LB plates were supplemented with ampicillin (100 μg mL−1), or kanamycin (35 μg mL−1) when necessary. Both M9 liquid media and agar plates supplemented with 0.4% glucose were used for evaluating the superoxide resistance phenotype. The indicator 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-Gal) was added to LB plates at a final

concentration of 40 μg mL−1. Cells were treated with 50 μM PQ, 5 mM SAL, or 5 mM DIP and were incubated at 37 °C with shaking for 1 h where indicated. Susceptibility testing was performed on Mueller Hinton (MH) plates. mdtB::frt, mdtF::frt, acrB::frt, emrB::frt, acrD::frt, macB::frt, mdtC::frt, acrE::frt, Palbociclib supplier emrY::frt J. L. Rosner and R. G. Martin, in preparation The microarray analysis procedure was performed as previously reported (Fabrega et al., 2010). Briefly, 20 μg of total RNA extracted from a mid-exponential phase culture was labeled with Cy-3-dUTP (RNA from strain PS5) or Cy-5-dUTP (RNA from strain NorE5). Labeled cDNA samples were hybridized for 5 h at 65 °C with the DNA microarray chip, which contained 4058 open reading frames (ORFs) LY2109761 representing 95% of E. coli ORFs. Axon Scanner GENPIX 1.0 was used to obtain the resulting 16-bit TIFF images that were analyzed using scanalyze software (http://rana.stanford.edu/software/). The reproducibility of the technique

was assessed in three separate experiments. A normalized relative Cy5/Cy3 ratio > 2 was considered as a significant increase in expression and a normalized relative Cy3/Cy5 ratio > 2 was considered as a significant decrease in expression when observed for all the three different experiments performed. RNA extraction and analysis was performed according to a previous study (Fabrega et al., www.selleck.co.jp/products/Metformin-hydrochloride(Glucophage).html 2009). Briefly, strains were incubated until they reached OD600 nm values of 0.5–0.6. Cultures were

mixed with RNA protect Bacteria Reagent (Qiagen) and subsequently treated with TE buffer supplemented with lysozyme. Total RNA was extracted using RNeasy Mini Kit (Qiagen), and samples were then treated with DNA-free DNase (Ambion). RT-PCR was performed using the AccessQuick RT-PCR System (Promega). gapA (a housekeeping gene) was defined as the internal control. The retrotranscription process was performed using 500 ng of RNA at 45 °C for 45 min followed by a standard PCR program. Results were corroborated from two independent RNA extractions and amplifications. A lacZ transcriptional fusion was constructed with the ompN80 and the ydbK49 fragments (Simons et al., 1987). Amplification of both fragments was carried out by using strain GC4468 as template. The 405 bp ompN80 fragment started from −384 to +21, relative to the ATG, whereas the 427 bp ydbK49 fragment started from −401 to +26 (Fig. 1). The amplified DNA fragments were digested with EcoRI and BamHI and ligated to the similarly cut vector pRS551. Recombinant plasmids were isolated in DH5α cells by selection for ampicillin resistance and verified by sequence analysis.

Given this developmental shift, the AVMMR may represent a less mature electrophysiological pattern of AV speech processing because it was associated with less time spent looking at the articulatory movements during speech. The maturational changes in the way auditory and visual information is processed by younger and older infants are reflected in developmentally transient ERP components, which are reliably elicited in younger infants but are not always observable in older infants and/or adults. For instance, the AVMMR recorded in 2-month-old infants by Bristow et al. (2009) was not observed in adults (G. Dehaene-Lambertz,

EPZ015666 cost personal communication; see also Jääskeläinen et al., 2004), and an increase in the visual N290 component to static direct eye-gaze vs. averted eye-gaze reported in 4-month-old infants (Farroni et al., 2002) was not observed in 9-month-old JAK inhibitor infants (Elsabbagh et al., 2009) or adults (Grice et al., 2005). In order to further explore the question of the developmental profile of the AVMMR neural response, a group of adults was also tested (see Control study S3 and Fig. S7). No AVMMR in response to either audiovisually incongruent (combination and fusion) stimuli was observed, confirming our hypothesis that this component indicates a less mature type of processing of AV conflict only in early infancy. [Note that the present study did

not employ an oddball paradigm used in previous adult studies (Saint-Amour et al., 2007; Hessler et al., 2013), where AVMMR was elicited in response to the deviant among repetitive standards and not to the AV violation per se. Therefore,

