H67 and H349 act as active site Zn2+ ligands in the H. influenzae DapE (Gillner et al., 2009b), with E134 shown to function as both a general acid and a general base during catalysis (Bzymek & Holz, 2004). DapE is activated by several divalent metal ions, including Zn2+, Co2+, Cd2+ and Mn2+ (Born et al., 1998; Bienvenue et al., 2003). In the presence of Mn2+, Salmonella typhi DapE functions as an aspartyl dipeptidase (Broder & Miller, 2003). DapE proteins are known to bind two divalent cations: one with high affinity (Zn2+) and the other with lower affinity (Mn2+) (Broder & Miller, 2003). DapEs exhibit a strict specificity for the l,l-isoform of SDAP (Bienvenue et al., 2003; Nocek et al., 2010). Recently, the

crystal structures of H. influenzae www.selleckchem.com/products/ABT-263.html DapE AG-014699 clinical trial with one or two zinc ions bound in the active site have been solved to 2 and 2.3 Ǻ resolution, respectively (Nocek et al., 2010). Previous to this, the 1.9 Å structure

of the apo form of DapE from N. meningitidis containing no metal ions was reported (Badger et al., 2005; Gillner et al., 2009b). Neisseria meningitidis DapE has a catalytic domain (residues 1–179 and 295–381) interrupted by a dimerization domain (180–294), and residues His68, Asp101, Glu136, Glu164 and His350 are involved in binding the two zinc atoms (Badger et al., 2005). Zn K-edge-extended X-ray absorption fine structure (EXAFS) spectra of H. influenzae DapE enzyme provided structural information on the active site and also helped establish the binding modes of phosphonate- and thiolate-containing inhibitors (Cosper et al., 2003). Two known competitive inhibitors of DapE are 2-carboxyethylphosphonic acid (CEPA) and 5-mercaptopentanoic acid (MSPA). The thiol group of MSPA binds to one or more of the Zn2+ ions in the active site of H. influenzae DapE (Cosper et al., 2003). Additionally, both l,l- and d,l-diaminopimelic acids are competitive inhibitors with respect to substrate (Born et al., 1998). A number of micromolar inhibitors of H. influenzae DapE were obtained by screening

compounds containing zinc-binding groups which included thiols, carboxylic acids, boronic acids, phosphonates and hydroxamates (Gillner et al., 2009a). The dapE deletion mutants generated in H. pylori and M. smegmatis were lethal and confirmed that dapE is essential for bacterial Oxalosuccinic acid cell growth and proliferation (Pavelka & Jacobs, 1996; Karita et al., 1997; Davis et al., 2006). The H. pylori dapE deletion mutant was unable to grow in spite of the addition of lysine to the growth medium (Karita et al., 1997; Gillner et al., 2009b). The racemization of amino acids provides meso-DAP which gets incorporated into bacterial PG (Koo & Blanchard, 1999) (Fig. 3). DAP epimerase (DapF) is a unique member of the family of pyridoxal phosphate–independent amino acid racemases, and its substrates (ll-DAP and meso-DAP) contain two stereocentres (Pillai et al., 2006).

1b). Similarly, the oligonucleotide probe gyrB2bot-DIG hybridized to the Crick ssDNA preparation, but not to the Watson ssDNA preparation, indicating minimal dsDNA contamination in the Watson ssDNA preparations

(Fig. 1c). Analysis of these Southern blot signals shows that the ssDNA preparations contained at most a ca. 10 000 : 1 ratio of ssDNA to contaminating dsDNA (Fig. 1). However, the supposedly double-stranded RF DNA preparations that were extracted from cells showed considerable ssDNA contamination (data not shown), and thus equal moles of each corresponding plasmid DNA were used for dsDNA controls in transformation Selleckchem Ribociclib experiments. Transformation with equal molar amounts of gyrB1 ssDNA was less efficient in all cases except for Crick DUS12 in MS11 than the identical dsDNA for both strains FA1090 and MS11 (Fig. 2, P < 0.05 by Student's t-test). In FA1090, Watson and Crick

