, 2001 & Pillai et al., 2006). Like StcE, V. cholerae TagA is a secreted mucinase and contributes to colonization of the intestinal epithelium (Szabady et al., 2010). The A.  hydrophilia TagA exhibits an additional StcE function by cleaving and localizing C1-INH to the surface of bacterium, increasing the serum resistance check details of the bacterium in vitro. An isogenic deletion mutant of tagA

decreased the mortality of mice compared with wild-type A. hydrophila in a mouse model of peritonitis (Pillai et al., 2006). Thus, StcE-like metalloproteases play a role in the virulence phenotypes of A. hydrophila, V. cholerae and E. coli O157:H7. In this study, we identified stcE as a possible virulence factor in atypical Shigella B13 strains and further characterized this unique cluster of attaching and effacing pathogens. We would like to thank Thomas Whittam, Alison O’Brien, and Fred Blattner for bacterial strains, Entinostat manufacturer Nancy Strockbine for information regarding the atypical Shigella B13 strains, Jay Bangs for use of his epifluorescence microscope, and Rose Szabady

and Becca Moritz for insightful discussions regarding the project and critical reading of the manuscript. This work was supported by NIH grant RO1 AI051735. “
“Division of Natural Sciences and Mathematics, Transylvania University, Lexington, KY, USA Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae. Neisseria spp. have been shown to preferentially take up and transform their own DNA by recognizing a non-palindromic 10 or 12 nucleotide DNA uptake sequence (DUS10 or DUS12). We investigated the ability

of the DUS12 to enhance single-stranded DNA (ssDNA) transformation. Given the non-palindromic nature of the DUS12, we tested whether both strands of the DUS equally enhance transformation. Recombinant single-stranded M13 phage harboring transforming DNA with Y-27632 in vivo the Watson DUS12, the Crick DUS12, or no DUS (DUS0) were constructed and circular ssDNA was purified. Southern blots of the purified DNA probed with strand-specific oligonucleotide probes showed > 10 000 : 1 ratio of ssDNA to contaminating dsDNA. The Crick strand of the DUS12 enhanced ssDNA transformation 180- to 470-fold over DUS0 ssDNA, whereas the Watson strand of the DUS only modestly enhanced ssDNA transformation in two strains of N. gonorrhoeae. These data confirm that ssDNA efficiently transforms N. gonorrhoeae, but that there is a strand preference and that part of this strand preference is a greater efficiency of the Crick strand of the DUS12 in enhancing transformation. Natural transformation is a widespread mechanism for horizontal genetic exchange used by numerous bacterial species (Johnsborg et al.

Another genomic fragment containing sciP and adjacent ctrA was amplified using primers sciP-comF and ctrA-comR for disrupting both genes. The KIXX cartridge replaced a 644-bp SmaI/BamHI fragment, deleting the last 215 bp of sciP and the first 331 bp of the 711 bp ctrA. Plasmids containing disrupted versions of the genes were conjugated to R. capsulatus from E. coli C600 (pDPT51) (Taylor et al., 1983). Mutant strains were generated by GTA transfer of the disrupted versions of the genes into the chromosome of SB1003 (Scolnik & Haselkorn, 1984). PCR

with the original amplification primers was Linsitinib used to confirm the resulting kanamycin-resistant strains contained only the disrupted genes. Plasmid recombineering (Noll et al., 2009) was used for the generation of the cckA deletion construct. Primers cckA-p1 and cckA-p2 (Table 1)

were used to amplify a 4351-bp region encoding cckA (rcc01749) plus ~1 kb of flanking Metformin in vitro sequence on each side. Gel-purified PCR fragments were recombined into pUC19 (Vieira & Messing, 1982), and parental plasmids were selectively linearized by SalI treatment. Primers cckA-p3 and cckA-p4 (Table 1) were designed to PCR-amplify kanamycin resistance cassette A002 (Gene Bridges, Germany) with 50-bp tails homologous to cckA bases 53–103 and 2202–2252, respectively. λ-Red recombination resulted in the replacement of ~91% of plasmid-encoded cckA with the kanamycin resistance marker, yielding pUCΔcckA. The resulting plasmid was used to generate the cckA mutant strain as described above for the chpT, sciP and the ctrA/sciP double mutants.

