Data regarding the 131I content in these 28 women and relevant information released by the citizens group on April 21 and May 18 were obtained from their website (‘Radioactivity in breast milk’, cited September 15, 2011; available from URL: http://bonyuutyousa.net/). Air pollution with radioactive materials occurred over a geographically wide area within 300 to 400 km of the FNP in the morning of March 15, 2011 (Fig. 2). Although the air radiation

dose rate was <0.07 µGy/h before the FNP accident in the areas shown in Figure 1, it increased sharply to 19 µGy/h in Fukushima city on March 15, then decreased to 1.6 µGy/h by the end of May. In Tokyo, located 230 km south of the FNP, the highest radiation dose rate of 0.81 µGy/h on March 15 decreased to <0.07 µGy/h by mid-April. The amount http://www.selleckchem.com/products/Y-27632.html of 131I radioactivity in fallout per day reached a peak level of 93 000 MBq/km2 in Hitachinaka

city, located 130 km south of the FNP, on March 20, while it reached a peak level of 38 000 MBq/km2 in Tokyo on March 22 (Fig. 3). Consequently, vegetables such as spinach, cows milk and chicken eggs were also contaminated with 131I (Fig. 4). The highest content of 131I was 24 000 Bq/kg, found in spinach on March 18 in Kitaibaraki city, located 75 km south of the FNP. The 131I content in spinach decreased over time; for example, a level of 3500 Bq/kg was recorded in Utsunomiya city on March 19, decreasing to 480 Bq/kg on April 13, 120 Bq/kg on April 20, 12 Bq/kg on April 26, and became undetectable on May 3 (Fig. 4). Among the three foods, www.selleckchem.com/products/CAL-101.html the 131I content was lowest in chicken eggs. It rained on March 20 and 21 in these areas, and the rain accelerated the pollution of water with 131I (Fig. 5). In Tokyo, 131I radioactivity in tap water from the Kanamachi water

purification plant reached a peak level of 210 Bq/kg on March 22. The content of 131I in the tap water decreased and became undetectable in many cities by mid-April (Fig. 5). Seven of 23 women (30.4%) who were tested in April secreted a detectable level of 131I in their breast milk (Table 1). The concentrations ranged from 2.2 to 8.0 Bq/kg and appeared to be higher than those in tap water Nabilone available for these seven women at the same time points. As expected from the data on 131I radioactivity in the fallout, vegetables and water (Figs 3 to 5), the radioactivity of 131I in the breast milk became undetectable by May 15 in these seven women (Table 1). None of the remaining 96 women tested in May exhibited a detectable amount of 131I in their breast milk samples with detection limits of 1.6 ± 0.3 Bq/kg (data not shown). The present study demonstrated that environmental pollution with 131I causes the contamination of breast milk with 131I.

6-fold reduction in susceptibility [1–5]. A median 1.8-fold reduction in susceptibility BI 6727 in vivo to ATC has been observed in a study of HIV-1

isolates containing five TAMs in the 41, 215 pathway and a median 1.3-fold reduction in susceptibility in isolates containing five TAMs in the 67, 70, 219 pathway [3]. ATC has shown promising antiviral activity when given as monotherapy over 10 days in treatment-naïve HIV-1-infected patients [6]. In a double-blind, randomized Phase II study of 63 patients, reductions in viral load were observed in patients receiving one of four different total daily doses of ATC (given as six dosing regimens), with all dose groups having a statistically significant decrease in plasma HIV-1 RNA levels from baseline relative to placebo after 7 days of treatment. In addition, ATC did not select for any particular mutation during the 10-day treatment period. The aim of this study was to evaluate and compare the efficacy and safety of two doses of ATC in HIV-1-infected patients who were treatment-experienced and harbouring the M184V mutation, with or without additional TAMs. Patients enrolling in this study were failing their current 3TC- or FTC-containing regimen, with limited remaining

