The compactin-producing strain P solitum 20-01 was obtained from

The compactin-producing strain P. solitum 20-01 was obtained from the laboratory collection of Centre ‘Bioengineering’ RAS. Culture conditions and harvesting of mycelia were as described before (Dzhavakhiya & Voinova, 2006).

Mycelia were freeze-dried and ground to fine powder. Total DNA was isolated using DNeasy Plant Mini Kit (Qaigen) according to the manufacturer’s instructions. Mitochondrial genome sequencing was performed using a total genomic DNA sample without prior isolation of the mitochondrial DNA. The genome was sequenced on a Roche Genome Sequencer Z-VAD-FMK purchase FLX using Titanium protocol for a shotgun genome library. The GS FLX run resulted in the generation of about 470 MB of sequences of an average read length of 379 bp. The GS FLX reads were assembled into contigs using the ‘GS de novo assembler’. Sequence coverage was 22×. A single 28 601-bp contig was identified as representing the mtDNA on the basis of extensive sequence similarity to known yeast mitochondrial genomes. MFannot tool ( with default settings was used for mitochondrial genome annotation, which was manually adjusted by sequence alignment of deduced genes with their intronless orthologs from related species. Putative proteins encoded by gene

models with no similarity to characterized genes were analysed by blast homology search against NCBI protein database The codon frequency ABT-888 in vivo was determined with CodonW (Peden, 2005) for concatenated ORFs for all protein-coding genes in the P. solitum mitochondrial genome. Genome contigs, corresponding to P. chrysogenum, A. oryzae and A. terreus mitochondrial genomes, all contained ‘extra’ sequences that were actually duplications of a region of rnL gene and adjacent tRNA gene cluster. These ‘extra’ sequences were considered as assembly artefacts and were manually deleted in the course of annotation. The complete mtDNA sequence of P. solitum 20-01 mtDNA is available in GenBank PLEKHB2 (JN696111,

BioProject ID: PRJNA72889). Whole genome DNA comparison was performed using megablast (Altschul et al., 1997) against NCBI database and mVISTA genome visualization and comparison tool (Frazer et al., 2004). Genome visualization, search for conserved sequence motifs and DNA repeats were performed with Vector NTI (Lu & Moriyama, 2004) and Ugene ( For phylogenetic analysis, 14 mitochondrial proteins, including subunits of the respiratory chain complexes (cox1-cox3, cob), ATPase subunits (atp6, atp8 and atp9), and seven NADH:quinone reductase subunits (nad1, nad2, nad3, nad4, nad4L, nad5 and nad6), were concatenated and aligned using the MUSCLE algorithm included in the mega5 package (Tamura et al., 2011). The sequence data for 25 filamentous fungi and yeast species with complete mitochondrial genomes were used as follows: Arthroderma obtusum (FJ385029), A. oryzae (AP007176), Aspergillus tubingensis (DQ217399), A. terreus (AAJN01000268.

Aim  The aim of this study was to evaluate soda, juice, sugared-

Aim.  The aim of this study was to evaluate soda, juice, sugared-beverage intake, brushing habits, and community water source availability as they relate to the prevalence of both noncavitated and cavitated caries lesions

in small rural villages in Mexico. Design.  The International Caries Detection and Assessment System (ICDAS) was used in children from small, isolated, villages in Mexico. Risk factors were assessed via questionnaires. Results.  Caries prevalence in the villages was very high, ranging from 94.7% to 100% of the children studied. The mean number of surfaces with lesions per child (D1MFS + d1mfs) having scores ≥1 (noncavitated and cavitated) ranged from 15.4 ± 11.1 to 26.6 ± 15.2. Many of the children reported drinking beverages check details containing