Trans complementation of wild-type genes under control of their native upstream sequences was performed with the low copy number broad-host-range plasmid, pRK767 (Gill & Warren, 1988). The complementing fragments were amplified from the genome with appropriate primers (Table 1). Site-directed mutagenesis was performed with the QuikChange Lightning Site-Directed Mutagenesis Kit (Stratagene) to create pRK767-borne mutant ctrA genes with their native promoter region encoding a CtrA phosphomimetic protein, CtrAD51E (Domian et al., 1997), and a CtrA protein that is unable to be phosphorylated, CtrAD51A (Ryan et al., 2002), using primers D51E-F/D51E-R Amrubicin and D51A-F/D51A-R, respectively (Table 1). The mutagenesis created single bp substitutions that resulted in a glutamate (D51E) or alanine (D51A) in place of the conserved aspartate (D51) phosphorylation site; the presence of the mutations was confirmed by sequencing. These plasmids and the empty pRK767 control were transferred to R. capsulatus via conjugation using E. coli S17-1 (Simon et al., 1983). RcGTA packages random fragments of the R. capsulatus genome and transfers these to recipient cells. A gene transfer bioassay was used to measure production and release of RcGTA particles.

(2008) Curr. Biol., 18, 684–688). “
“A challenge for researchers in the time-perception Selleck 3-Methyladenine field is to determine whether temporal processing is governed by a central mechanism or by multiple mechanisms working in concert. Behavioral studies of parallel timing offer interesting insights into the

question, although the conclusions fail to converge. Most of these studies focus on the number-of-clocks issue, but the commonality of memory mechanisms involved in time processing is often neglected. The present experiment aims to address a straightforward question: do signals from different modalities marking time intervals share the same clock and/or the same memory resources? To this end, an interval reproduction task involving the parallel timing of two Sirolimus mouse sensory signals presented either in the same modality or in different modalities was conducted. The memory component was tested by manipulating the delay separating the presentation of the target intervals and the moment when the reproduction of one of these began. Results show that there is more variance when only visually marked intervals

are presented, and this effect is exacerbated with longer retention delays. Finally, when there is only one interval to process, encoding the interval with signals delivered from two modalities helps to reduce variance. Taken together, these results suggest that the hypothesis stating that there are sensory-specific clock components and memory mechanisms is viable. “
“Functional neuroimaging studies have implicated a number of brain regions, especially the posterior

parietal cortex (PPC), as being potentially important for visual–tactile multisensory integration. Neratinib purchase However, neuroimaging studies are correlational and do not prove the necessity of a region for the behavioral improvements that are the hallmark of multisensory integration. To remedy this knowledge gap, we interrupted activity in the PPC, near the junction of the anterior intraparietal sulcus and the postcentral sulcus, using MRI-guided transcranial magnetic stimulation (TMS) while subjects localized touches delivered to different fingers. As the touches were delivered, subjects viewed a congruent touch video, an incongruent touch video, or no video. Without TMS, a strong effect of multisensory integration was observed, with significantly better behavioral performance for discrimination of congruent multisensory touch than for unisensory touch alone. Incongruent multisensory touch produced a smaller improvement in behavioral performance. TMS of the PPC eliminated the behavioral advantage of both congruent and incongruent multisensory stimuli, reducing performance to unisensory levels. These results demonstrate a causal role for the PPC in visual–tactile multisensory integration. Taken together with converging evidence from other studies, these results support a model in which the PPC contains a map of space around the hand that receives input from both the visual and somatosensory modalities.

The role of this operon in Yersinia is unknown, although secondary structure prediction using the online software tool Phyre (Kelley & Sternberg, 2009) suggests that YPK_1206 is an IHF-like DNA bending protein (Fig.