the absence of the AVMMR in the present study does not contradict the results of the above studies but, on the contrary, provides corroborative evidence that adults perceived the two incongruent conditions integrated.] It is not surprising therefore that while the AVMMR was observed at the group level in younger infants (4.5–5.5 months, N-acetylglucosamine-1-phosphate transferase Kushnerenko et al., 2008; and 2-month-old, Bristow et al., 2009), it was only found in the present study in a subset of our infants, who demonstrated a less mature pattern of looking behaviour. It is important to note here that the group-averaged ERP results might obscure the meaningful individual differences in the level of maturation of multisensory processing in individual infants. Thus, it appears that the AVMMR is a developmentally transient ERP response that may begin to disappear around the age of 6–9 months, similar to mismatch positivity (or PC) in young infants (Morr et al., 2002; Kushnerenko, E., Van den Bergh, B.R.H., & Winkler, I. (under review)). The developmental decrease in the auditory PC during the first year of life was suggested to reflect decreasing sensitivity to less informative sensory cues, which was initially high in younger infants (Kushnerenko, E., Van den Bergh, B.R.H., & Winkler, I. (under review)).

Given this developmental shift, the AVMMR may represent a less mature electrophysiological pattern of AV speech processing because it was associated with less time spent looking at the articulatory movements during speech. The maturational changes in the way auditory and visual information is processed by younger and older infants are reflected in developmentally transient ERP components, which are reliably elicited in younger infants but are not always observable in older infants and/or adults. For instance, the AVMMR recorded in 2-month-old infants by Bristow et al. (2009) was not observed in adults (G. Dehaene-Lambertz,

p38 MAPK pathway personal communication; see also Jääskeläinen et al., 2004), and an increase in the visual N290 component to static direct eye-gaze vs. averted eye-gaze reported in 4-month-old infants (Farroni et al., 2002) was not observed in 9-month-old learn more infants (Elsabbagh et al., 2009) or adults (Grice et al., 2005). In order to further explore the question of the developmental profile of the AVMMR neural response, a group of adults was also tested (see Control study S3 and Fig. S7). No AVMMR in response to either audiovisually incongruent (combination and fusion) stimuli was observed, confirming our hypothesis that this component indicates a less mature type of processing of AV conflict only in early infancy. [Note that the present study did

not employ an oddball paradigm used in previous adult studies (Saint-Amour et al., 2007; Hessler et al., 2013), where AVMMR was elicited in response to the deviant among repetitive standards and not to the AV violation per se. Therefore,

the absence of the AVMMR in the present study does not contradict the results of the above studies but, on the contrary, provides corroborative evidence that adults perceived the two incongruent conditions integrated.] It is not surprising therefore that while the AVMMR was observed at the group level in younger infants (4.5–5.5 months, Selleck Paclitaxel Kushnerenko et al., 2008; and 2-month-old, Bristow et al., 2009), it was only found in the present study in a subset of our infants, who demonstrated a less mature pattern of looking behaviour. It is important to note here that the group-averaged ERP results might obscure the meaningful individual differences in the level of maturation of multisensory processing in individual infants. Thus, it appears that the AVMMR is a developmentally transient ERP response that may begin to disappear around the age of 6–9 months, similar to mismatch positivity (or PC) in young infants (Morr et al., 2002; Kushnerenko, E., Van den Bergh, B.R.H., & Winkler, I. (under review)). The developmental decrease in the auditory PC during the first year of life was suggested to reflect decreasing sensitivity to less informative sensory cues, which was initially high in younger infants (Kushnerenko, E., Van den Bergh, B.R.H., & Winkler, I. (under review)).

The DSR system presumably forms intracellular sulfite that is oxidized by an enzyme system consisting of Sat, Apr, and Qmo proteins (Rodriguez et al., 2011). The electron acceptors, cytochrome c, and menaquinone (Fig. 1) are ultimately oxidized