sugar. Conclusions.  Drinking sugared beverages, poor oral hygiene habits, and lack of access to tap water were identified as risk factor for caries in this sample of residents of rural Mexico. “
“International Journal of Paediatric Dentistry RGFP966 nmr 2011; 21: 241–248 Objective.  The aim of this study was to clinically assess the effectiveness of masking white spot enamel lesions using a resin infiltration technique that was recently developed to arrest incipient caries in a micro-invasive concept. Methods.  Twenty teeth with a Developmental Defect of Enamel (DDE) and 18 teeth with Post-orthodontic Decalcification (POD) were selected and treated with resin infiltration. Standardized photographs were taken before, immediately after, and 1 week after treatment and were analysed using image analysing software to calculate the ΔE values. The results were classified into three groups: completely masked, partially masked, and unchanged. Results.  Among the 20 teeth with DDE, five teeth (25%) were classified as completely masked, whereas seven

(35%) and eight teeth (40%) were partially masked and unchanged, respectively. Among the 18 teeth with POD, 11 teeth (61%) were completely masked, six teeth (33%) were partially Methisazone masked, and one tooth (6%) was unchanged. In some teeth, the result was more improved after 1 week than immediately after infiltration. Conclusion.  The masking effect was dramatic in some cases but not in others. The long-term colour stability of the result should be followed up through continuous clinical and scientific studies. “
“International Journal of Paediatric Dentistry 2013; 23: 125–130 Background.  Few prospective studies on the anxiety of children in the dental office have been published. Aims.  To monitor dental anxiety levels in children with and without previous experience with toothache over a period of six consecutive visits. Design.  A longitudinal study was carried out involving 167 children treated at a public dental service.

To separate these plasmids and to transfer them to a nonpathogeni

To separate these plasmids and to transfer them to a nonpathogenic host, in vitro transposition was performed with transposon EZ::TN , bearing a kanamycin resistance

gene. This resulted in the selection of three recombinant LBH589 chemical structure plasmids in E. coli NM522: pIGMS31KAN, pIGMS32KAN, and pIGRKKAN (Table 1). Purified DNA of these plasmids served as the templates for DNA sequencing reactions. The position of the transposon insertion site in the individual plasmids is shown in Fig. 1. The full nucleotide sequences of plasmids pIGMS31 (2520 bp), pIGRK (2348 bp), and pIGMS32 (9294 bp) were determined. Interestingly, the plasmids pIGMS31 and pIGRK were found to have a very low GC content (32.7% and 33.4%, respectively; Fig. 1), well below that of pIGMS32 (55.2%) or the total DNA of K. pneumoniae (57%; Fouts et al., 2008; Wu et al., 2009), which suggested the relatively recent acquisition of these replicons Everolimus concentration by HGT. Detailed sequence analysis identified a number of putative functional genetic modules in the plasmids: (1) a replication system (REP; in pIGRK, pIGMS31), (2) a system for mobilization

for conjugal transfer (MOB; in pIGMS31, pIGMS32), (3) a toxin–antitoxin system (TA) encoding a ParE family toxin (in pIGMS32; Jiang et al., 2002), and (4) a phenotypic module responsible for bacteriocin (cloacin) production (in pIGMS32; Fig. 1). Comparative sequence analysis (NCBI database) revealed that pIGMS32 is identical to a recently reported plasmid pCKO3 from Citrobacter koseri ATCC BAA-895 (accession no. CP000823). Moreover, it shows significant similarity to other ColE1-like plasmids, such as CloDF13 (Nijkamp et al., 1986), and to a much larger plasmid 15S (23.7 kb) from K. pneumoniae strain 15 (Gootz et al., 2009; Fig. 1c). The core region of 15S is 100% identical to pIGMS32, but the structure of this plasmid has been affected by insertions and deletions generated by two transposons containing antibiotic resistance genes (Fig. 1c). This analysis also identified plasmids related to pIGMS31 and pIGRK, containing homologous

REP or MOB systems see more (Fig. 1a and b), which indicated recombinational shuffling of the plasmid-encoded genetic modules. Comparative sequence analysis revealed that plasmids pIGMS31 and pIGRK carry related replication systems. Their predicted replication initiation proteins (ReppIGMS31 and ReppIGRK) exhibit 35% identity at the amino acid sequence level. ReppIGMS31 also shows local similarities (c. 45% identity) to Rep proteins encoded by plasmids residing in Pectobacterium atrosepticum, Salmonella enterica, and E. coli (all Gammaproteobacteria), while ReppIGRK is most similar (58% identity) to a replication protein of pHW126 from Rahnella genomospecies 3 (strain WMR126; Rozhon et al., 2010).