treatment options. The first part of the study (to day 21) evaluated the antiviral effect of two doses of ATC by replacement this website of the 3TC/FTC in the patients’ existing treatment regimen with one of two doses of ATC or with (continued) 3TC. The 3-week duration of this period of treatment with ATC allowed assessment of the activity of ATC in

the background of a failing treatment regimen while limiting the potential for the development of resistance mutations. At day 21, the background antiretroviral therapy (ART) could be optimized according to the patient’s genotype at screening and treatment with ATC or 3TC continued to week 24. Reported here are the primary endpoints of the study, the efficacy and safety results at day 21. SB-3CT This was a Phase IIb randomized, double-blind, dose-ranging, multicentre study conducted in Argentina and Australia. The study was conducted according to International Conference on Harmonisation (ICH) Good Clinical Practice Guidelines and was approved by the appropriate local Ethics Committees. All patients gave written informed consent before participating in the study. Eligible patients had a documented laboratory diagnosis of HIV-1 infection [positive enzyme-linked immunosorbent assay (ELISA) HIV-1 antibody test confirmed by western blot, p24 assay, HIV-1 RNA or culture] with plasma HIV-1 RNA levels≥2000 copies/mL (using the Ultrasensitive COBAS Amplicor® HIV-1 Monitor™ version 1.5, Roche Molecular Systems Inc., Branchburg, NJ, USA) and presence of the M184V mutation in the HIV-1 reverse transcriptase at screening by genotype assay.

, 1999; Degrassi et al., 2002). The results suggest that dipeptide-like compounds such as diketopiperazines are ubiquitous in a natural environment as antimicrobial agents or cell–cell communication molecules (Holden et al., 2000). A blast search revealed that the homologs of Bcr, NorE, YdeE and YeeO prevail in Enterobacteriaceae (data not shown). It was reported that cyclo (Ala-Val) was found in cell-free supernatants from Proteus,

Citrobacter and Enterobacter (Holden et al., 1999). Interestingly, these genera possess close homologs of dipeptide transporters. Taken together, dipeptide transporters and their homologs could be involved in the transport of diketopiperazines in these bacteria. Although it is too early to speculate on High Content Screening the natural functions, these multidrug-efflux transporters may transport dipeptide-like molecules to extracellular space to maintain homeostasis, or as signals for cell–cell communications. It was recently reported that the natural function of NorE was to export signals for cell–cell communication (Yang et al., 2006). In this study, two functionally uncharacterized genes, ydeE and yeeO, were identified as those conferring dipeptide

resistance. Although the minimum inhibitory selleck compound library concentration (MIC) of various antimicrobial agents and chemical compounds were examined in E. coli cells overexpressing ydeE (ydeF), no alteration of MIC was observed (Nishino & Yamaguchi, 2001). In Erwinia chrysanthemi, it is reported that yeeO is a member of the predicted regulon of KdgR, the transcriptional regulator of pectin catabolism O-methylated flavonoid (Rodionov et al., 2004). However, no other information about its function is obtained. In contrast, the effects of ydeE or yeeO overexpression on the reduction of intracellular Ala-Gln and also on the production of Ala-Gln and Ala-BCAA were remarkable (Figs 3 and 4). Bcr and YdeE are classified into the major facilitator superfamily. As shown in Table 2, E. coli cells overexpressing Bcr were resistant to bicyclomycin, tetracycline, fosfomycin, kanamycin and l-cysteine.

NorE and YeeO are classified into the multidrug and toxic compound extrusion family. NorE was identified as a quinolone resistance protein at first (Morita et al., 1998). Also, E. coli cells overexpressing NorE were resistant to chloramphenicol, doxorubicin, fosfomycin, trimethoprim, etc. (Nishino & Yamaguchi, 2001). Although the structure of known substrates is rather dissimilar to Ala-Gln or Ala-BCAA, significant effects on dipeptides production suggest that Bcr, NorE, YdeE and YeeO are involved in the transport of dipeptides. Substrate specificities of these proteins remain to be elucidated. The spectrums of dipeptide to which dipeptide transporters conferred resistance were wide and also differed correspondingly (Table S1).

