Conserved hypothetic proteins were aligned to pfam database for p

Conserved hypothetic proteins were aligned to pfam database for putative functions. In the initial experiments, we observed that the biofilm formation of S. aureus NCTC8325, which is rsbU defective, on a polystyrene or a glass surface was obviously inhibited in dithiothreitol-supplemented TSB. We postulated that the sulfhydryl group may play a role in biofilm inhibition. Selleck Rucaparib As expected, replacing dithiothreitol with BME or cysteine led to a similar phenomenon (Fig. 1a). The minimal biofilm-inhibitive concentrations of dithiothreitol, BME and cysteine were determined later by static biofilm formation assays on 96-well microtiter plates. The amount of the biofilms formed decreased gradually as the concentrations

of the supplemented sulfhydryl compounds increased. For S. aureus NCTC8325 cells, 5 mM dithiothreitol, 10 mM cysteine or 20 mM BME reduced over 70% biofilm formation on the polystyrene surface (Fig. 1b). To verify whether the BTK inhibitor screening library phenomenon is strain specific, the biofilm-forming abilities of several S. aureus strains and one S. epidermidis strain were tested in the presence of sulfhydryl compounds (Fig. 2). All three tested sulfhydryl compounds, including dithiothreitol, BME and cysteine, reduced biofilm formation in these staphylococcal strains, although the susceptibility varied among the different strains. To explore whether sulfhydryl compound

would cause a growth inhibition on S. aureus, we determined the growth curves of S. aureus NCTC8325 cells in TSB, TSB supplemented with 10 mM dithiothreitol, TSB supplemented with 20 mM cysteine and TSB supplemented with 40 mM BME by measuring OD600 nm at different time points. No significant difference in the growth rate among the samples was observed (Fig. S1). The result indicated that the biofilm inhibition caused by thiols probably involved the switch of bacterial physiological states rather than mafosfamide the inhibition of bacterial growth. The first step in the formation of an S. aureus biofilm is adhering to the matrix surface. We investigated

the primary attachment ability of S. aureus NCTC8325 cells on 24-well polystyrene plates with or without the presence of thiols. However, no difference was observed in the primary attachment abilities of the bacterial cells in the control group and the sulfhydryl compound addition groups (Fig. 3). The production of PIA is a major event for biofilm maturation in S. aureus. The transcriptional level of icaADBC was investigated to find whether PIA synthesis was affected after treatment with thiols. Real-time reverse transcriptase-PCR showed that the mRNA levels of ica in the bacterial cells pretreated with 5 mM dithiothreitol, 10 mM cysteine or 20 mM BME for 30 min were significantly decreased compared with the control (Fig. 4a). In addition, extracellular PIA was also measured by the Elson–Morgan assay. An icaADBC deletion mutant of NCTC8325 was used as the negative control.

LtrB intron (Perutka et al, 2004) The simple probabilistic mode

LtrB intron (Perutka et al., 2004). The simple probabilistic model has some limitations and the database is not sufficiently large to reliably examine the complex interactions discussed previously. Thus, it CP-868596 mw is necessary to test each consecutive target site predicted by the computer algorithm for the identification of a successful intron integration site. This work was supported by the Korean Systems Biology Research Project

(20100002164) of the Ministry of Education, Science, and Technology (MEST). Further support by the World Class University Program (R32-2009-000-10142-0) through the National Research Foundation of Korea funded by the MEST is appreciated. “
“Volatiles produced by bacterial cultures are known to induce regulatory and metabolic alterations in nearby con-specific or heterospecific bacteria, resulting in phenotypic changes including acquisition of antibiotic resistance. We observed unhindered growth of ampicillin-sensitive Serratia rubidaea and S. marcescens on ampicillin-containing media, when exposed to volatiles produced by dense bacterial growth. However, this phenomenon appeared to result from pH increase in the medium caused by bacterial volatiles rather than alterations in the properties of the

bacterial cultures, as alkalization of ampicillin-containing culture media to pH 8.5 by ammonia or Tris exhibited the same effects, while pretreatment of bacterial cultures under the same conditions prior to antibiotic exposure did not increase ampicillin resistance. Ampicillin was readily inactivated at pH 8.5, suggesting HTS assay that observed bacterial growth results from metabolic alteration of the medium, rather than Molecular motor an active change in the target bacterial population (i.e. induction of resistance or tolerance). However, even such seemingly simple mechanism