6 Three hundred microliters of 50 mM DMSO was placed in the bulb

6. Three hundred microliters of 50 mM DMSO was placed in the bulb of the side arm and was then used to initiate the reaction. The oxidation of MV was monitored by the decrease in A600 nm and the rate of oxidation was determined using the millimolar extinction coefficient of the reduced form, being 1.13 mM−1 cm−1 (Kelly & Wood, 1994). Cell-free extracts learn more prepared from H. sulfonivorans S1T grown heterotrophically

on dimethylsulfone were used as the positive control. ATP production experiments were performed essentially as described previously (Boden et al., 2010) using 1 mM DMS as an energy source in place of thiosulfate. The kinetic parameters derived from the growth of S. stellata in chemostat culture on fructose (12 mM) or succinate (2 mM) are given in Table 1. The maximum yield coefficient (Ymax) increased in the presence of DMS, which was oxidized stoichiometrically to DMSO SB431542 without assimilation into biomass. No DMS was detected in the cultures in a steady state. Upon the addition of DMS to a succinate or a fructose-limited chemostat, there was no immediate perturbation of the steady state and the dissolved oxygen concentration did not begin to decrease for approximately 6 h in the case of fructose or 3 h in the case of succinate, independent of the dilution rate.

The delay in oxygen consumption in the presence of DMS would indicate that the enzyme system for DMS oxidation was not constitutively expressed and the culture essentially underwent a lag phase while expression was induced. While the Ymax increased, it should be noted that the maintenance coefficient (mS) remained constant in the case of both carbon sources used. This was also the case when thiosulfate was used to support

the chemolithoheterotrophic growth of Methylophaga thiooxydans (Boden et al., 2010) and mixotrophic growth of Acidithiobacillus thiooxidans (Mason & Kelly, 1988). As stated previously, it is not possible to compare these data with those of Green et al. (2011) second owing to insufficient data being available from their paper to calculate Y– i.e. without quantifying substrate disappearance, Y cannot be calculated. The theoretical Ymax for growth on succinate is 37.1 g dry biomass mol−1 succinate (9.23 g dry biomass mol−1 substrate carbon), calculated using the assumption that 32% of succinate carbon is assimilated to biomass, as per the determinations performed by Anthony (1982) in a range of organisms. The experimental Ymax for succinate was found to be 33.6 g dry biomass mol−1 succinate (8.4 g dry biomass mol−1 substrate carbon), which increased in the presence of DMS to 38.9 g dry biomass mol−1 succinate (9.7 g dry biomass mol−1 substrate carbon) – this is higher than the theoretical Ymax and a 16% increase on the Ymax in the absence of DMS. The theoretical Ymax for growth on fructose dissimilated to 3-phosphoglycerate via the Entner–Doudoroff pathway is 73.7 g dry biomass mol−1 fructose (12.

57 (103 × 107 cells mL−1), 30 days after inoculation, and then t

57 (1.03 × 107 cells mL−1), 30 days after inoculation, and then the concentration declined rapidly (Fig. 1). The highest concentration of signaling molecules in the culture was about 18 nM relative to the reference OOHL based on the β-galactosidase activity. Mass scan analysis from 50 to 800 Da showed that three compounds in the metabolites of M. aeruginosa possessed the characteristic lactone moiety at m/z 102 of AHL-like molecules at retention time of 25.7, 27.7, and 39.2 min (Fig. 2). One of the compounds eluted at 39.2 min exhibited a quasi-molecular ion peak at m/z 256, in addition to the typical ion at m/z 101.8 that

