rotary shaker (150 r.p.m.) for 120 h. Samples were collected at 0, 1, 4, and 5 days and processed for analysis by HPLC and LC-MS/MS as described below. Extracted samples were analyzed immediately by HPLC or frozen at −20 °C until LC-MS/MS analysis. All controls and tests were repeated in triplicate. WRF samples were collected, then frozen at −20 °C until prepared

for HPLC and LC-MS/MS analysis through solid-phase extraction using Waters Oasis HLB (3 mL/60 mg 30 μm) cartridges (Milford, MA), per the manufacturer’s instructions, Rho and modified as previously described (Eaton et al., 2013). HPLC analyses were used to determine the HMX concentration of samples and were carried out using minor modifications (Eaton et al., 2013) to Environmental Protection Agency method 8330A (U.S. Environmental Protection Agency, 2007). LC-MS/MS analyses were performed on an ABI/SCIEX (Applied Biosystems, Foster

City CA) 3200 QTRAP LC-MS/MS system using atmospheric pressure chemical ionization in the negative ion mode (Borton & Olson, 2006). A Phenomenex Ultracarb ODS (20) column (250 × 4.6 mm i.d., 5 μm particle size) was used to separate HMX and its metabolites at a flow rate of 0.75 mL min−1 over 20 min using mobile phases consisting of 0.6 mM ammonium acetate in water (A) and methanol (B) as follows: 0–5 min 90% A, decreasing linearly from 5 to 8 min to 80% A, then to 42% A from 8 to 20 min. Data were acquired using multiple reaction monitoring (MRM), using 46  355 and 147  355 (HMX + CH3COO−), 59.8  135 (methylenedinitramine), 61  118 (NDAB) as transitions. Source and gas parameters followed those in Eaton (2013). Declustering potential, entrance potential, collision entrance potential, collision energy, and collision exit potential were as follows: HMX (−15, −3.5, −24.8, −12, −4 for both transitions), methylenedinitramine (−10, −2.5, −10, −16, −58), 4-nitro-2,4-diazabutanal (NDAB; −5, −3.5, −6, −10, 0).

6% perceived the risk as high and 39% gave the risk as unknown

6% perceived the risk as high and 3.9% gave the risk as unknown. Pre-travel health advice was sought by 82% (n = 169) of those with a perceived high malaria risk at destination, by 54% (n = 54) of those with a perceived low risk, and by 41% (n = 7) of those with a perceived absent malaria risk (p = 0.001, data not shown). As shown in Table 4, the proportion of travelers carrying prophylaxis differed depending on the actual risk of malaria

at destination (p < 0.001). A company source of advice was positively associated with carrying malaria prophylaxis to high-risk (RR = 2.30, 95% CI: 1.18–4.49) and low-risk (RR = 3.12, 95% CI: 1.04–9.37) destinations (Table 2). However, FBT who received company advice were also more likely to carry malaria prophylaxis when it was not necessary to do so (ie, when traveling to no-risk destinations; RR = 3.87, 95% CI: 1.22–12.30): one in five of these travelers Selleck BGB324 were unnecessarily carrying malaria prophylaxis (Table

2). The proportion of travelers carrying an appropriate anti-malaria drug regimen was positively associated with receiving company advice among those traveling to high-risk destinations (RR = 2.10, 95% CI: 1.21–3.67), but not for those traveling to low- or no-risk destinations. Sixty-eight percent (n = 119) of travelers to a high-risk area were BMN 673 cell line carrying an appropriate anti-malaria drug regimen; for travelers to low-risk areas this was only 21% (n = 9). Advice as to which tablets to use was buy Ibrutinib provided in 68.4% by the company (occupational health physician or nurse). The company Intranet was used as a sole source by 6.6% and an additional 9.2% used multiple sources, but this always included an occupational health source of information. The remainder (9.2%) used miscellaneous sources and 6.6% did not specify the source. Most anti-malarials