may provide a biologically meaningful basis for protection against antibiotics in microbial communities growing on semi-solid media. “
“To the authors’ knowledge, most studies on biofilm formation have focused on bacteria and yeasts. So far, biofilm formation by fungal plant pathogen has not been reported. In this study, the biofilm-forming capacity of Fusarium oxysporum f. sp. cucumerinum was evaluated. For biofilm quantification, a colorimetric 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium-hydroxide (XTT) reduction assay was used to observe metabolic activity. Fluorescence and confocal scanning laser microscopy revealed that the biofilms have a highly heterogeneous architecture composed of robust hyphae and extracellular polysaccharide materials. Additionally, the influence of physical factors on F. oxysporum biofilm formation and the susceptibility of biofilms to environmental stress was investigated.

Further analysis demonstrated that students were more aware of th

Further analysis demonstrated that students were more aware of the difficulties which may pose as threats to the patient-practitioner relationship, ‘There was a guy who was answering the Morisky (8-item

Medication Adherence Scale), which one was it…do you take all your medicines, he sort of said yes and winked at us’, and perhaps, would now consider those aspects which may affect comprehension, ‘…wording was a little difficult for them’, and hinder effective communication, ‘Language barriers, sometimes I found it was hard to communicate with the elderly’. This study has demonstrated that MPharm student contact, prior to pre-registration training, with

sheltered housing residents to conduct domiciliary http://www.selleckchem.com/products/AZD0530.html medication reviews this website could be important to professional development since it allows students to expand their knowledge in the key area of understanding how older individuals manage their medicines. An awareness of the factors which can contribute to medication misadventure amongst patients and identification of appropriate strategies to mediate this risk are now requisite skills of newly qualified pharmacists. Thus development of early opportunities which allow students to experience contexts in which misadventure may occur may lead to an improvement in patient care. 1. George, J. Munro, K. McCaig, D. Stewart, D. Risk factors for medication misadventure among residents in sheltered housing complexes. British Journal of Clinical Pharmacology. 2007; 63: 171–176. 2. Parsell, G. Gibbs, T. and Bligh, J. Three visual techniques to enhance interprofessional learning. Postgraduate Medical Journal. 1998: 74: 387–390. “
“Deborah Brooks, Ashleigh Hopps, Simon White Keele University, Staffordshire, UK This study took a qualitative approach to exploring the perspectives of community pharmacy staff on the Healthy Living Pharmacy (HLP) initiative Endonuclease Benefits were reported for

customers and staff and local communities, despite most customers appearing unaware of the pharmacy’s HLP status The findings support continued national roll-out of the initiative The HLP initiative has led to quality and productivity improvements in community pharmacy, and better public access to health and wellbeing services.1 Key features include quality and performance criteria for services provided and trained Healthy Living Champions (HLCs) in each HLP.1 Local Pathfinders are helping to establish how HLPs can best support people to change their lifestyles, improve their health and wellbeing and potentially health outcomes, as well as building the evidence-base for pharmacy’s contribution to public health.

Further analysis demonstrated that students were more aware of th

Further analysis demonstrated that students were more aware of the difficulties which may pose as threats to the patient-practitioner relationship, ‘There was a guy who was answering the Morisky (8-item

Medication Adherence Scale), which one was it…do you take all your medicines, he sort of said yes and winked at us’, and perhaps, would now consider those aspects which may affect comprehension, ‘…wording was a little difficult for them’, and hinder effective communication, ‘Language barriers, sometimes I found it was hard to communicate with the elderly’. This study has demonstrated that MPharm student contact, prior to pre-registration training, with

sheltered housing residents to conduct domiciliary Sirolimus medication reviews Selleck AZD5363 could be important to professional development since it allows students to expand their knowledge in the key area of understanding how older individuals manage their medicines. An awareness of the factors which can contribute to medication misadventure amongst patients and identification of appropriate strategies to mediate this risk are now requisite skills of newly qualified pharmacists. Thus development of early opportunities which allow students to experience contexts in which misadventure may occur may lead to an improvement in patient care. 1. George, J. Munro, K. McCaig, D. Stewart, D. Risk factors for medication misadventure among residents in sheltered housing complexes. British Journal of Clinical Pharmacology. 2007; 63: 171–176. 2. Parsell, G. Gibbs, T. and Bligh, J. Three visual techniques to enhance interprofessional learning. Postgraduate Medical Journal. 1998: 74: 387–390. “
“Deborah Brooks, Ashleigh Hopps, Simon White Keele University, Staffordshire, UK This study took a qualitative approach to exploring the perspectives of community pharmacy staff on the Healthy Living Pharmacy (HLP) initiative pheromone Benefits were reported for