S4). Although the amino acid sequences of these two proteins possess low similarity, their secondary structures share high similarity Talazoparib datasheet (Swinger & Rice, 2004). These analyses suggest that YPK_1206 may have roles in DNA bending and SraG may function as a regulatory element in this process. Comparative genomic analysis revealed that the YPK_1206 and YPK_1205 genes are only present in Y. pseudotuberculosis YpIII, Yersinia enterocolitica palearctica and Y. enterocolitica W22703, and YPK_1206 and YPK_1205 in YpIII share 90% similarity with Y. enterocolitica. The interaction region between YPK_1206 and SraG is conserved in both Y. enterocolitica Ibrutinib supplier strains, which suggests that SraG may be involved in YPK_1206-1205 operon regulation. Our results also suggest a role of SraG in YPK_1206-1205 mRNA

stability (Fig. 3 and Fig. S2), although further experiments are needed to prove this hypothesis. Our results also revealed that the coding sequence of YPK_1206 is necessary for SraG-mediated regulation, which suggests that SraG may negatively regulate the YPK_1206 mRNA via interaction with this region. This is similar to MicC-induced ompD mRNA regulation, which requires the C terminus of RNase E to be involved (Pfeiffer et al., 2009). RNase E has an established function in stable RNA, antisense RNA decay and sRNA-mediated regulation (Afonyushkin et al., 2005; Pfeiffer et al., 2009). Decreasing YPK_1206 Oxymatrine mRNA level by SraG may also rely upon RNase E, which needs to be further investigated. It has been shown that PNPase expression is post-transcriptionally regulated by affecting mRNA stability (Briani et al., 2008). The primary transcript of pnp is very efficiently processed by RNase III, which creates a structure

that is susceptible to specific recognition by PNPase, inducing its autocontrol (Briani et al., 2008). RNA structure prediction by MFOLD (Zuker, 2003) revealed a hairpin structure in the pnp mRNA leader sequence, which could be recognized by RNase III (data not shown). The hairpin region of pnp overlaps with the sraG gene, so deletion of the sraG gene may abrogate the hairpin structure and disrupt the autoregulation of pnp mRNA to increase the expression of PNPase. The effect of SraG on pnp mRNA is under investigation. Our proteomic analysis revealed that the mutant of sraG regulated expression of 16 proteins. Bioinformatic analysis demonstrated that there is no sequence similarity between those potential targets. However, three proteins are related to maltose metabolism and belong to two adjacent divergently transcribed operons. This suggests that SraG may be also involved in regulation of maltose metabolism.

Most pharmacist prescribers are active prescribers who perceive better patient management as a key benefit of their prescribing. Doctors who have worked with pharmacist prescribers and patients receiving care provided by a pharmacist prescriber are highly supportive and value their prescribing roles. Key themes generated from qualitative research were expertise in pharmacotherapy,

the quality of medicines related information and benefits for the wider healthcare team. Issues were, however, noted around a potential lack of continued funding and inadequate support networks. While acknowledging issues of recruitment, response and recall biases, positive patient attitudes were also a key finding of very recent survey based research. Attitudes were overwhelmingly positive with the vast majority agreeing/strongly agreeing that they were

totally satisfied with their consultation selleck and confident that their pharmacist prescribed as safely as their General Practitioner. Pharmacist prescribers were considered approachable and thorough, and most would recommend consulting a pharmacist prescriber. A slightly smaller majority would prefer to consult their General Practitioner if they thought their condition was getting worse and a small minority felt that there had been insufficient privacy and time for all their queries to be answered. One key limitation was the lack of engagement of pharmacist prescribers HDAC inhibitor in the research. Research of the awareness, views and attitudes of members of the Scottish general public towards non-medical prescribing found that more

than half of the respondents were aware of non-medical prescribing. A higher proportion was more comfortable with prescribing by pharmacists and nurses than other health professionals. Several issues relating to aspects of clinical governance were highlighted, specifically education of non-medical prescribers and protection of patient data. Evidence Non-specific serine/threonine protein kinase from the medical literature has demonstrated the importance of the consultation on patient outcomes and hence we have also focused in this area. We have developed and validated an assessment tool, based on the ‘Royal College of General Practitioners’ (RCGP) Video Assessment Tool’, for assessment of pharmacist prescribers’ consultation skills. The RCGP tool was modified to the ‘Pharmacist Consultation Assessment Tool’ (PharmaCAT). Competency areas of the RCGP tool were left unchanged but performance criteria for each were modified to reflect pharmacist prescribing. The PharmaCAT has been tested in the pharmacist prescriber setting. The tool had discriminatory power across different domains and inter-rater reliability. The PharmaCAT has potential to be used as a formative and/or summative assessment tool. Further research and developments in this field are being undertaken in collaboration with NHS Education for Scotland; an online version of PharmaCAT is being piloted.