is characteristic of an AHL fragment (Shaw et al., 1997). The ion at m/z 238 owing to [M +H−18]+ was produced by the AHLs because of the loss of water from the alkyl chain GSK126 price (Morin et al., 2003). These common features disclosed that this compound also belonged to the C4–14 AHL series. However, the strongest product of ions at m/z 88.1 is quite different from either the 3-oxo-C4–14 AHL compounds whose putative Ceritinib solubility dmso diagnostic ions

often appeared at m/z 98 (Ortori et al., 2007) or the 3-hydroxy-AHLs series whose diagnostic ions appeared at m/z values of 55, 69, 83, 97, etc., according to different alkyl chain length (Shaw et al., 1997). As for the unsubstituted acyl side chains systems, the diagnostic ions at m/z values of 95, 109, 123, and 137 become more prevalent (Ortori et al., 2007). This observation proved the existence of a CH3CH(OH)CH2CO-unit in the alkyl chain. Moreover, the quasi-molecular ion peak at m/z 256, along with the AHL moiety led to the deduction of the structure (Fig. 2). SEM photographs of M. aeruginosa showed that

the algal cells seemed to be experiencing free-living (< 20 day), aggregation (20–40 days), and disintegration (> 40 days) growth phases under laboratory culture conditions (Fig. 3). In addition, a biofilm-like membrane Liothyronine Sodium layer formed at 30 days after inoculation, which accompanied a strong aggregation of the cells (Fig. 3c1 and c2). To test the biological effects of QS signal, algal cells were cultured in BG-11 medium containing AHLs extracts (about 20 nM relative to the reference OOHL), which was obtained from the culture of M. aeruginosa at 30 days after inoculation. Compared with those in the fresh BG-11, the AHLs extracts could promote the formation of a biofilm-like membrane in M. aeruginosa, which appeared at 20 days (Fig. 3b2) and became thicker at 30 days (Fig. 3c2) after inoculation. QS that involves AHLs has been described in more than 70 different Gram-negative species of bacteria. All AHLs are composed of the conserved homoserine lactone ring and an amide (N)-linked acyl side chain that varies in the range of 4–18 carbons, may be saturated or unsaturated and be with or without the substitution at the third position (usually hydroxy- or oxo-) (Czajkowski & Jafra, 2009).

Whereas most studies have described cases of leptospirosis occurr

Whereas most studies have described cases of leptospirosis occurring during an outbreak, this study describes

a series of travel-related leptospirosis cases. Our cases were also confirmed by MAT that is not only sensitive and specific but also enables the determination of serogroups as it uses antigens of 17 different Leptospira serovars (Table 1). Nine different serovars were thus identified in this series. Travel represents the main sources of leptospirosis in nonendemic areas.[6, 11-14] In our experience (data not showed), 84% of the cases of leptospirosis diagnosed in the department in Paris between January 2008 and September 2011 were linked to travel in the tropics. In contrast, in endemic areas travel represents a less frequent but still significant source of leptospirosis. In Israel, a country endemic for leptospirosis, of 48 cases of leptospirosis diagnosed between 2002 and 2008, 42% were travel related.[7] In the Netherlands, LDE225 datasheet of 237 cases of leptospirosis diagnosed between 1987 and 1991, 14% were travel related.[6] In a western part of France also endemic for leptospirosis, of 34 patients seen over a period of 10 years, only 6% were

related to travel.[15] The most striking result was a history of at-risk exposure in Africa in 20% of our cases. Indeed, most travel-related leptospirosis cases have been described after travel to Asia, the Caribbean, and Central and South America.[4-6, 15] The incidence of leptospirosis in Africa is unknown. Travelers as epidemiological sentinel point toward this risk in Africa. An epidemic of leptospirosis in Kenya in 2004 also suggests that the disease is present but underdiagnosed.[16] Moreover, a study found a seroprevalence of 15% for leptospirosis in a population of five villages in the northeastern Gabon (Africa).[17] Overall, these recent studies clearly indicate that leptospirosis is underdiagnosed in Africa. Our epidemiological results are in agreement with those found in other series, with a predominance of males,[18]