customers and staff and local communities, despite most customers appearing unaware of the pharmacy’s HLP status The findings support continued national roll-out of the initiative The HLP initiative has led to quality and productivity improvements in community pharmacy, and better public access to health and wellbeing services.1 Key features include quality and performance criteria for services provided and trained Healthy Living Champions (HLCs) in each HLP.1 Local Pathfinders are helping to establish how HLPs can best support people to change their lifestyles, improve their health and wellbeing and potentially health outcomes, as well as building the evidence-base for pharmacy’s contribution to public health.

[10-12] College freshmen living in dormitories are at particularl

[10-12] College freshmen living in dormitories are at particularly high risk for developing meningococcal disease.[13] Because of this, students from overseas who are planning to live in college dormitories

are usually required to provide proof of meningococcal immunization in the United States and other countries such as the United Kingdom. Many Taiwanese students preparing to study in the United States are required selleck kinase inhibitor to have the vaccination, which is not a routine immunization in Taiwan.[14] In addition, the vaccine is available only at 12 Centers for Disease Control contracted hospitals due to the scarceness of the vaccine in Taiwan. However, receiving vaccination without learning about the disease is not enough to assure prevention and patient-level factors may influence immunization coverage. Furthermore, educating patients about the selleck products risk of contracting the disease and the importance of the vaccine

should be an essential part of the physician–patient discussion about vaccination. Thus far, few studies have investigated the awareness and attitudes toward meningococcal disease among high-risk students. We designed a study to survey the knowledge, attitudes, and behaviors about the disease among Taiwanese students planning to study in the United States. A cross-sectional questionnaire survey on Taiwanese college students planning to study in the United States was conducted in National Taiwan University Hospital in Taipei, a medical center-based travel medicine clinic, from January 2009 to December 2010. The questionnaire and consent forms were distributed to all college-age nonmedical students from different universities planning to study in the United

States. All study procedures were approved by the ethical committee of Meloxicam the National Taiwan University Hospital. A self-administered, single-choice questionnaire surveyed the background information, attitudes toward, and knowledge about meningococcal disease. The questionnaire was based upon personal practice experiences and designed after a careful literature review. Excluding background information, the questionnaire included two questions about attitudes, five questions about general knowledge of the disease, four questions on preventive or postexposure management, and two questions on individual preventive practices. Five experts tested the content validity, while the face validity was tested by five college students. Data management and statistical analyses were performed using SPSS 11.0 software. Frequency distributions were used to describe the demographic data. Stepwise logistic regression analysis determined the relative values of the variables related to positive attitudes on receiving vaccines and willingness to perform individual preventive practices. A p-value less than 0.05 was considered statistically significant.

We used whole-cell patch-clamp recording to measure glutamatergic

We used whole-cell patch-clamp recording to measure glutamatergic transmission in the dorsomedial striatum (DMS) and dorsolateral striatum (DLS). Ube3am−/p+ mice were severely impaired in initial acquisition of lever pressing. Whereas the lever pressing of wild-type controls was reduced by outcome devaluation and instrumental contingency reversal, the performance of Ube3am−/p+ mice were more habitual, impervious to changes in outcome value and action–outcome contingency. In the DMS, but not the DLS, Ube3am−/p+ mice showed reduced amplitude and frequency of miniature excitatory postsynaptic currents.