DUS0 ssDNA transformation was reduced approximately 740-fold and 2200-fold, respectively, compared to matched DUS0 dsDNA (Fig. 2a, P < 0.05 by Student's t-test). Similar to DUS0 dsDNA transformation levels, DUS0 ssDNA Cabozantinib solubility dmso transformation was less efficient in MS11 than in FA1090 (Fig. 2). Interestingly, Crick DUS0 ssDNA transformation was consistently but not statistically more efficient than Watson DUS0 in ssDNA transformation (P > 0.05, threefold and twofold higher in FA1090 and MS11, respectively). In agreement with previous

reports, dsDNA transformation was enhanced by the DUS12 in both FA1090 and MS11, 6- and 16-fold compared to the DUS0 controls, respectively (Fig. 2, P < 0.05 by Student's t-test). Similarly, the Crick DUS12 sequence enhanced transformation of ssDNA in both FA1090 and MS11; however, the magnitude of enhancement was much larger than for dsDNA. The Crick DUS12 enhanced ssDNA transformation 182-fold and 467-fold over DUS0 ssDNA in FA1090 and MS11, respectively (Fig. 2, P < 0.05 by Student's t-test). In FA1090, Crick DUS12 ssDNA transformation efficiency was 24-fold lower than dsDNA DUS12 efficiency (P < 0.05 by Student's t-test). However, in MS11, Crick DUS12 ssDNA transformation efficiency was similar to dsDNA DUS12 (twofold lower, P > 0.05), which is consistent with previous findings (Stein, 1991). In contrast, Phosphoribosylglycinamide formyltransferase the Watson DUS12 ssDNA only showed a ca. sevenfold increase in transformation enhancement over matched DUS0 ssDNA (Fig. 2, P < 0.05 in FA1090, not statistically significant in MS11) and were greatly reduced from dsDNA DUS12 levels (P < 0.05, 1871-fold lower and 354-fold lower in FA1090 and MS11, respectively). The results demonstrate within ssDNA substrates that the Crick DUS12 sequence shows a much greater activity to promote transformation. Using highly purified ssDNA, we examined the ability of the Watson DUS12 or Crick DUS12 to enhance ssDNA transformation of N. gonorrhoeae.

, 2007), we rationalized that a ΔregR GSK2118436 strain might exhibit an aminoglycoside susceptibility profile similar to the bdeAB knockout strain. Our data showed, indeed, that the ΔregR mutant was at least as sensitive to kanamycin and gentamicin as the ΔbdeAB strain (Fig. 2). No significant difference was observed between the wild-type and the mutant strains when they were tested for their sensitivity against additional selected antibiotics from different classes, flavonoids, heavy metals, and detergents, among others. For a complete list of tested compounds, see Table S1. The identification of genes

for a functional MDR pump, which are coregulated with symbiotically relevant genes by RegR (Lindemann et al., 2007), raised the attractive hypothesis that BdeAB might be involved in the formation of an effective symbiosis of B. japonicum with its host plants. Soybean plants infected with the ΔbdeAB strain perhaps had a marginally increased number of nodules compared with plants infected by the wild type, but the nodule dry weight was within the wild-type range (Table 1). This shows that the mutant is not affected in its ability to nodulate. However, symbiotic nitrogen-fixation activity of the mutant was strongly decreased (Table 1), which was further manifested Selleckchem MDV3100 by the pale green-to-yellowish color of soybean leaves, a typical sign of nitrogen starvation (not shown). The symbiotic defect of the mutant was maintained

after prolonged plant growth for up to 5 weeks, which speaks against a delayed phenotype. Chromosomal integration of wild-type bdeAB genes into the ΔbdeAB mutant almost restored a wild-type level of nitrogen-fixation activity (Table 1). Confocal microscopy imaging of 3-week-old nodules elicited by the ΔbdeAB strain revealed that, while infected plant cells were densely packed with bacteroids, there was larger number of uninfected cells as compared with

nodules infected by the wild type next (not shown). To follow up on this observation, bacteroids were reisolated from 3-week-old nodules, with the result that, on average, a 10-fold lower number of viable cells were recovered from nodules infected by the ΔbdeAB strain as compared with nodules infected by the wild type (Fig. 3). The ΔbdeAB strain was also tested for its symbiotic properties on other B. japonicum host plants such as cowpea, mungbean, and siratro. Surprisingly, in contrast to soybean, the nitrogen-fixation activity of the ΔbdeAB strain was not decreased on cowpea and mungbean, and was only marginally lower on siratro, as compared with the wild type (Fig. 4). It was shown previously that the ΔregR mutant had a strong symbiotic defect on soybean (Bauer et al., 1998); however, other host plants had never been tested. While the strong symbiotic defect on soybean was confirmed, the nitrogen-fixation activity of the ΔregR mutant was far less affected on the other three hosts (Fig. 4).