by the photosynthetic reaction center. In cultures of Cba. tepidum that contain both sulfide and thiosulfate, sulfide is oxidized preferentially while sulfur globules are formed (Chan et al., 2008; Azai et al., 2009; Holkenbrink et al., 2011). Following sulfide depletion, thiosulfate and sulfur globules are oxidized to sulfate. The molecular mechanism of Alpelisib mw this phenomenon is poorly understood. Sulfide possibly inhibits thiosulfate oxidation either by substrate competition between sulfide and thiosulfate (the SOX system oxidizes sulfide in vitro; Ogawa et al., 2010) or by saturation of the electron acceptor pool. Regulation of sulfur metabolism genes in GSB is poorly described, but it is known that SoxA is induced by thiosulfate in Chlorobaculum thiosulfatiphilum (Verté et al. 2002). find more In the purple sulfur bacterium, Allochromatium vinosum, sox and dsr genes are expressed at a low constitutive level in the absence of reduced sulfur substrates and are induced by thiosulfate and sulfide, respectively (Grimm et al., 2010, 2011). Chlorobaculum tepidum TLS was grown under incandescent illumination in CL medium (Frigaard et al., 2004). For experiments comparing early and late exponential growth phase, wild-type

cells were grown at 45 °C in 1-L flasks under a light intensity of 200 μmol photons m−2 s−1. For experiments comparing wild type and the dsrM mutant (Holkenbrink et al., 2011), cells were grown at 42 °C in 15-mL

tubes under a light intensity of 50 μmol photons m−2 s−1 and harvested in the late exponential growth phase. Cells were harvested by centrifugation and stored at −20 °C prior to analysis. Bacteriochlorophyll c was determined by extracting the cell pellet with acetone : methanol (7 : 2 by vol) (Frigaard et al., 1997). Sulfide was measured using the colorimetric methylene blue method (Cline 1969). Sulfate Ribonucleotide reductase and thiosulfate were measured by ion chromatography (Dionex, Hvidovre, Denmark) using a carbonate buffer as eluent. Samples for analysis of elemental sulfur were dissolved in methanol and analyzed as S8 by HPLC using a Sykam pump (S1100), UV–VIS detector (Sykam S3200), Zorbax ODS-column (125 × 4 mm, 5 μm; Knauer, Berlin, Germany) and methanol as the eluent at a flow rate of 1 mL min−1. Elemental sulfur was detected at 265 nm. Cell pellets were thawed and extracted in acetone : methanol (7 : 2 by vol) to remove pigments. The colorless cell pellets were solubilized in an SDS-containing buffer (Laemmli, 1970) supplemented with a complete protease inhibitor cocktail (Roche, Hvidovre, Denmark) at 95 °C for 3–5 min and cleared by centrifugation. Prior to digestion, proteins were reduced with dithiothreitol (1 mM) and alkylated with iodoacetamide (5 mM).

, 2009; Toledo-Arana et al., 2009). An alternative use of high-throughput sequencing has been in the sequencing of immunoprecipitated RNA or DNA (IP-seq), which is an alternative to ChIP-on-chip experiments (Wade et al., 2007). A recent example of such an approach has

been the simultaneous identification of sRNA and mRNA of S. enterica serovar Typhimurium, which were bound to the RNA chaperone Hfq (Sittka et al., 2008). The rapid developments in sequencing technologies allow one to obtain very Ku0059436 high-definition transcription snapshots, and these will, undoubtedly, significantly increase our insights in transcriptional and post-transcriptional events in microorganisms. Besides the increased insight into the process of transcription, it will also help in improving or correcting the annotation of

genome sequences (Denoeud et al., 2008). Identification of the 5′ and 3′ boundaries of mRNA species will inform us of the most likely translation initiation codon, especially in those cases where a ribosome-binding site is not apparent (Moll et al., 2002). Next to technical challenges, the rapid increases in knowledge Venetoclax in vivo will be accompanied by new problems, as with previous breakthroughs in functional genomics (like genome sequencing and microarrays). Several issues may require action from the scientific community, and some of these are highlighted below. 1 Differentiation of transcriptional

and post-transcriptional events. The sequencing-based approaches used for determining the bacterial transcriptomics to date are not able to distinguish between de novo transcription and post-transcriptional events, as they only record the levels of RNA (cDNA) present. This is a weakness shared with microarray technology. Alternative approaches such as those used for genome-wide determination of transcription start sites by 5′ rapid amplification of cDNA ends (RACE) and 5′-serial analysis of gene expression approaches (Hashimoto BCKDHA et al., 2004, 2009). These approaches use techniques distinguishing between primary (capped) RNA species, which result from de novo transcription, and processed (uncapped) RNA species. The combination with standard RNA-seq allows for specific identification of primary transcripts, and could be coupled to the use of rifampicin to inhibit transcription for the study of RNA stability (Mosteller & Yanofsky, 1970). Historically, research on microbial transcription focused on protein-based signal transduction and regulatory systems, and mRNA was seen as a relatively inert information carrier. However, the conventional view of RNA has changed in the last decade due to the discovery of regulatory and catalytic RNA activity (Waters & Storz, 2009).