were taken for prevention (75.3%), 2.5% for standby treatment, and 22% for both reasons. During the time this study was conducted, the occupational health department did not advise standby emergency treatment. Atovaquone/proguanil was by far the most commonly reported drug (44.6%), followed by mefloquine (14.3%), chloroquine (21.5%), and proguanil (14.8). Quinine (3.5%) and halofantrine (1%) were much less common. No one reported the use of doxycycline or artemether/lumefantrine. The reasons why FBT traveling to a malarious area did not carry malaria prophylaxis varied widely. There was no significant difference in carrying prophylaxis between FBT traveling to rural, urban, or beach destinations (Table 4). The majority stated that they were advised not to take tablets (39.5%). The second largest group (22.5%) judged that it was not necessary; 14% said they did not know why; for 13% the answers were very miscellaneous, and 7% had a dislike for all tablets in general. All other categories such as “I took the risk,”“prophylaxis not being deemed effective,”“forgetfulness,” and “allergy” contributed less than 6%.

The test is licensed for the near-patient detection of HIV on who

The test is licensed for the near-patient detection of HIV on whole blood, finger-prick blood and oral fluid transudate. The FDA approved the test for home use with oral fluid in the USA in July 2012 [5]. In the UK and Europe, the test is presently licensed for medical personnel use only. The manufacturer’s specificity claim is 100%

[95% confidence interval (CI) 99.7–100%] for whole blood and 99.8% (95% CI 99.6–99.9%) for oral fluid [6]. The test has been widely used in developed and resource-poor settings. From 2009 to 2010, the Department of Health-funded HIV Testing in Non-traditional Settings (HINTS) study investigated the feasibility and acceptability of routine HIV testing in general medical settings in areas of high community HIV seroprevalence in London,

UK. More than 4100 HIV tests were conducted [7]. In three of the four clinical areas studied (an emergency FG-4592 cell line department, a dermatology out-patient clinic and a primary care centre), patients would not necessarily undergo venepuncture for other indications and it was feared that blood sampling may act as a disincentive to accept an HIV test; thus, oral fluid was felt to be an appropriate specimen for HIV testing. Concerns were Talazoparib raised in each of the participating clinical areas that the use of an oral fluid POCT might have negative implications. In the emergency department, the use of a POCT with a turnaround time of 30 min did not sit well with patient pathways and strict time targets. In all clinical areas, concerns regarding the specialist training required to perform and read POCTs were cited, as was the requirement for access to specialist services 24 hours a day, in G protein-coupled receptor kinase the event of reactive tests. Pre-study patient surveys suggested that potential participants in the nonspecialist areas did not have a strong

preference for POCTs over laboratory tests. In light of the issues raised above, we resolved to develop an oral fluid-based HIV testing methodology utilizing the field collection of oral fluid specimens which were then passed on to a central laboratory for testing. Patients would be afforded the benefits of an oral fluid methodology, and participating centres need be concerned only with the safe collection of specimens in the field, obviating the need for specialist training and 24-hour referral pathways. The methodology needed to be robust, with good performance characteristics for the detection of HIV infection in low-prevalence settings, and able to handle large volume throughput. The turn-around time needed to be less than 7 days, to ensure prompt delivery of results to patients. All patients would receive their result by text message or telephone call. This paper sets out to describe our experiences of developing such a test. The development of the oral fluid HIV test falls into three phases: (1)  pre-automation oral fluid testing; In the initial phase of the HINTS study, a manual methodology was developed.

Over the last 3 years, four of them have required liver transplan

Over the last 3 years, four of them have required liver transplantation for liver failure and portal hypertension [2,3]. Examination of the explants showed a typical aspect of nodular regenerative hyperplasia related to diffuse obliterative

portal venopathy, as shown in Figure 1. Areas of hepatoportal sclerosis (HPS) were also seen in the explants. For this reason, we prefer the term ‘HIV-associated obliterative portopathy’ (HIV-OP), which better describes the syndrome of NCPH in HIV-positive patients than do the terms HPS, nodular regenerative hyperplasia (NRH), and idiopathic portal hypertension that can be found in the literature. All of these terms refer more to the consequence than to the cause of HIV-associated

NCPH [4,5]. In view of these findings, we have Sirolimus nmr PTC124 cell line screened all of our patients for coagulation abnormalities and found, in an unexpectedly high proportion of patients, a protein S (PS) deficiency [median PS level 56% of normal (IQR 46–59)] secondary to the abnormal presence of anti-PS immunoglobulin G (IgG) neutralizing antibodies [2]. We believe that the accuracy of the use of PS (activity or antigen) to diagnose early HIV-OP should be assessed. As our data suggest a prothrombotic state, use of anticoagulants is also an important issue that should be addressed in a clinical study, as oral anticoagulants Phosphoglycerate kinase are effective in preventing thrombosis in congenital PS deficiencies.