These results show for the first time a selective deficit in instrumental conditioning in the Ube3a deficient mouse model, and suggest a specific impairment

in glutmatergic transmission in the associative corticostriatal circuit Idelalisib in AS. “
“In synaesthetes, stimulation of one sensory pathway provokes a sensory experience (e.g. a colour concurrent) in a different sensory modality or sub-modality. Results of synaesthetic Stroop and priming tests indicate that the perception of a colour concurrent interferes with the processing of a veridical colour in synaesthetes. We here examined the congruency between a stimulus’ colour and the colour concurrent both in grapheme–colour synaesthetes and in non-synaesthetes trained on grapheme–colour associations. Electrophysiological (electroencephalogram) and behavioural measurements were collected HSP tumor during a priming task that included grapheme–grapheme and grapheme–colour patch pairs. To investigate covert bidirectional synaesthesia, an additional inverted colour patch–grapheme condition was included. Both groups of participants

showed longer reaction time and more negative-going N300 and N400 event-related potential (ERP) components on incongruent trials. Whereas ERP effects in the non-synaesthetes were Gefitinib datasheet largely confined to the late cognitive components N300, P300 and N400, the synaesthetes also showed congruency-dependent modulation of the early sensory component N170. Our results suggest that early cognitive processes distinguish cross-modal synaesthetic perceptions from acquired associations. The involvement of both early- and late-stage cognitive components in bidirectional synaesthesia possibly indicates similar feature-binding mechanisms during processing of opposite flow directions of information, namely grapheme–colour and colour–grapheme. “
“Matrix metalloproteinases (MMPs) are fine modulators of brain plasticity and pathophysiology. The inhibition of MMPs shortly after ischaemic stroke reduces the infarct size and has beneficial effects on post-stroke behavioural recovery. Our previous studies have shown that photothrombotic cortical stroke disrupts use-dependent plasticity in the neighbouring cortex. The aim of the present study was to check whether the inhibition of MMPs after photothrombosis rescued the plastic capacity of the barrel cortex.

The initial study was funded by the European Union contract no Q

The initial study was funded by the European Union contract no. QLK1-CT-2001-01066. We thank Dr J. Londesborough, VTT Biotechnology, Finland, for strain A15. Fig. S1. Alignment of the predicted proteins of maltose permease genes using clustalw. Fig. S2. Alignment of promoter sequences of maltose permease genes using clustalw. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“The Writing Group thanks the BHIVA Secretariat

for administrative help, Alison Richards for conducting the systematic literature search and Jacoby Patterson for work on critical appraisal, evidence profiles and construction Ipilimumab price of GRADE tables. The Writing Group also thanks Professor Francois Raffi and Professor Jose Arribas for their peer GSK126 clinical trial review of the guidelines and Dr Annemiek de Ruiter and Dr Fiona Lyons for their peer review of the section

on women. Dr Ian Williams has received grant support from Gilead Sciences and Janssen-Cilag and his department has received grant support from Boehringer Ingelheim, Gilead Sciences and Janssen-Cilag. Dr Duncan Churchill has no conflicts of interest to declare. Professor Jane Anderson has received lecture fees from Abbott, Gilead and ViiV and consultancy fees from Abbott, Bristol-Myers Squibb and Gilead. Her department has received a research grant from Gilead. Professor Jose

Arribas has a financial interest/relationship or affiliation: Tibotec, Janssen, Abbott, BMS, Gilead Sciences, MSD, ViiV Healthcare. Dr Marta Boffito has received consultancy fees Interleukin-3 receptor and grant support from Bristol-Myers Squibb, GlaxoSmithKline, Merck Sharp and Dohme, Pfizer, Roche and Tibotec/Janssen. Professor Mark Bower has no conflicts of interest to declare. Mr Gus Cairns has no conflicts of interest to declare. Dr Kate Cwynarski has received lecture and consultancy fees from Pfizer and Roche. Dr Annemiek de Ruiter has received lecture and consultancy fees from Bristol-Myers Squibb and Gilead. Dr Simon Edwards has received lecture fees from ViiV and Janssen, and consultancy fees from Boehringer Ingelheim, Merck Sharp and Dohme and ViiV. Dr S Fidler has no conflicts of interest to declare. Dr Martin Fisher has received lecture fees from Abbott, Astellis, Bristol-Myers Squibb, Gilead and ViiV and he has received consultancy fees from Abbott, Bristol-Myers Squibb, Gilead, Janssen, Merck Sharp and Dohme and ViiV. Dr Andrew Freedman has received lecture and consultancy fees from Abbott, Bristol-Myers Squibb, Gilead and Tibotec/Janssen. Professor Anna Maria Geretti has received consultancy fees from Gilead and her department has received research grants from Janssen, Merck Sharp and Dohme and ViiV. Dr Yvonne Gilleece has no conflicts of interest to declare. Professor Rob Horne has no conflicts of interest to declare.