This ability means that the spectrum of diseases caused by C. albicans and other Candida spp. exceeds that of most other commensal microorganisms (Calderone & Fonzi, 2001). The time-kill kinetics revealed that administration of papiliocin to C. albicans resulted in the time-dependent fungicidal rather than fungistatic activity, as was also seen after treatment with melittin

(Fig. 1). Although the killing potency of papiliocin was lower than that of melittin, this result demonstrated that the antifungal activity of papiliocin was due to the highly efficient killing of C. albicans cells. Several pathways regarding the antimicrobial mechanism of SD-208 AMPs have been proposed, including inhibition of the synthesis of specific membrane proteins, synthesis of stress proteins, arrest of DNA synthesis, breakage of single-strand DNA, interaction with DNA (without arrest of synthesis) or production of hydrogen peroxide (Andreu & Rivas, 1998). However, studies on both live SGI-1776 organisms and model membranes have indicated that most AMPs induce an increase in plasma membrane permeability. A direct correlation between the antibiotic effect and permeabilizing ability has been

found for several established AMPs such as defensins, magainins, cecropins, bactenecins or dermaseptins (Andreu & Rivas, 1998). Therefore, to investigate the mechanism of papiliocin activity, the effect of the peptide on the integrity of fungal membranes was investigated by monitoring PI influx. PI binds to DNA by intercalating between the bases with little or no sequence preference and with a stoichiometry of one dye molecule per four to five base pairs of DNA (Suzuki et al., 1997). It only enters membrane-compromised cells, after which

its fluorescence is enhanced 20–30-fold due to DNA binding Cyclooxygenase (COX) (Pina-Vaz et al., 2001; Park & Lee, 2009). If cell membranes were disrupted by papiliocin, PI could permeate into the cytoplasm and bind to fungal DNA. PI can also be used to detect pore formation (Spyr et al., 1995). The result showed that papiliocin caused an influx of PI into the fungal cells (Fig. 2). Even though the degree of influx was less potent than the influx induced by melittin, this result indicated that papiliocin could generate pores, and thereby increase the permeability of the fungal plasma membrane. Liposomes are vesicle-like structures basically constituted of phospholipids organized as concentrical bilayers containing an aqueous compartment in their interior (Cevce, 1993). Because of their amphipathic characteristics, they can incorporate substances into the aqueous compartment, the lipidic bilayer, or even distributed in both compartments (Oliveira et al., 2005). Liposomes are also used as useful tools in the construction of membrane environments. In this study, dye-entrapping liposomes were used to investigate the membrane-disruptive activity of papiliocin.

Here, we investigated the effects of the neurotransmitter serotonin and antidepressant fluoxetine (a selective serotonin reuptake inhibitor) on the modulation of adaptation-induced orientation plasticity. We show that serotonin and fluoxetine promote mostly attractive shifts. Attractive shifts augmented in magnitude towards adapter, whereas repulsive neurons reversed their

behavior in the direction of the forced orientation. Furthermore, neurons which retained their original preferred orientation expressed plasticity by shifting their tuning Cell Cycle inhibitor curves after drug administration mostly towards adapter. Our data suggest a pre-eminent role of fluoxetine by inducing and facilitating short-term plasticity in V1. “
“The suprachiasmatic nucleus (SCN) is the principal pacemaker driving circadian rhythms of physiology and behaviour. Neurons within the SCN express both classical and neuropeptide transmitters which selleck chemical regulate clock functions. Cholecyctokinin (CCK) is a potent neurotransmitter expressed in neurons of the mammalian SCN, but its role in circadian timing is

not known. In the present study, CCK was demonstrated in a distinct population of neurons located in the shell region of the SCN and in a few cells in the core region. The CCK neurons did not express vasopressin or vasoactive intestinal peptide. However, CCK-containing processes