at-risk fresh-water exposure such as bathing and practicing sports (canoeing, kayaking, rafting), together with a history of skin lesions[19] and high levels of hospitalization. In contrast with those studies, we did not find a predominance of the icterohemorragic serogroup but Diflunisal a large number of serogroups were involved. This indicates the wide diversity of the serogroups responsible for leptospirosis in travelers. The clinical picture is in agreement with that described in the literature with, in order of frequency, fever, headache, digestive disorders (vomiting, diarrhea, and nausea) myalgias, and arthralgias.[6, 7, 20] Laboratory results were also concordant with those found in the literature, with increased ASAT/ALAT, lymphocytopenia, thrombocytopenia, and renal impairment.[7, 12] We found a high frequency of lymphocytopenia (80%), a percentage higher than that usually reported in the literature, but similar to that found in another study.

, 2011) Together these data suggest that the role for thiamine i

, 2011). Together these data suggest that the role for thiamine in acid tolerance may well result from the requirement for acetoin production from pyruvate under conditions of acid stress, although further experiments will be required to test this model rigorously. The authors are grateful to members of the Bacterial

Stress Response Group at NUI Galway for helpful discussions and comments on the manuscript. We thank Prof Simon Foster for providing us with EGD (pLTV3). The work was supported by a Science Foundation Ireland SIRG award to K.A.K. (09/SIRG/B1570) and by an Irish Research Council for Science, Engineering and Technology EMBARK award to M.U. “
“A novel expression system for Lactobacillus plantarum was developed. This system is based on the manganese starvation-inducible promoter from specific manganese transporter of L. plantarum NC8, which was cloned for the first time. The Obeticholic Acid datasheet expression of a β-glucosidase from Pyrococcus furiosus (CelB) was achieved by cultivating L. plantarum NC8 at low manganese concentrations with MRS medium and the pmntH2-CelB expression vector. Adriamycin A CelB activity of 8.52 μkatoNPGal L−1 was produced in a bioreactor (4 L). The advantages of

the novel expression system are that no addition of an external inducing agent was required, and additionally, no further introduction of regulatory genes was necessary. The new promoter meets the general demands of a food-grade expression system. “
“Streptococcus pneumoniae is the main etiologic agent of pneumonia

worldwide. Because the members of the viridans group streptococci share a high degree of DNA sequence homologies, phenotypic and genotypic discriminations of S. pneumoniae from the viridans group are difficult. A quantitative real-time PCR assay targeting the capsular polysaccharide biosynthesis gene (cpsA) was developed as a species-specific detection tool for S. pneumoniae. The specificity was evaluated using genomic DNAs extracted from 135 oral cocci strains. Twenty-seven S. pneumoniae strains tested positive, whereas 108 other strains including Streptococcus pseudopneumoniae, Streptococcus mitis, and Streptococcus oralis did not show a specific signal. The linear regression of standard curves indicated high Afatinib correlations between the log numbers of S. pneumoniae cells and the CT values (R2=0.99). The minimal limit of detection was 32 fg of purified genomic DNA, equivalent to 14 genomes of S. pneumoniae. This new real-time PCR method may be very useful as a rapid and specific tool for detecting and quantifying S. pneumoniae. Potentially pathogenic Streptococcus pneumoniae, an α-hemolytic streptococci, is frequently detected in the oral environment with the viridans group streptococci, which constitute a major population of oral environments (Whatmore et al., 2000). Streptococcus pneumoniae causes pneumonia, otitis media, septicemia, and meningitis. Unfortunately, S.