“A cold shock domain (CSD)-containing protein, CspD, of molecular mass ∼7.28 kDa in a psychrotolerant Antarctic Janthinobacterium sp. Ant5-2 (ATCC BAA-2154) exhibited constitutive expression at 37, 22, 15, 4 and −1 °C. The cspD gene encoding the CspD protein of Ant5-2 was cloned, sequenced and analyzed. The deduced protein sequence was highly similar to the conserved domains of the cold shock proteins (Csps) from bacteria belonging to the class Betaproteobacteria. Its expression was both time- and growth phase-dependent and increased when exposed to 37 °C and UV radiation (UVC, dose: 1.8 and 2.8 mJ cm−2). The results from the electrophoretic mobility shift and subcellular localization study confirmed its single-stranded DNA-binding property. In silico analysis of the deduced tertiary structure of CspD from Ant5-2 showed a highly stable domain-swapped dimer, forming two similar monomeric Csp folds. This study established an overall framework of the structure, function and phylogenetic analysis of CspD from an Antarctic Janthinobacterium sp. Ant5-2, which may facilitate and stimulate the study of CSD fold proteins in the class Betaproteobacteria. Microorganisms isolated from Antarctica are suitable candidates to study physiological and genetic mechanisms for the adaptation to cold and subzero temperatures.

Unless otherwise stated, experiments were performed using wild-ty

Unless otherwise stated, experiments were performed using wild-type C57BL/6J (Jackson Laboratories) or ICR (Harlan Laboratories) animals mated in house to generate timed pregnancies. Experiments to test Cre expression used Ai3 ROSA26 CAG-lox-stop-lox-eYFP [Jackson Laboratories stock #7903 (Madisen et al.,

2010)] or the R26R lacZ reporter line [Jackson Laboratories stock #3474 (Soriano, 1999)]. Experiments to test tTA expression used the tetO-nls-GFP-lacZ reporter line (Mayford et al., 1996). Transgenic offspring for these experiments were generated by mating Ai3, R26R or tetO-nls-GFP-lacZ males with ICR females. The viral injections 17-AAG cell line described below were performed blind to genotype, and transgenic status determined by tail biopsy at either the time of weaning or harvest. All procedures were reviewed and approved by the Baylor College of Medicine Institutional Animal Care and Use Committee in accordance with the guidelines of the U.S. National Institutes of Health. Within 6 h of birth, neonates were collected from the cage and cryoanesthised at 0 °C for 3 min before injection. Following cessation of movement, a solution of recombinant AAV diluted in sterile phosphate-buffered saline containing 0.05% trypan blue was injected bilaterally into the ventricles using a 10 μL Hamilton syringe (Hamilton, 7653-01) with a 32 gauge needle (Hamilton, 7803-04, RN 6PK PT4). The

injection site was located two-fifths of the distance along a line defined between

each eye and the lambda intersection of the skull (Fig. 1). Z-VAD-FMK datasheet The needle was held perpendicular to the skull surface during insertion to a depth of approximately 3 mm. Once the needle was in place, 2 μL of viral solution was manually injected into each lateral ventricle [1 μL for experiments comparing postnatal day (P)0 and adult injection]. After both injections were complete, pups were placed on a warming pad until they regained normal color and resumed movement. All injected animals were then transferred to an ICR foster mother for care. The ICR foster mothers had delivered within 4 days before the day Thalidomide on which the pups were injected. Depending on the number of injected pups needing care, most or all of the pups born to the ICR foster mothers were removed to ensure success of the injected animals. For delayed injection experiments, P1 (24–30-h-old), P2 (48–54-h-old) or P3 (72–78-h-old) neonates were injected as above. Adult mice (2–4 months) were anesthetised with 1.5% isoflurane, placed in a stereotaxic apparatus, and prepared for viral injection by a midline scalp incision followed by the opening of a small burr hole in the skull over the desired injection site at 1.5 mm caudal to the bregma, 0.5 mm lateral to the midline, and 1.3 mm deep to the dura mater. A volume (1 μL) of AAV diluted in phosphate-buffered saline containing 0.