For all experimental assays, 24-well tissue culture plates (Greiner) were seeded with 5.0 × 104 HEp-2 cells. Plates were incubated for 18 h at 37 °C in a humidified 5% CO2 incubator. Before the assays, the semi-confluent monolayers were washed and incubated with RPMI Medium 1640 containing 1% fetal bovine serum. Planktonic and biofilm SS cells were suspended in fresh medium to a final concentration of 107 cells mL1. Bacterial CFU were confirmed by plating onto THB agar. Cells were aliquoted

into 24-well tissue culture plates (1 mL per well). All 24-well plates were incubated under a 5% CO2 atmosphere at 37 °C for 3 h to allow cells to attach. After 3 h of incubation, all plates were washed three times with PBS to remove any nonadhering bacteria. Adherent cells were detached using 0.25% trypsin, serially diluted 10-fold in sterile PBS and plated onto THB agar plates. Results are expressed as the Sirolimus average number of bacteria adhering to HEp-2 cells for determinations. Negative control wells containing only HEp-2 cells were used in all experiments. The assay was performed at least three times. Planktonic cultures were grown in THB medium with 1% fibrinogen at 37 °C Torin 1 datasheet and were collected after 24 h culture time. For biofilm cultures, SS2 HA9801 was grown

in a 100-mm tissue culture dish (Greiner) at 37 °C for 24 h. Planktonic and biofilm cells were harvested and total cellular RNAs were extracted as described previously with little modification (Shemesh et al., 2007). Total RNA was extracted with an E.Z.N.A.™ Bacterial RNA isolating kit (Omega, Beijing, China) according to the manufacturer’s protocol.

The RNA was subjected to DNase I (Promega, Madison, WI) treatment to exclude genomic DNA contaminants. cDNA synthesis was performed using the PrimeScript™ RT reagent kit (TaKaRa) according to the manufacturer’s instructions. mRNA levels were measured using two-step relative qRT-PCR. Relative copy numbers and expression ratios of selected genes were normalized to the Cytidine deaminase expression of one housekeeping gene (16S rRNA gene) and calculated as described by Gavrilin et al. (2000). The housekeeping gene (16S rRNA gene) was used as an internal control for specific primers. The specific primers used for the various RT-PCR assays are listed in Table 1. The SYBR Green PCR method was used according to the SYBR Premix Ex Taq™ Kit (TaKaRa). Reactions were carried out in triplicate. An ABI 7300 RT-PCR system was used for relative qRT-PCR. Planktonic and biofilm cells were inactivated for 1 h at 90 °C (Azad et al., 1999). Efficiency of inactivation was determined by plating the above bacterial suspension onto THB and the presence of bacterial colonies was monitored for 3 days. Adult zebrafish were divided randomly into three groups (100 fish per group).

“
“Recently, travel to underdeveloped and exotic destinations has increased substantially. International travel is a multi-billion dollar industry exceeding $900 billion US Navitoclax dollars (600 euros) in 2008. By the year 2020, it is expected

that the number of international travelers will exceed 1 billion, half being for leisure purposes and approximately 15% business related.1Prior to departure for travel, it is widely recommended to consult with a specialist in travel health, as many travelers are unaware of the immunizations and preventative measures that are recommended. Pharmacists are accessible healthcare professionals who have unique opportunities to provide education and administration of immunizations learn more to this population. Over the past two decades, pharmacists have become more involved in the provision of travel medicine services in a variety of settings.2–5 The Clinical Pharmacy

International Travel Clinic (CPITC), established in the early 1990s, is a telepharmacy consultation service run by pharmacists from the Kaiser Permanente Colorado region.2 The team, composed of five clinical pharmacists, a pharmacy technician, and a consulting infectious diseases physician, provide phone consultation for approximately 9500 travel patients every year, following referrals from primary care physicians (PCPs) or customer service associates. As no appointments are required, patients receive their consultation at the time they call the service. The pharmacists provide recommendations regarding travel immunizations, medications, and preventive measures against diseases abroad, and they attain prescriber co-signatures for these orders. It is estimated that the CPITC pharmacists could save $47,000 per year in unnecessary immunizations with this consultation service.3 Community pharmacists have also become involved in travel medicine services, due to their ease of accessibility with many convenient locations, long hours of operation, and the ability to immunize.3,5 One pretravel health program, TravelRx, offered by a supermarket