“
“A major dose-limiting side effect of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) chemotherapies, such as the nucleoside reverse transcriptase inhibitors (NRTIs), is a small-fiber painful peripheral neuropathy, mediated by its mitochondrial toxicity. Co-morbid conditions

may also contribute to this dose-limiting effect of HIV/AIDS treatment. Alcohol abuse, which alone also produces painful neuropathy, is one of the most important co-morbid risk factors for peripheral neuropathy in patients with HIV/AIDS. Despite the prevalence of this problem and its serious impact on the quality of life and continued Screening Library concentration therapy in HIV/AIDS patients, the mechanisms by which alcohol abuse exacerbates highly active

antiretroviral therapy (HAART)-induced neuropathic pain has not been demonstrated. In this study, performed in rats, we investigated the cellular mechanism by which consumed alcohol impacts antiretroviral-induced neuropathic pain. NRTI 2′,3′-dideoxycytidine (ddC; 50 mg/kg) neuropathy was mitochondrial-dependent and PKCε-independent, and alcohol-induced painful neuropathy was PKCε-dependent and mitochondrial-independent. At low doses, ddC (5 mg/kg) and alcohol (6.5% ethanol diet for 1 week), which alone do not affect nociception, together produce profound mechanical hyperalgesia. This hyperalgesia is mitochondrial-dependent but PKCε-independent. These www.selleckchem.com/products/RO4929097.html experiments, which provide the first model for studying the impact of co-morbidity in painful neuropathy, support the clinical impression that alcohol consumption enhances HIV/AIDS therapy neuropathy, and

provide evidence for a role of mitochondrial mechanisms underlying this interaction. CYTH4 “
“The mechanisms that underlie the selection of an inhibitory GABAergic axon’s postsynaptic targets and the formation of the first contacts are currently unknown. To determine whether expression of GABAA receptors (GABAARs) themselves – the essential functional postsynaptic components of GABAergic synapses – can be sufficient to initiate formation of synaptic contacts, a novel co-culture system was devised. In this system, the presynaptic GABAergic axons originated from embryonic rat basal ganglia medium spiny neurones, whereas their most prevalent postsynaptic targets, i.e. α1/β2/γ2-GABAARs, were expressed constitutively in a stably transfected human embryonic kidney 293 (HEK293) cell line. The first synapse-like contacts in these co-cultures were detected by colocalization of presynaptic and postsynaptic markers within 2 h. The number of contacts reached a plateau at 24 h.

We identified three essential conserved residues (H204, Y236 and C266) that are critical for the assembly of type 1 fimbriae in this organism. rapid

amplification of cDNA ends analyses and reverse transcriptase-PCR results indicate that srtC1 was transcribed together with the putative adhesin gene fimQ and major structural subunit gene fimP as a single polycistronic mRNA. Actinomyces oris T14V (Henssge et al., 2009), formerly known as Actinomyces naeslundii T14V, a member of A. naeslundii genospecies 2 family, is considered as one of the primary colonizers for the formation of dental plaque on tooth surfaces (Li et al., 2004). Actinomyces oris T14V possesses two immunologically distinct types of fimbriae, which mediate the attachment of this species to both hard and soft HER2 inhibitor tissue surfaces (Cisar et al., 1988). These fimbriae were among one of the first observed in gram-positive bacteria (Girard & Jacius, 1974). Type 1 fimbriae promote the binding of this organism to tooth surfaces mediated

by the adsorbed salivary acidic proline-rich proteins and statherin. These salivary proteins serve as receptors for type 1 fimbriae (Clark VE-821 mouse et al., 1984; Gibbons et al., 1988). Type 2 fimbriae mediate the adherence of A. oris to oral mucosal epithelial cells and lactose-sensitive coaggregations with certain oral streptococci. Such interactions with other bacteria further promote the formation of dental plaque initiated by type 1 fimbriae of the organism (Palmer et al., 2003). Previously, we demonstrated that the biogenesis of functional type 1 fimbriae in A. oris T14V required three genes (Yeung et al., 1987; Chen et al., 2007): the putative adhesin gene fimQ, the major structural subunit gene fimP and the type 1 fimbria-specific sortase gene srtC1. Sequence alignment indicates that A. oris SrtC1 contains Cell press all three conserved domains (D1, D2 and D3) that are present in all sortases and an extra C-terminal hydrophobic domain. According to the sortase classification (Dramsi et al.,