In vitro studies have shown

In vitro studies have shown Vemurafenib increased expression of HPV E1 and L1 viral genes in the presence of HIV transactivator

of transcription (tat) proteins [17]. Our analysis using an older set of high-risk HPV types suggested that higher VL may be associated with HPV detection and hinted at a role of HIV VL in HPV acquisition. However, such an association was not suggested using the latest set of high-risk HPV types. Hence, our research demonstrates the importance of considering the HPV types used when reviewing the literature. It has been postulated that immune reconstitution associated with HAART may lead to clearance of HPV, as has been the case with other viral or nonviral opportunistic infections,

and this is consistent with the results of our analyses, where higher CD4 cell count was associated with a higher probability of HPV clearance. There are a couple of limitations to our analyses. There was a trend for earlier discontinuation in subjects starting with HPV infection, suggesting possible informative censoring, and the small sample size did not allow the use of models that adjust for covariates such as age, cigarette smoking, HPV type and sexual activity. There are several advantages of the statistical methods that we used. The multi-state modelling approach accommodates multiple and recurrent events using intermittent SB431542 solubility dmso data. The hazard rates are estimated simultaneously in the model, eliminating the need to subset the data to estimate the HPV detection rate among 4��8C HPV-negative subjects

and separately to estimate clearance among the HPV-positive subjects. Also, while other HPV studies have used the midpoint between visit times as the event time (e.g. the time of HPV detection or clearance) or the time of the visit, the methods we used are appropriate when the exact event times are unknown. The approach utilized the incomplete data efficiently and provided a more comprehensive description of the HPV detection and clearance process. We thank the clinicians, study coordinators and study subjects at A5029 sites for their participation and the A5029 study team, headed by Ken Fife, for sharing the data. We also thank Stephen Lagakos, Janet Andersen and Michael Hughes for their thoughtful comments on the analysis. The authors are supported by the AIDS Clinical Trials Group and K24 Mid-Career Research Mentoring Award funded by the National Institute of Allergy and Infectious Diseases (Grants 1U01AI068636-01, 1U01AI068634-01 and K24AI066884).

5, range 55–10) Seven nursing staff reported spending less than

5, range 5.5–10). Seven nursing staff reported spending less than 10 min to visit a patient and conduct the INR test. Nurses most commonly reported spending a further 2–5 min entering the result into the MedePOC system (four nurses), with a three nurses spending between 5 and 10 min. Four nurses indicated that GPs generally responded within 24 h of receiving the INR test; two nurses indicated that responses typically came within 24–48 h. The patients who responded to

the evaluation questionnaire were all satisfied with their ACF’s involvement in the study (median score 9, range 5.5–9.5). Most patients indicated they would prefer a finger-prick blood test with the portable INR monitor to the usual venous blood sampling selleck screening library taken by the pathology service (median score 8.5, range 0.5–9.5), and that they would prefer their INR to be

monitored at the nursing home rather than through pathology testing (median score 9.3, range 0.5–9.5). All patients felt that their warfarin was better controlled during the study (median score 9, range 6.5–9.5). This proof-of-concept study was designed to test the utility of ACF-based INR testing with electronic communication to GPs. Weekly POC INR monitoring conducted in ACFs with electronic communication of results to GPs and warfarin doses back to the ACFs resulted in non-significant improvements in INR control. An improvement in warfarin control was shown for the majority of patients, and POC monitoring was well received by nursing staff, suggesting that further research is warranted to investigate whether this strategy can improve INR control in this population. Several factors may limit the generalisability of this study. We included a selected group of participants

who may not be representative of the broader population of older people taking warfarin. It is possible that they were more motivated and more adherent Tacrolimus (FK506) with their therapy than other patients.[23] Indeed, the intervention may be more successful in people with poorer prior INR control, as the participants had a reasonable level of control prior to the intervention. Other potential limitations include the small sample size, non-randomised design and relatively short duration of the intervention. Certainly, further investigation of the system in patients taking warfarin is warranted, including an evaluation of the costs involved and effectiveness of the approach in a larger cohort. While we have reported on the implementation of the system in a small number of ACFs, a number of challenges would need to be addressed if it was to become more widely used, including ongoing education of nursing staff, quality assurance of the testing procedure and auditing of the communication process to ensure appropriate response to INR test results.