, 2007). The RegSR system has long been known to activate the transcription of the nifA gene that encodes the key regulator for nitrogen-fixation

genes in B. japonicum (Bauer et al., 1998). Among the novel RegR target genes, we identified a putative operon (blr1515–blr1516) that encodes a predicted multidrug efflux system. Here, we report the characterization of a mutant lacking see more this predicted transport system, now designated BdeAB, and demonstrate that it confers antibiotic resistance and is required for an efficient symbiosis specifically with soybean. Bradyrhizobium japonicum strains were routinely cultivated in a peptone–salts–yeast extract (PSY) medium supplemented with 1 g l−1l-arabinose as described elsewhere (Regensburger & Hennecke, 1983; Mesa et al., 2008). Alternatively, we used a modified Vincent’s minimal medium (Vincent, 1970; Akt inhibitor Becker et al., 2004) that was supplemented with 3 g l−1l-arabinose, 10 mM MOPS (final pH of medium adjusted to 6.8 with 2 M NH3), and trace elements as described elsewhere (Bishop et al., 1976). When appropriate, antibiotics were used at the following concentrations (μg ml−1): spectinomycin, 100; streptomycin, 50; tetracycline, 50 (solid media) or 25 (liquid media); and cycloheximide, 100. Bradyrhizobium japonicum strain 110spc4 was used as the wild type (Regensburger & Hennecke, 1983). Mutant

derivatives relevant for this work were strain 2426 (ΔregR∷Ω; Bauer et al., 1998); strain 9589 (ΔbdeAB∷Ω; see below); and strain 9589-38 (strain 9589 complemented with chromosomally inserted wild-type bdeAB genes; see below). Escherichia coli strains were grown in Luria–Bertani medium (Miller, 1972) containing the following concentrations of antibiotics for plasmid selection (μg ml−1): ampicillin, 200; streptomycin, 50; and tetracycline, 10. Strain DH5α (Bethesda Research Laboratories, Gaithersburg, MD) was the host for cloning, and S17-1 (Simon

et al., 1983) for the conjugation of plasmids into B. japonicum. Sterilization of seeds of soybean [Glycine max (L.) Merr. cv. Williams], cowpea (Vigna unguiculata), siratro (Macroptilium atropurpureum), and mungbean (Vigna radiata), plant growth conditions, and measurement of nitrogenase activity were performed as described previously (Göttfert et al., 1990; Gourion Evodiamine et al., 2009; Koch et al., 2010). At least 107 cells of B. japonicum were added as inoculum. For bacteroid isolation, all nodules from individual soybean plants infected by either the wild type or the ΔbdeAB strain 9589 were collected and weighed. Nodule material was then crushed in PSY medium, and serial dilutions of the bacteroid suspension were spotted in four parallels on PSY agar plates containing spectinomycin and cycloheximide. After a 1-week incubation at 30 °C, the number of CFU per milligram of nodule wet weight was determined.

Although C. pneumoniae-specific antibody responses have been characterized by immunoblotting, only few major surface proteins (MOMP, Omp2, and CrpA; Iijima et al., 1994; Klein et al., JAK inhibitor 2003; Mygind et al., 1998) and some Inc proteins (Cpj0146, Cpj0147, and Cpj0308) have been detected (Hongliang et al., 2010). However, these antigens have yielded variable results with respect to the consistency and accuracy of C. pneumoniae identification. Taken together, very little information is available regarding specific detection of C. pneumoniae. We determined the sequence of the whole genome of C. pneumoniae J138 isolated