The initial study was funded by the European Union contract no Q

The initial study was funded by the European Union contract no. QLK1-CT-2001-01066. We thank Dr J. Londesborough, VTT Biotechnology, Finland, for strain A15. Fig. S1. Alignment of the predicted proteins of maltose permease genes using clustalw. Fig. S2. Alignment of promoter sequences of maltose permease genes using clustalw. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“The Writing Group thanks the BHIVA Secretariat

for administrative help, Alison Richards for conducting the systematic literature search and Jacoby Patterson for work on critical appraisal, evidence profiles and construction see more of GRADE tables. The Writing Group also thanks Professor Francois Raffi and Professor Jose Arribas for their peer Omipalisib ic50 review of the guidelines and Dr Annemiek de Ruiter and Dr Fiona Lyons for their peer review of the section

on women. Dr Ian Williams has received grant support from Gilead Sciences and Janssen-Cilag and his department has received grant support from Boehringer Ingelheim, Gilead Sciences and Janssen-Cilag. Dr Duncan Churchill has no conflicts of interest to declare. Professor Jane Anderson has received lecture fees from Abbott, Gilead and ViiV and consultancy fees from Abbott, Bristol-Myers Squibb and Gilead. Her department has received a research grant from Gilead. Professor Jose

Arribas has a financial interest/relationship or affiliation: Tibotec, Janssen, Abbott, BMS, Gilead Sciences, MSD, ViiV Healthcare. Dr Marta Boffito has received consultancy fees Thiamet G and grant support from Bristol-Myers Squibb, GlaxoSmithKline, Merck Sharp and Dohme, Pfizer, Roche and Tibotec/Janssen. Professor Mark Bower has no conflicts of interest to declare. Mr Gus Cairns has no conflicts of interest to declare. Dr Kate Cwynarski has received lecture and consultancy fees from Pfizer and Roche. Dr Annemiek de Ruiter has received lecture and consultancy fees from Bristol-Myers Squibb and Gilead. Dr Simon Edwards has received lecture fees from ViiV and Janssen, and consultancy fees from Boehringer Ingelheim, Merck Sharp and Dohme and ViiV. Dr S Fidler has no conflicts of interest to declare. Dr Martin Fisher has received lecture fees from Abbott, Astellis, Bristol-Myers Squibb, Gilead and ViiV and he has received consultancy fees from Abbott, Bristol-Myers Squibb, Gilead, Janssen, Merck Sharp and Dohme and ViiV. Dr Andrew Freedman has received lecture and consultancy fees from Abbott, Bristol-Myers Squibb, Gilead and Tibotec/Janssen. Professor Anna Maria Geretti has received consultancy fees from Gilead and her department has received research grants from Janssen, Merck Sharp and Dohme and ViiV. Dr Yvonne Gilleece has no conflicts of interest to declare. Professor Rob Horne has no conflicts of interest to declare.

, 1963) It consists of two domains; a hydroxylase N-terminal dom

, 1963). It consists of two domains; a hydroxylase N-terminal domain with one molecule of noncovalently bound PQQ and Ca2+ at its active site as cofactors and a cytochrome c C-terminal binding domain with one covalently bound molecule of c-type haem which acts as an electron acceptor following lupanine dehydration (Hopper et al., 2002). Periplasmic targeting of the recombinant LH enzyme in Escherichia coli requires the co-expression of cytochrome c maturation factors and complex post-translational modifications that include signal peptide processing, covalent haem attachment to the C-terminal cytochrome c domain and putative disulphide bond formation

GSK126 datasheet (Stampolidis et al., 2009). Sequence analysis

with Clustal W (Larkin et al., 2007) reveals many common features of LH to members of the quinohaemoprotein family such as methanol dehydrogenase from Methylobacterium extorquens and particularly, ethanol dehydrogenase (EDH) from Comamonas testosteroni (Fig. 1). Some of the highly conserved residues among quinohaemoproteins involved in PQQ binding and at the active site of the enzyme are present click here in LH as is the invariant amino acid, Trp, which forms the floor of the active site cavity (Anthony, 1996; Hopper & Kaderbhai, 2003). In quinohaemoproteins, PQQ is usually sandwiched between a disulphide bond formed by two neighbouring Cys (Chen et al., 2002), for example, in methanol dehydrogenase 103,104Cys (Afolabi et al., 2001) and ethanol dehydrogenase 116,117Cys (Mennenga et al., 2009). The role of this bond is still a mystery. One hypothesis is that the disulphide bridge could potentially serve as an intraprotein redox centre, acting as a functional switch by relaying electrons from PQQ to the terminal acceptor in a similar manner to ferredoxin:thioredoxin reductase (Dai et al., 2000), glutathione reductase and lipoamide