chain pharmacy in Central Virginia, provides initial phone consultation followed by individualized appointments in a private counseling room within the pharmacy for approximately 1000 patients per year.4Following the patient interview and assessment of travel-related needs, the patient’s Neratinib mw PCP is contacted to gain authorization for the administration of immunizations and medications; the pharmacist then schedules an appointment for the patient’s travel education and immunizations. Following the patient’s visit, the pharmacist follows up with both the physician (to provide documentation of the patient’s immunizations) and the patient (to complete any additional vaccine series post-travel). Patients expressed a high level of satisfaction with the pharmacist-run program through patient satisfaction surveys, although no outcomes were formally assessed.

Multiple mechanisms might be responsible for generating the observed

diversity in 5S rRNA genes in a genome. In organisms containing multiple rRNA genes, the homogeneity of primary structures is believed to be maintained through gene conversion by homologous recombination (Hashimoto et al., 2003), as a form of concerted evolution (Abdulkarim & Hughes, 1996). Although the observed homogeneity of 5S rRNA genes in the majority of species analyzed could be attributed to the effect of homologous recombination, the recombination appeared to be compartmentalized or ineffective in some genomes. The observed high degree of diversity in the primary structures of the 5S rRNA genes in partial or split rRNA gene operons and the rrnC operon in T. tengcongensis suggested that these rRNA genes have been excluded www.selleckchem.com/products/Roscovitine.html from participation in concerted evolution. Such compartmentalization was also present in B. subtilis that has two similarity groups of rRNA genes appeared to have evolved

independently, as evidenced by their relation to different 5S rRNA genes-rrn23S spacers. Despite the lack of sequence homogeneity, secondary structures of these genes were well conserved, most likely due to the life and death driving force of ribosomal constraints. Compared with whole 16S and 23S rRNA genes, 5S rRNA genes are a less ideal taxonomical marker for use in analyses of complex microbiomes. The main reason is the widespread intragenomic 5S rRNA gene diversity. Approximately, 12.3% (96 of 779) selleck products of the unique

species analyzed had > 3% intragenomic variation of 5S rRNA genes, compared to only about 1% of species with similar degree of variation in 16S and 23S rRNA genes (Coenye & Vandamme, 2003; Acinas et al., 2004; Pei et al., 2010). This high degree of diversity most often occurs between a standalone 5S rRNA Dehydratase gene (orphan or split) and a 5S rRNA gene in a complete rRNA operon. The lack of standalone 16S or 23 S rRNA genes appears to be the main reason for the lower intragenomic diversity among 16S or 23S rRNA genes. Orphan 5S rRNA genes are sometimes overlooked by a whole-genome annotation program because of their small size. Compared with rrnDB (Lee et al., 2009), a publically accessible database that collects existing data on structure RNA genes from whole-genome sequencing projects, 11 genomes listed in Table 1 that had additional 5S rRNA genes in our study are not listed in rrnDB. The additional 5S rRNA genes would have been invisible if blast search of 5S rRNA genes against the whole genomes were not performed. Nevertheless, in 26 of the 52 genomes listed in Table 1, correct records of the orphan 5S rRNA genes can be found in rrnDB. The remaining 15 of the 52 genomes have no entries in rrnDB. Divergent evolution between paralogous 5S rRNA genes in a genome may corrupt the record of evolutionary history and obscure the true identity of an organism.

Multiple mechanisms might be responsible for generating the observed

diversity in 5S rRNA genes in a genome. In organisms containing multiple rRNA genes, the homogeneity of primary structures is believed to be maintained through gene conversion by homologous recombination (Hashimoto et al., 2003), as a form of concerted evolution (Abdulkarim & Hughes, 1996). Although the observed homogeneity of 5S rRNA genes in the majority of species analyzed could be attributed to the effect of homologous recombination, the recombination appeared to be compartmentalized or ineffective in some genomes. The observed high degree of diversity in the primary structures of the 5S rRNA genes in partial or split rRNA gene operons and the rrnC operon in T. tengcongensis suggested that these rRNA genes have been excluded Cell Cycle inhibitor from participation in concerted evolution. Such compartmentalization was also present in B. subtilis that has two similarity groups of rRNA genes appeared to have evolved