in Japan (Shirai et al., 2000) and found that this strain features putative protein coding from its 1069 open reading frames (ORFs). A comprehensive bioinformatics approach was applied for annotation taxonomy, and about half of the predicted genes were found to encode proteins without any known functions. To identify novel specific antigens from C. pneumoniae, we screened 455 genes without any known functions. A fusion protein expression library of C. pneumoniae was constructed in Saccharomyces cerevisiae. Protein extracts of the recombinant yeast cells expressing the green fluorescent protein (GFP)-tagged C. pneumoniae proteins were subjected to Western blot analysis using serum samples from C. pneumoniae-infected patients as the primary

antibodies. This study sought to identify specific and highly immunodominant antigens, which are required for the development of new serodiagnostic assays, and hopefully, vaccines, in the future. Thirteen serum samples were collected from eight patients HSP90 (age: range, 4–11 years; see more Table 1), who had been clinically diagnosed with primary acute C. pneumoniae infection. The levels of C. pneumoniae-specific immunoglobulin (Ig) IgA, IgG, and IgM in these patients were evaluated using two different

ELISA kits: (1) HITAZYME C. pneumoniae kits for IgA, IgG, and IgM that utilize the soluble elementary body (EB)-outer membrane complex, without the lipopolysaccharide, as the antigen (Hitachi Chemical, Japan) and (2) C. pneumoniae-ELISA plus Medac kits for IgA and IgG and C. pneumoniae-sELISA Medac kit for IgM, which utilize the purified cell wall membrane proteins as the antigen (Medac Diagnostika, Germany). Eight serum samples from 0-year-old healthy children were used as negative controls. Chlamydophila pneumoniae genomic DNA was obtained from the EBs of the C. pneumoniae J138-infected HEp-2 cells (Miura et al., 2001). We used a gene expression system controlled by a Tet-off promoter in S. cerevisiae. The ORFs of 455 genes from C. pneumoniae J138, including genes of unknown function (Supporting Information, Table S1), were cloned into a pMT830 vector, which was constructed as previously described (Tabuchi et al., 2009). This vector system allows a protein of interest to be expressed with GFP fused to the C-terminus.

The Mma Bana study from Botswana randomly allocated 560 women at 26–34 weeks’ gestation, with CD4 cell counts >200 cells/μL to receive either lopinavir/ritonavir plus zidovudine/lamivudine (PI group) or abacavir/zidovudine/lamivudine (NRTI group). The PTD rates were significantly higher in the PI group (21.4% vs. 11.8%; P = 0.003) [101]. A second study, the Kesho Bora Study randomly allocated 824 women at 28–36 weeks’ gestation, again with CD4 cell counts >200 cells/μL to receive lopinavir/ritonavir and zidovudine/lamivudine or zidovudine monotherapy twice

daily plus a single dose of nevirapine at the onset of labour. There was no difference in the PTD rate between the two groups (13% with PI vs. 11% with zidovudine monotherapy/single-dose nevirapine) [102]. The randomized studies above are two of few studies that selleck compound have been able to look at individual PIs. One additional analysis from the APR of 955 live births exposed to lopinavir/ritonavir reported a PTD rate of 13.4% [103]. A retrospective study from the UK reported a PTD rate of 10% in 100 women taking ritonavir-boosted atazanavir in pregnancy, of whom 67% had conceived on their regimen [79]. The data regarding HAART, individual components of HAART and PTD remain conflicting. Some studies suggest that PIs, in particular ritonavir-boosted PIs, are associated with an increased

risk of PTD but this is not confirmed by others. There is a need for a randomized study of sufficient power to explore these issues further and the Promoting Maternal and Infant Survival Everywhere (PROMISE) study (NCT01061151), with 6000 women either randomly allocated to a PI-based combination selleck screening library regimen or zidovudine monotherapy will hopefully provide some answers to these important questions. 5.2.4 No routine dose alterations are recommended for ARVs during pregnancy if used at adult licensed doses with the