The presence of five different plasmids in Sphingomonas sp MM-1

The presence of five different plasmids in Sphingomonas sp. MM-1 clearly demonstrated that there must

exists at Bortezomib mw least five different incompatibility groups in sphingomonads, and it can be assumed that the pronounced rearrangements, which occur after the conjugative transfer of degradative plasmids among sphingomonads, might be (at least in certain cases) related to incompatibility phenomena (Feng et al., 1997a, b; Ogram et al., 2000; Basta et al., 2004, 2005). The phenotypically defined incompatibility groups can be correlated with the sequences of the replication initiator (Rep) proteins and the proteins involved in plasmid partition (Par) (Petersen, 2011). In this context, the Rep proteins are especially important as these are responsible for the initial site specific DNA-binding and nicking activities, which represent the first steps in plasmid replication. The plasmid sequences deposited at the NCBI database originating from the genera Sphingomonas, Sphingobium, Novosphingobium and Sphingopyxis clearly demonstrated that the genes annotated as rep genes (repA or repB) almost exclusively belong to three protein superfamilies. Thus, proteins belonging to the RepA_C superfamily (Pfam 04796), Rep_3 (Pfam 01051) and

RPA superfamily (Pfam 10134) were found in large numbers among the deposited sequences (Table 1). In addition, the Rep proteins from four smaller plasmids (pUT2, pYAN-1, pSx-Qyy, Spl) Histone Methyltransferase inhibitor – which do not carry any catabolic genes – were classified to belong to the HTH-36 superfamily (Pfam 13730). An alignment of the Rep-sequences allowed the construction of a dendrogram to visualize the relationship among the different Rep-sequences (Fig. 1). This demonstrated that the Rep proteins from the large degradative plasmids

pNL1, pCAR3, pSWIT02 and Mpl can be clearly differentiated from the Rep proteins encoded by other plasmids. Thus, these Rep proteins belong to Methamphetamine the RepA_C family and are composed of about 430 amino acids (aa). In contrast, all other annotated Rep proteins are almost consistently smaller than 400 aa and did not belong to the RepA_C superfamily (Table 1). A second group of ‘megaplasmids’ consists of plasmid pISP1 (172 kbp) from Sphingomonas sp.MM-1, pNL2 (487 kbp) from N. aromaticivorans F199 and Lpl (192 kbp) from Novosphingobium sp. strain PP1Y. These plasmids encode for Rep proteins belonging to the RPA superfamily. Obviously, the plasmids of this group (=‘Mega-RPA’) are compatible with those of the group defined above (=‘Mega-RepAC’) as plasmids pNL1 and pNL2 are found together in N. aromaticivorans F199, and plasmids Mpl and Lpl in Novosphingobium sp. strain PP1Y (Romine et al., 1999; D’Argenio et al., 2011). A third group of large degradative plasmids was identified among the plasmids that possess a Rep protein belonging to the Rep_3 superfamily.

However, several variations in the life cycle occur among Ustilag

However, several variations in the life cycle occur among Ustilaginaceae

species. For instance, their systemic growth ability differs according to the plant organ infected (root, leaf or flower) (see Vánky, 1994). Among these variations, the role of solopathogenic strains was poorly investigated although such strains were considered as useful genetic tools. In the literature, solopathogenic strains of Ustilaginaceae were isolated from the progeny of in vitro mated haploid and compatible yeast strains, either wild types (Ehrlich, 1958; Puhalla, 1968) or auxotrophic mutants (Holliday, 1974; MEK inhibitor Harrison & Sherwood, 1994). Our strategy was to isolate solopathogenic strains from germinating teliospores to evaluate and compare the production of such spores under control conditions by three different smut fungi. In Ustilaginaceae,

dikaryotic cells are unstable and revert to haploid yeasts (Trueheart & Herskowitz, 1992), whereas solopathogenic strains are stable in axenic culture. RG7422 purchase This characteristic was used to eliminate fuzzy-dikaryotic strains from successive subcultures. Using this protocol, here we report the first isolation of a naturally occurring solopathogenic strain of S. reilianum, SRZS1. Nucleus staining revealed that SRZS1 is monokaryotic. The amplification and restriction enzyme digestion of mating type genes showed that SRZS1 has the two MATb alleles provided by the parental strains SRZM (MATb2) and SRZN (MATb1). This result is in agreement with the hypothesis that this monokaryotic strain is diploid. However, the strategy used did not allow us to exclude that the presence of the compatible