independently, as evidenced by their relation to different 5S rRNA genes-rrn23S spacers. Despite the lack of sequence homogeneity, secondary structures of these genes were well conserved, most likely due to the life and death driving force of ribosomal constraints. Compared with whole 16S and 23S rRNA genes, 5S rRNA genes are a less ideal taxonomical marker for use in analyses of complex microbiomes. The main reason is the widespread intragenomic 5S rRNA gene diversity. Approximately, 12.3% (96 of 779) Etoposide mw of the unique

species analyzed had > 3% intragenomic variation of 5S rRNA genes, compared to only about 1% of species with similar degree of variation in 16S and 23S rRNA genes (Coenye & Vandamme, 2003; Acinas et al., 2004; Pei et al., 2010). This high degree of diversity most often occurs between a standalone 5S rRNA Vildagliptin gene (orphan or split) and a 5S rRNA gene in a complete rRNA operon. The lack of standalone 16S or 23 S rRNA genes appears to be the main reason for the lower intragenomic diversity among 16S or 23S rRNA genes. Orphan 5S rRNA genes are sometimes overlooked by a whole-genome annotation program because of their small size. Compared with rrnDB (Lee et al., 2009), a publically accessible database that collects existing data on structure RNA genes from whole-genome sequencing projects, 11 genomes listed in Table 1 that had additional 5S rRNA genes in our study are not listed in rrnDB. The additional 5S rRNA genes would have been invisible if blast search of 5S rRNA genes against the whole genomes were not performed. Nevertheless, in 26 of the 52 genomes listed in Table 1, correct records of the orphan 5S rRNA genes can be found in rrnDB. The remaining 15 of the 52 genomes have no entries in rrnDB. Divergent evolution between paralogous 5S rRNA genes in a genome may corrupt the record of evolutionary history and obscure the true identity of an organism.

NHS research ethics approval was not required. A piloted questionnaire was sent to the pharmacist in charge at a stratified random sample of 500 community pharmacies in England, Wales and Scotland, Cyclopamine with a reminder sent to non-respondents after four weeks. An online version of the questionnaire was produced using SurveyMonkey; participants were recruited via the Royal Pharmaceutical Society Great Western Local Practice Forum, LocumVoice and Pharmacy Forum discussion forums to increase the number of potential respondents. SPSS v20 was used for statistical analysis. 222 responses were returned by Freepost

(44% response). 209 responses were received via SurveyMonkey (response rate not calculated due to open nature of forums). Aqueous Cream BP would be recommended as an emollient by 43% (96/222) Freepost respondents and by 35% (73/209) online respondents (n.s.), with 24% (49/208) of the Freepost respondents and 12% (24/199) online respondents recommending it first-line (χ2 = 9.1, df = 1, p = 0.003). Recently-registered pharmacists (2009–2012) were more likely [48% (36/75)] to recommend Aqueous Cream BP than those qualifying between 2002–2008 [44% (40/92)], 1985–2001 CX5461 [39% (48/122)] or earlier [28% (31/109)] (Mantel-Haenszel linear-by-linear association χ2 = 7.8, df = 1, p = 0.005). The majority (57%) of all respondents were less likely to recommend Aqueous Cream BP as an emollient than they were three

years ago. Results showed variation between the two cohorts in the reference sources used in response to dermatology queries, with respondents who replied via SurveyMonkey more likely to use e-resources than those who responded via Freepost who were more likely to use paper-based or employer-provided information sources. Recent communication from the MHRA has again emphasized

the problems that SLS can cause, especially in children, when used in emollients.2 While a limited study with a relatively small sample, and a contrasting cohort, these results show that a significant minority of community pharmacists are still recommending Aqueous Cream BP as an emollient. It is somewhat curious that more-recently educated pharmacists are more likely to do so and the reasons for this require further investigation. Given the variations in information sources used by the two cohorts of respondents, Immune system an appropriate variety of educational interventions is required to improve practice by updating textbooks, responding to symptoms guides and e-guides. Minor ailment scheme lists were not investigated as part of this study and may also need review. 1. Tsang M, Guy RH. Effects of Aqueous Cream BP on human stratum corneum in vivo. British Journal of Dermatology 2010; 163: 954–958. 2. MHRA. Aqueous cream: may cause skin irritation, particularly in children with eczema, possibly due to sodium lauryl sulfate content. Drug Safety Update March 2013; 6: A2.