exception of darunavir, which should be dosed twice daily. Grading: 1C Consider third-trimester pheromone TDM particularly if combining tenofovir and atazanavir. Grading: 1C If dosing off licence, consider switching to standard dosing throughout pregnancy or regular TDM. Grading: 1C Physiological changes that occur even during the first trimester of pregnancy may affect the kinetics of drug absorption, distribution, metabolism and elimination, thereby affecting the drug dosing. Gastrointestinal transit time becomes prolonged; body water and fat increase throughout gestation and there are accompanying increases in cardiac output, ventilation, and liver and renal blood flow; plasma protein concentrations decrease, notably albumin and α1 acid glycoprotein; renal sodium reabsorption increases; and changes occur in the metabolic enzyme pathway in the liver, including changes in cytochrome P450. Caution should be exercised if women fall pregnant on unlicensed doses and consideration given to performing TDM to assess trough levels, or reverting to licensed dosing, often twice per day, during pregnancy.

, 1997). This polyP accumulation is due to the inhibition of polyP degradation by ppGpp rather than the loss of PhoU function (Kuroda & Ohtake, selleck products 2000; Kuroda, 2006). To determine whether YjbB reduces the levels of polyP under conditions of amino acid starvation, we introduced pMWyjbB into the wild-type strain and then subjected the transformant to amino acid starvation. The levels of polyP in the transformant were lower than those of the strain carrying a control vector plasmid (Fig. 2a). Escherichia coli also accumulates polyP when its growth is blocked by antibiotics that inhibit nucleic acid synthesis

(Kuroda & Ohtake, 2000; Kuroda, 2006). When treated with rifampicin, the levels of polyP in the transformant were also lower than those in the strain carrying a control vector plasmid (Fig. 2b). These results also supported the hypothesis that the reduction of polyP was not due to the suppression of the expression of Pho regulon genes including pstSCAB. As noted above, the N-terminal half of YjbB shows homology with Na+/Pi selleck chemicals cotransporters, indicating the possible involvement of YjbB in the Pi flux. Escherichia coli possesses four Pi transporters (PitA, PitB, PhnCDE, and PstSCAB). Here, we constructed a mutant strain, MT2006, which lacks all four Pi transporters (Table 1). This mutant lost the ability to grow on a medium containing

Pi as the sole source of phosphorus (Pi medium) (Fig. 3a). To test whether YjbB is involved in Pi import, we introduced pMWyjbB into MT2006. However, this transformant still failed to grow on the Pi medium (Fig. 3a). Escherichia coli can utilize glycerol-3-phosphate as the sole source of Pi (Hayashi et al., 1964; Schweizer et al., 1982). The transformant could grow on a medium containing glycerol-3-phosphate as the sole source of phosphorus (GP medium) (Fig. 3b), indicating that YjbB has no or little Pi-uptake activity. On the other

hand, the transformant released approximately 1 mM Pi into the supernatant Methane monooxygenase when it grew for 8 h on the GP medium. To exclude the possibility that Pi was due to the degradation of glycerol-3-phosphate by an elevated alkaline phosphatase activity in the phoU mutant, we constructed MT2013 (phoA, yjbB, pitA, pitB, phnC, pstSCAB-phoU). Similar to MT2006, MT2013 and its transformant harboring pMWyjbB lost the ability to grow on Pi medium, but could grow on GP medium (Fig. 3). MT2013 carrying pMWyjbB still released a large amount of Pi into the GP medium, while MT2013 carrying a control vector plasmid only released a small amount of Pi during the lag phase (Fig. 4a and b). Escherichia coli can take up glycerol-3-phosphate via glycerol-3-phosphate transport systems (Ugp and GlpT) (Hayashi et al., 1964; Schweizer et al., 1982). The GlpT transport system can function in the exchange mode, so that glycerol-3-phosphate is taken up in exchange with internal Pi, while the Ugp system does not release Pi.