This indicates that a classifier trained only on pictures of sepa

This indicates that a classifier trained only on pictures of separately presented faces and places may not be the most optimal way of decoding object-based visual attention. Concluding, we have shown that real-time fMRI allows for online prediction of attention to objects belonging to different object categories. Prediction is based on distributed patterns of activity in multiple brain regions. The outlined methodology not only allows us to probe object-based attention in an online setting EPZ015666 mouse but also illustrates the potential to develop BCIs that are driven

by modulations of high-level cognitive states. The authors gratefully acknowledge the support of the BrainGain Smart Mix Programme of the Netherlands Ministry of Economic Affairs and the Netherlands Ministry of Education, Culture and Science. The first

author was supported by a UTS grant from the University of Twente. We thank Paul Gaalman for his technical support during the experimental setup and development of the real-time fMRI pipeline. We are very grateful to the editors and the anonymous reviewers for their encouraging and constructive comments on our manuscript. Abbreviations aMTG anterior medial temporal gyrus BCI brain–computer interface BOLD blood oxygen level-dependent FFA fusiform face area fMRI functional magnetic resonance imaging GLM general linear model MoCo motion-corrected MVA-C cluster-wise anti-PD-1 antibody multivariate analysis MVA-G GLM-restricted multivariate analysis MVA-T mean timeseries multivariate analysis MVA-W whole-brain multivariate analysis MVPA multivoxel pattern analysis OFA occipital face area PACE prospective acquisition correction TR repetition time

Fig. S1. A basis set of 15 face-place pairs used in decoding phase. Each pair was used twice in each condition, once with the face picture set as target and the other time with the place picture set as target. Note: Copyrighted pictures used in the original experiment have been replaced in the above graphic by their non-copyrighted look-alikes. Fig. S2. Graph-based visual saliency algorithm was used to select the face-place pairs. Saliency of the 50/50 hybrid and each of its constituents were Carbohydrate observed and only those pairs were selected for which the 50/50 hybrid had an equal number of salient points for the face and place picture. Fig. S3. Stimulus timeline. (A) Example of an attend-face trial in non-feedback condition. (B) Example of an attend-place trial in feedback condition. After cues have been presented, the face-place hybrid image was updated every TR depending on classification result of the preceding TR. Fig. S4. List of all brain regions from which voxels were selected by the MVA-W classifier for training. Only regions that were activated across three or more subjects were used for further analyses. Fig. S5.

3 mM 4-DPS and 200 μL 1 M DTT, respectively, and was performed fo

3 mM 4-DPS and 200 μL 1 M DTT, respectively, and was performed for each sample. The fluorescence of the cells was detected in 96-well plates (FluoroNunc, Nunc) on a fluorescence

photometer (Spectramax GeminiXS, Molecular Devices, Sunnyvale, CA), where it was possible to measure the excitation of two wavelengths, namely 395 and 465 nm, corresponding to the oxidized and the reduced form of the protein. Each sample was measured four times. The fraction of oxidized roGFP (Ox) was calculated according to the following equation: (1) It is comprised of the fluorescence of fully oxidized (Fox) or reduced (Fred) cells and the untreated sample (F), respectively. Thiazovivin The genes of roGFP1, roGFP1_iE and roGFP1_iL were expressed in either cytosol or ER of the P. pastoris strain X-33. The ability of the three constructs to monitor redox changes in different compartments of living yeast cells was examined through comparison of the SD of the redox ratios and the range selleck chemicals llc between the total reduction and the total oxidation of the respective roGFP (Fig. 1a–d). roGFP1 sensors are ratiometric by excitation, which means that they exhibit two excitation peaks at about 395 and 465 nm, corresponding to the neutral and the anionic chromophore forms, respectively, with a single emission peak at 510 nm (Lohman & Remington, 2008). This ratiometric behavior is