2005), SrtC1 belongs to class C sortase family. Sortases are a group of bacterial thiol transpeptidases responsible for the covalent attachment of specific surface proteins to the cell wall envelope of gram-positive bacteria (Marraffini et al., 2006). These enzymes are involved in the expression of several virulence factors and the assembly of fimbriae, and have been considered as a target of anti-infective therapy (Maresso & Schneewind, 2008). SrtC1 is required for both the assembly of type 1 fimbriae in A. oris T14V and its adherence to saliva-coated hydroxylapatite (Chen et al., 2007). Accordingly, preventing the formation of type 1 fimbriae in A. oris by inhibiting the function of this sortase may reduce the colonization of this organism and consequently the dental plaque formation.

Some diseases, like chikungunya[10] tend to go undiagnosed. Our first two cases were initially suspected as having

dengue, both returning from Southeast Asia with fever followed by rash and thrombocytopenia, even leucopenia (Table 1). It is not an easy task to clinically distinguish the appearance of rash associated with each of the agents causing fever and skin manifestations. The endeavor becomes especially demanding, Obeticholic Acid research buy if a clinician is presented with a disease he or she has never come across—as is often the case for measles in Finland and Estonia. Measles is highly contagious. It is transmissible from 4 days before to 4 days after the onset of the rash. When misdiagnosed, the isolation of the patient is delayed, which allows more time for transmission. Hence, it is of particular importance for the doctors to be able to raise a suspicion of measles—leading to infection control measures without delay. The infection control measures taken for the present cases have been described elsewhere.[11] Notably, one of our patients had a short diarrhea, two were suspected as having mild pneumonia and urinary tract infection. www.selleckchem.com/ATM.html Approximately 30% of measles cases have one or more complications,[4] such as diarrhea (8%), otitis media (7%), pneumonia (viral or bacterial) (6%), and acute encephalitis (0.1%).[4] Bacterial superinfections appear to

be secondary to local tissue damage and depression of cellular immunity.[2] Travelers Methane monooxygenase may occasionally act as sentinels for ongoing outbreaks in their destinations. Our cases were reported on the European Network for Tropical Medicine and Travel Health (TropNet) member site, and we learned that no outbreaks of measles have been identified in Thailand as yet (Dr Jiri Beran, personal communication). While both flights with measles patients were charter flights flying non-stop from Phuket to Helsinki, transmission from other international travelers visiting Phuket remains

a possibility. However, our patients all stayed at different hotels and, moreover, the genotype of the virus in cases 1 and 2 was defined, and proved to be D8 known to be currently circulating in Thailand (MeaNS, http://www.who-measles.org). In Finland, with no autochtonous measles, all cases have originated in international travel. Out of the 20 measles cases identified among travelers since 1996, in 12 the disease was contracted in other European countries.[11] Notably, in countries where the disease is still encountered, like France, a short-term traveler with measles may have caught the disease already at home before departure.[12] Measles should be suspected as a cause of febrile rash in travelers returning from any area, even the most popular vacation resorts, such as Thailand.

Hinjiranandana (Somdej Pranangchao Sirikit Hospital, Chonburi); P. Layangool (Bhumibol Adulyadej Hospital, Bangkok); N. Kamonpakorn (Somdej Prapinklao Hospital, Bangkok); S. Buranabanjasatean (Mae Chan Hospital, Chiang Rai); C. Ngampiyaskul (Prapokklao Provincial Hospital, Chantaburi); T. Chotpitayasunondh, S. Chanpradub and P. Leawsrisuk (Queen Sirikit National Institute of Protease Inhibitor Library supplier Child Health, Bangkok); S. Chearskul, N. Vanprapar, W. Phongsamart, K. Lapphra, P. Chearskul, O. Wittawatmongkol, W. Prasitsuebsai, K. Intalapaporn, N. Kongstan,