The CCR is both

a registry at every VA facility to support local care delivery and a national clinical database. The CCR database aggregates data from all facilities to the unique patient level. It compiles very detailed data on HIV-infected patients’ demographics, diagnoses, laboratory tests, and clinic and drug utilization. For the current analysis, only patients who entered the registry in the HAART era (1996–2004) were included. We used diagnostic codes and HCV antibody tests to identify patients with HCV coinfection. We included the following International Classification of Diseases (ICD-9) codes when they appeared as one of the listed discharge diagnoses: 070.41, ‘acute hepatitis C with hepatic coma’; 070.44, ‘chronic Compound C nmr hepatitis C with hepatic coma’; 070.51, ‘acute hepatitis C without mention of hepatic coma’; 070.54, ‘chronic hepatitis

C without mention of hepatic Z-VAD-FMK mouse coma’; V02.62, ‘hepatitis C carrier’. A validation study previously showed that the presence of an HCV code was 94% predictive of a positive HCV laboratory test result, while the absence of a code was 90% predictive of the absence of a positive test result. Of all patients with HIV infection in the VHA CCR, 96% were tested for HCV [31]. Lipid profiles were extracted from each patient’s records, including TC and TG levels. For patients with more than one measurement of the lipid profile during the study period, the measurement with the highest level of TC and TG was used, regardless Evodiamine of lipid-lowering therapy history. These laboratory measures were used to classify patients as having hypercholesterolaemia and/or hypertriglyceridaemia. The proportion of patients with other known cardiovascular risk factors, including hypertension, type 2 diabetes mellitus and smoking status, was calculated in HIV/HCV-coinfected and HIV-monoinfected patients. Patients’ records

were reviewed for the presence of the following ICD-9 codes when they appeared as one of the listed discharge diagnoses: 401, ‘essential hypertension’; 250.0, ‘diabetes mellitus without mention of complication’ (except when the fifth digit was 1 or 3, indicating ‘type 1 diabetes mellitus’); 250.1 to 250.9, ‘diabetes mellitus with complications’; 305.1, ‘tobacco use disorder’; V15.82, ‘history of tobacco use’. Data on the use of antiretroviral and lipid-lowering medications were also extracted. We calculated the duration of use of medications by estimating the number of days covered by each prescription. We report on two outcomes: incident acute myocardial infarction (AMI) and incident cerebrovascular disease (CVD; transient ischaemic attacks or strokes). We included the following ICD-9 codes when they appeared as one of the listed discharge diagnoses: 410, ‘AMI,’ except with a fifth digit of 2 (indicating a subsequent instead of initial episode of care); 433, ‘occlusion and stenosis of precerebral arteries’; 434, ‘occlusion of cerebral arteries’; 436, ‘acute but ill-defined CVD’; 437.0, ‘cerebral atherosclerosis’; 437.

No institution

except the Zurich centre offered structured programmes during the study period. Nearly all institutions reported providing – in addition to ‘standard care’ – ‘frequent short counselling’, half of the institutions reported offering ‘detailed counselling’ if indicated, and around half reported handing out information booklets. Also, institutions reported using nicotine substitution, or prescribing bupropion or varenicline in some patients. All institutions reported referring patients to specialized addiction treatment institutions if the patient so wished. During the intervention at the Zurich centre from November 2007 to December 2009, 1689 participants had 6068 cohort visits, and 46% smoked at their last visit (Table 1). Smoking status checklists were not available for 739 of 6068 visits (12%) and incomplete CX-5461 datasheet for 208 (3.4%), so that 5121 (84%) completed checklists were available. Visits with missing checklists were more likely to arise for nonsmoking participants (56%) than for currently smoking participants (44%). There was variation in the number of missing checklists between physicians (data not shown). Current smoking was

declared in 44.5% of the completed checklists. Among the 2374 checklists for those currently smoking, motivation was assessed as: 85 (3.6%) intended to stop immediately; 262 (11%) intended to stop within 6 months; 804 (33.9%) would stop later; 784 (33%) did not intend to stop; and 439 (18.5%) did not answer. Smoking cessation counselling was carried out in 1888 of 2374 visits (80%) for current smokers. Reasons for not counselling were: other priorities (50%), patient refusal (19%), lack of time (12%) find more and other reasons (18%). Among counselled participants, the following types of RNA Synthesis inhibitor additional support were given (multiple types per patient possible): distribution of handout (8.1%), detailed counselling (6.5%), varenicline prescription (3.8%), nicotine substitution