allele could also be the result of a parasexual transfer leading to the formation of a merodiploid strain (Zeigler et al., 1997), although such a mechanism has not been observed as yet in solopathogenic strains of U. maydis. Using specific primers of S. reilianum, PCR detection of SRZS1 in caulinar apices after crown infection showed that the strain is infectious. Its pathogenicity is weak as colonization did not lead to the formation of a sorus. We obtained similar conclusions with two other solopathogenic strains isolated from a poly-teliosporal sample (Table 1): inoculated plants present symptoms (dwarf Erythromycin plants and/or chlorotic spots on leaves), but did not develop smutted ears. Although they are infectious, the solopathogenic strains of S. reilianum seem unable to perform the entire life cycle of the fungus. We compared the ability of teliospores from M. penicillariae, S. reilianum and U. maydis to form solopathogenic cells. Surprisingly, all strains formed by the M. penicillariae were solopathogenic. It has already been described that monoisolates of this species can be infectious (Wilson & Bondari, 1990). Under our assay conditions, the solopathogenicity of monoisolates formed after teliospore germination is the usual cell status.

We collected information on HIV testing rates among MSM from 2001

We collected information on HIV testing rates among MSM from 2001 to 2011. Linear regression Epacadostat chemical structure was performed to estimate the change in HIV testing rates over time, with 95% confidence intervals (CIs), using information obtained from the available studies. Spearman’s rank correlation was performed to investigate the relationship between testing rates and the average age of surveyed MSM (P-value < 0.05 represents statistical significance). All analyses were performed in stata 10 (version 10.0, College Station, Texas, USA). We identified 1878 articles using the initial keywords (1872 articles were obtained from eight electronic databases and six relevant articles were

identified from the reference lists of these articles). After screening the titles of the 1878 articles, 1574 articles were excluded because of duplication or because they were unrelated to the topic. The abstracts of the remaining 304 articles were screened, and 97 articles were further excluded because they were not related to the topic. There were 207 articles eligible for full-text screening, of which 152 articles were subsequently excluded (143 articles did not report the level of HIV testing; five were duplicated in the databases;

two reported HIV testing rates among male sex workers, and two did not report the study period). Finally, we identified 55 eligible articles (44 in Chinese and 11 in English) that reported the HIV testing rate among Chinese MSM, Hydroxychloroquine which in total provided 37 testing rate estimates for individuals who had ever been tested for HIV during 2002–2009 and 29 testing rate estimates for individuals who had been tested in the past 12 months during 2003–2009 (Table 1). The selection process is illustrated in Figure S1. Among the 55 studies, eight studies reported the HIV testing rate in multiple years [25-32]. Eight of the 55 studies

did not report the recruitment method, while 24 studies recruited MSM participants from MSM venues, six recruited from Internet sites, five recruited from VCT clinics and one recruited Demeclocycline from MSM community settings; 12 studies used multiple recruitment methods (Table 1). The sample sizes of the studies ranged from 20 to 5454 [median 402; interquartile range (IQR) 202–558]. Our trend analysis across all available studies suggested that the percentage of MSM who had ever been tested for HIV increased from ∼10.8% (95% CI −2.8–24.4%) in 2002 to ∼51.2% (95% CI 39.0–63.4%) in 2009, with an average annual growth rate of ∼5.8% per year (95% CI 2.4–9.1%) (P = 0.0013) (Fig. 1a). The percentage of Chinese MSM who reported testing for HIV in the past 12 months also increased significantly, from ∼11.0% (95% CI −4.2–26.2%) in 2003 to ∼43.7% (95% CI 37.1–50.2%) in 2009, with an average increase of approximately 4.9% per year (95% CI 1.8–8.1%) (P = 0.0034) (Fig. 1b). Four of the 55 reported that approximately 82–97% of tested MSM were also notified about their HIV status after confirmation tests [25, 33-35].