N. Pannin, A. Maleesatharn and B. Khumcha (Department of Pediatrics, Faculty of Medicine, Siriraj Hospital, Mahidol University); L. Aurpibul, N. Wongnum and R. Nadsasarn [Research Institute for Health Sciences (RIHES), Chiang Mai University, Chiang Mai]; P. Lumbiganon, P. Tharnprisan and T. Udompanich (Department of Pediatrics, Faculty of Medicine, Khon Kaen University); M. Yentang (Petchburi Hospital, Petchburi); A. Khonponoi, N. Maneerat, S. Denjunta, S. Watanaporn, C. Yodsuwan, W. Srisuk, Volasertib concentration S. Somsri and K. Surapanichadul (Chiang Rai Regional Hospital, Chiang Rai). The authors would like to acknowledge

Dr. Nneka Edwards-Jackson for her help with manuscript preparation. “
“The aim of the study was to explore the awareness of rectal microbicides, the use of pre-exposure prophylaxis (PREP) and the willingness to participate in biomedical HIV prevention trials in a cohort of HIV-negative gay men. In a community-based cohort study, HIV-negative homosexually active men in Sydney, Australia were questioned about awareness of rectal microbicides, use of PREP, and willingness to participate

in trials of such products. Predictors of awareness and willingness to participate were analysed by logistic regression. Use of PREP was examined prospectively. Overall, 14% had heard of rectal microbicides. Older (P=0.05) and 4��8C university-educated men (P=0.001) were more likely to have knowledge of rectal microbicides. Almost one-quarter (24%) of men reported that they were likely/very likely to participate in rectal microbicide trials. Among those men with definite opinions on participation, awareness of rectal microbicides was significantly associated with unwillingness to participate [odds ratio (OR) 0.78, 95% confidence interval (CI) 0.65–0.93, P=0.007]. Willingness to participate in trials using antiretroviral drugs (ARVs) to prevent HIV infection was reported by 43% of men, and was higher among those who reported unprotected anal intercourse (UAI) with HIV-positive partners (OR 1.88, 95% CI 0.99–3.56).

8%) were lost to follow-up. The mean age of participants at follow-up was 27.1 years (SD 6.1 years) (compared with 26 years at baseline; SD 6.5 years) and HIV prevalence was 35.3% (78 learn more of 221). Among those who had received their serostatus 1 year before, a majority reported having disclosed their serostatus following

VCT (178 of 198; 89.9%) (Table 3). Of the 20 women who had not revealed their status, seven (35%) feared harassment or banishment by family, while 13 (65%) declared that one’s serostatus is private and thus does not have to be revealed. Seronegative women at follow-up were more likely to report status disclosure than seropositive women (93.8% vs. 82.4%, respectively; P=0.011). Serostatus (negative or positive) was generally revealed in the work environment, to other FSWs (56.2% of cases) or to worksite managers or owners (53.3%). Disclosure to significant others or health professionals occurred less frequently: ABT 263 29.8% reported disclosure to a regular partner, 19.7% to

the family and only 8.4% to a health agent (Table 3 reasons for disclosure included to receive moral support (52.2%), to encourage other people to be tested (29.2%) or to strengthen the relationship with their partner (12.4%). Other reasons for disclosure were also reported. Three participants (1.7%) reported having been forced to reveal their serostatus in order to be able to continue practising sex work at their worksite. Moreover, qualitative data collection confirmed these results by

showing that women who disclosed their serostatus at their worksites increased the pressure for disclosure on women who would not have otherwise disclosed their serostatus. Seronegative FSWs tended to disclose their status spontaneously and publicly, leading to suspicion of HIV seropositivity for women who chose to remain silent. Some sex workers reported that some peers revealed friends’ status to be detrimental to them. Qualitative data also confirmed that certain managers or owners of sites asked FSWs to disclose their serostatus if they were to continue to work at their sites. These managers wanted to be able to assure their customers of the ‘safety’ of their bars. Among disclosers, most (89.3%) reported receiving very positive reactions from the people to whom they disclosed their serostatus (Table 3). These positive reactions included moral Cepharanthine support, access to treatment and reinforcement of the relationship with the FSW’s regular partner. In fact, a quarter of subjects with regular sexual partners at baseline (boyfriend or husband) (42 of 168; 25.0%) reported that their partner was tested for HIV after the FSW’s own VCT, and the partner later disclosed his serostatus to the FSW in most cases (38 of 42; 90.5%). A few participants (nine) sought and obtained medical care after VCT and two are now receiving ART (Table 3). Psychosocial assistance was also provided to six participants in the AHS and in other health centres.