(2.5%), follow-up date arranged (2.4%), agreed upon stop date (1.5%), bupropion prescription (0.9%), and referral to specialized institution (0.2%). Changes in motivation were very common (Table 2), with the exception of persons who did not smoke, of whom 95% remained nonsmokers. In smokers, the probability of a change in motivation level between two visits was more than 50% (diagonal elements in Table 2). The probability of changing from smoking to not smoking between two visits strongly depended on the motivation level, with 14% among persons ready for an immediate stop and 13% among those intending to stop within the next 6 months, but only 5.3% for persons who indicated to stop later, and 5.1% for those who were not motivated at all. When compared with ‘no motivation’, the odds ratios (95% confidence intervals) for not smoking at the next visit were 1.9 (0.85–4.2) for ‘immediate stop’, 2.1 (1.2–3.8) for ‘stop within 6 months’, and 1.0 (0.61–1.7) for ‘stop later’.

, 2006; Sisto et al., 2009; Fujimoto et al., 2011). Two primer sets have hitherto been reported as L. rhamnosus GG-specific primer sets (Brandt & Alatossava, 2003; Ahlroos & Tynkkynen, 2009). However, few studies have used the strain-specific primer sets, and the qualities

of the sets remain to be characterized. In this study, the two published L. rhamnosus GG-specific primer sets were evaluated by focusing on strain specificities of the sets for future use. All strains used in this study are shown in Table 1. L. rhamnosus GG (=ATCC 53103) was obtained from the American Type Culture Collection and used as positive control. L. rhamnosus DSM 20021T was from the German Collection of Microorganisms and Cell Cultures and www.selleckchem.com/products/PD-0332991.html used as negative control. A number of dairy isolates and human clinical isolates originating from different countries and identified as Venetoclax clinical trial L. rhamnosus were obtained from the Belgian Coordinated Collection of Microorganisms/LMG. The strains were cultured

in MRS broth at 37 °C for 20 h. Bacterial DNA was extracted from 1 mL of the cultured cells as previously described (Endo et al., 2007). Two different L. rhamnosus GG strain-specific PCR systems were used in this study, and all PCR primers used are shown in supporting information Table S1. The first PCR system targets a putative transposase gene in L. rhamnosus GG as described by Ahlroos & Tynkkynen (2009). Preparation of the reaction mixture and amplification of DNA

were conducted as described by Ahlroos & Tynkkynen (2009). The second PCR system targets a phage-related gene in L. rhamnosus GG as described by Brandt & Alatossava (2003). Preparation of the reaction mixture and amplification of DNA were according to a method previously described (Brandt & Alatossava, 2003). The amplification products were subjected to gel electrophoresis in 1.0% agarose, followed by ethidium bromide staining. Rep-PCR, RAPD, and ERIC PCR fingerprinting 3-mercaptopyruvate sulfurtransferase were carried out for strain differentiation in L. rhamnosus strains. (GTG)5 primer and a primer set REP1R-I and REP2-I were used for rep-PCR (Table S1). Preparation of the reaction mixture and amplification of DNA were according to the method described by Gevers et al. (2001). For RAPD fingerprinting, six different primers (C0540, 1251, OPA-03, D, E, and F) were used (Table S1). Preparation of the reaction mixture and amplification of DNA were performed as described elsewhere (Endo & Okada, 2006). PCR primers ERIC-1 and ERIC-2 were used for the ERIC PCR (Table S1). Preparation of the reaction mixture and amplification of DNA were by the method of Ventura et al. (2003). The amplification products were subjected to gel electrophoresis in 1.0% agarose, followed by ethidium bromide staining.