advantageous, because the redox determination is independent of the concentration of the roGFP. Fluorescence measurements were taken after the treatment of all transformants with DTT and 4-DPS as described above and compared with the fluorescence of untreated cells of the same transformants. Therefore, an external calibration was not necessary. As can be seen in Fig. 1a, the observed variability in the cytosol is low for roGFP1, but much higher for the ER-optimized constructs. In the ER, roGFP1 is fully oxidized (as observed previously by Schwarzer et al., 2007; Merksamer et al., 2008), in contrast to roGFP1_iE and roGFP_iL, which are only 45–50% oxidized (Fig. 1b). According to the wider range between

totally oxidized and totally reduced states, roGFP1_iE was chosen for the determination of the redox state in the ER (Fig. 1d). The localization of the two chosen constructs for cytosol and ER was analyzed by taking images using a confocal O-methylated flavonoid laser microscope. An ER pattern was present for roGFP1_iE targeted into the ER, whereas cytosolic roGFP1 was distributed all over the cell, except for the vacuole, without showing a distinct pattern (data not shown). The reduction potential was determined using Eqn. (3) derived from the Nernst equation, and the midpoint potentials E°′roGFP for roGFP1 (−287 mV), roGFP1_iE (−236 mV) and roGFP1_iL (−229 mV), respectively (Lohman & Remington, 2008): (3) According to this calculation, the cytosol of P. pastoris X-33 has a reduction potential of −295 mV.

A UNG enzyme step (50 °C for 2 min) ensured hydrolysis of all sin

A UNG enzyme step (50 °C for 2 min) ensured hydrolysis of all single-stranded and double-stranded contaminating PCR products. Cycle threshold (CT) values >40 cycles were considered negative. The sensitivities of the IS2404/IPC and the IS2606/KR multiplex assays achieved in this setting were compared with the values described by Fyfe et al. (2007) by performing real-time PCR on serial dilutions of purified M. ulcerans DNA. Like Fyfe et al. (2007), our assays reliably detected two copies of IS2404, nine copies of IS2606,

and 1.5 to three copies of KR. We studied the effects of postponing a run of a prepared reaction plate on assay PARP inhibitor sensitivities in a similar way by keeping prepared plates at 4–8 °C for a period of >12 h before real-time PCR analysis was carried out, simulating the effects of a possible power cut before analysis could be started. This delay in analysis did not alter the sensitivities of the assays in any way. Pooled organs of 62 small mammals (36 Praomys spp., 10 Mastomys spp., five Lemniscomys spp., three Lophuromys spp., four Crocidura spp. Selleckchem GSK126 and four Mus spp.) caught in houses and around water bodies of a BU-endemic village (Ananekrom, in the Ashanti Region of Ghana; Fig. 1) as described before (Durnez et al.,

2008) were analyzed after DNA extraction using the modified Boom method (Boom et al., 1990; Durnez et al., 2009). Although none of the PCR reactions were inhibited, IS2404 was not detected in any of the specimens. A total of 148 environmental samples (13 water samples, 45 detritus samples,

45 trunk biofilm, and 45 plant biofilm samples) collected from water bodies near five BU endemic villages (n=117) and two BU nonendemic villages (n=31) (Fig. 1) were also analyzed. Although the DNA extraction procedure included a purification step using diatomaceous earth, reactions in 50 of the 148 environmental specimens were inhibited as they had CT values of the IPC three cycles higher than the nontemplate controls. These inhibited samples were successfully reanalyzed with a newly developed environmental master mix adapted for real-time PCR-based Glycogen branching enzyme detection in the presence of high levels of common environmental inhibitors (Applied Biosystems, TaqMan® Environmental Master Mix 2.0, ref. 4396838). Three samples (2.0%) were positive for IS2404, with CT values of 36.31, 38.45, and 37.95, respectively (Table 1). Of the three positive samples, only the water sample from Nshyieso also tested positive for IS2606 and KR, with a ΔCT (IS2606-IS2404) value of 1.96 (Table 1), suggesting that M. ulcerans DNA was detected and not DNA from other IS2404-containing mycobacteria that are known to have higher ΔCT values (Fyfe et al., 2007). The CT (IS2404) values of the other two IS2404-positive samples were higher than the sample that did contain IS2606 and KR, suggesting that the failure to detect KR and IS2606 was caused by a low DNA concentration, which is consistent with known differences in copy number per cell.