dehydrogenase (White et al., 1993). A second theory claims that the bond could have a structural role for proper positioning of PQQ within the active site of the enzyme (Oubrie et al., 2002). However, LH possesses, in total, four Cys residues, two are part of the cytochrome c motif (586Cys and 589Cys), and the remaining two are separated by RVX-208 18 amino acids (124Cys and 143Cys). In this study, we attempted to establish the presence of the disulphide bond using chemical means and role in recombinant LH using site-directed mutagenesis with 143CysSer and 124,143CysSer mutations in E. coli. All chemicals were purchased from Sigma, Qiagen Ni-NTA agarose from Qiagen, and electrophoresis reagents were obtained from Bio-Rad and BDH (UK). Restriction enzymes and DNA-modifying enzymes were purchased from New England Biolabs and Promega (UK). Escherichia coli TB1 and pINK-LH-His4 construct were from Dr M. A. Kaderbhai Laboratory.

Bacterial pellets were collected by centrifugation and resuspende

Bacterial pellets were collected by centrifugation and resuspended in the purification buffer (150 mM NaCl, 20 mM Tris-HCl, pH 7.2, 0.5% Triton X-100). The samples were sonicated, and centrifuged to remove unlysed bacteria and insoluble debris. The GST-fusion proteins were purified using glutathione sepharose-4B beads according to manufacturers’ protocols (GE Healthcare) or used in other assays. For the co-purification assay, the bacterial supernatant fraction from E. coli harbouring pGEX1516/1517 was mixed with glutathione

sepharose-4B beads and agitated for 1 h at 4 °C. After washing with Tris-HCl buffer, pH 8.0, SDS-loading buffer was added and the sample was boiled for 10 min for SDS-PAGE followed by Western blot with anti-BPSS1516 antibodies. For the GST pull-down assay, a lysate from E. coli carrying pGEX-1516 expressing GST-fused BPSS1516 (GST1516) 5-FU was mixed with glutathione sepharose-4B beads. After washing off the unbound proteins, the beads with bound GST-BPSS1516 were mixed with a crude lysate from E. coli harbouring pTrc1517His and incubated for 1 h at 4 °C. The unbound proteins were removed by washing with purification buffer. The beads were analysed using SDS-PAGE and Western blotting with anti-His-tag antibodies (Abgent) and polyclonal anti-BPSS1516 antibodies. To investigate if BPSS1516 contains a secretion signal sufficient

to induce translocation of a reporter protein through the T3SS, a β-lactamase-based translocation assay was performed as described previously www.selleckchem.com/products/sd-208.html (Charpentier & Oswald, 2004). Fluorescence was measured on a Fluostar Optima Reader with excitation at 410 nm. The emission was detected via 450 nm (blue) and 520 nm (green) filters.

The measure of translocation was expressed as the emission ratio of 450/520 nm PIK3C2G to normalize the β-lactamase activity to cell loading and the number of cells present in each well. Experiments were performed in triplicate. Insertional inactivation of bpss1516 gene was performed using the suicide vector pKNOCK-1516. The plasmid was delivered into B. pseudomallei K96243 by conjugation from E. coli S17-1/λpir and transconjugants were selected on plates with 400 μg mL−1 kanamycin. The bpss1516 mutant was verified using PCR and Southern blot. Burkholderia pseudomallei invasion assays were performed according to a previously described protocol (Muangsombut et al., 2008) with the following minor modifications. An multiplicity of infection of 25 : 1 was used for an infection of 2 h. Media containing 200 μg mL−1 gentamicin plus 300 μg mL−1 spectinomycin was used for killing extracellular bacteria. To identify uncharacterized Bsa T3SS effector candidates, we analysed the published datasets of B. pseudomallei gene expression during growth in the presence of 1% arabinose (Moore et al., 2004). A locus including bpss1517 and bpss1516 was selected for further analysis because the two genes were found to be co-regulated with other Bsa-related genes (Moore et al.