make synaptic contacts with both groups of neurons and some CCK cell bodies were innervated by VIPergic neurons. The CCK neurons received no direct input from the three major pathways to the SCN, and the CCK neurons were not light-responsive as evaluated by induction of cFOS, and did not express the core clock protein PER1. Accordingly, CCK-deficient mice showed normal entrainment MRIP and had similar τ, light-induced phase shift and negative masking behaviour as wild-type animals. In conclusion, CCK signalling seems not to be involved directly in light-induced resetting of the clock or in regulating core clock function. The expression of CCK in a subpopulation of neurons, which do not belonging to either the VIP or AVP cells but which have synaptic contacts to both cell types and reverse innervation of CCK neurons from VIP neurons, suggests that the CCK neurons may act in non-photic regulation within the clock and/or, via CCK projections, mediate clock information to hypothalamic nuclei. “
“Ernest Gallo Clinic and Research Center at UCSF, Suite 200, Emeryville, CA, USA Intense fearful behavior and/or intense appetitive eating behavior can be generated by localized amino acid inhibitions along a rostrocaudal anatomical gradient within medial shell of nucleus accumbens of the rat.

The purposive sampling

of more non-White participants was employed, since the inclusion of ethnic minorities has been a limitation of previous studies to investigate the public’s views about community pharmacy. However, these initial findings are useful to form the basis for further qualitative (until saturation is reached) and quantitative research to establish the extent to which the general population of the UK are in support of patient registration and to identify barriers to its implementation in the future. 1. South Wales Cardiac Network. 2013. PARP inhibitor cancer New Choose Pharmacy Scheme [online]. http://www.wales.nhs.uk/sirplus/986/news/29092.pdf (Accessed 5/4/14). 2. Wilson H and Barber N. 2013. Review of NHS Pharmaceutical Care of Patients in the Community in Scotland [online]. http://www.scotland.gov.uk/Resource/0043/00430209.pdf (Accessed 4/4/13). E. Grey, H. Family, J. Sutton, M. Weiss University of Bath, Bath, UK This study explored community pharmacists’ (CPs), general practitioners’ (GPs) and practice nurses’(PNs) perceptions of teamwork to better understand what might improve CP integration into the primary care team Seventy-eight per cent of CPs considered themselves part of a multidisciplinary healthcare team (MDT), however nearly half of GPs and PNs did not include a

CP on their team GPs and PNs need to be made aware of the CP role and benefits they bring to care teams while CPs need to be more aware of the importance GPs and PNs place on face-to-face communication. The recent report on future models of care for pharmacy1 highlighted that, community pharmacy has not been fully integrated GSK2126458 into primary care

teams. Orotidine 5′-phosphate decarboxylase This may be because other health care professionals (HCPs) do not fully understand the role of the CP.1 Better integration of CPs with other HCPs on clinical teams is seen as important for enabling the extension of the pharmacist’s role and may improve patient care.2 This study aimed to explore CPs’, GPs’ and PNs’ perceptions of teamwork in order to better understand what might improve CP integration. A survey of CPs, GPs and PNs in southwest England. Closed- and open-ended questions were developed from a pilot study with pharmacists. Respondents were asked whether they considered themselves part of a MDT, then about their MDTs or whether they would like to be part of a MDT. Benefits and barriers to multidisciplinary work were also explored. The survey was available online or in paper format. Recruitment was through primary care research networks, professional journals and networks, Twitter and direct contact with practices/pharmacies. Data were entered into SPSS for statistical analysis; content analysis was used with free text responses. Ethical approval was granted by University. One hundred sixty-two CPs, 214 GPs and 147 PNs responded; response rates could not be calculated we did not know how many viewed study advertisements or social media.

It Anti-cancer Compound Library is proposed that prevented dispensing incidents frequently occurred during periods of high workload due to involuntary automaticity. Prevented dispensing incidents occurring after a busy period

were attributed to staff experiencing fatigue after-effects. “
“Objective  To find out what questions the public ask of pharmacists on a hospital medicines information helpline, and to assess the potential for improving individuals’ management of medicines through telephone helpline support. Methods  We analysed consecutive phone calls made by members of the public over 6 months to a hospital pharmacy medicines information helpline. Calls were coded for type of medicine, reason for phoning and any error revealed in the call. We also looked at which medicines were associated with harm and/or potential for harm had the caller not enquired about appropriate action to take. Key findings  Five hundred of the 923 consecutive calls to the helpline were from members of the public (including discharged hospital patients). Antimicrobial agents, analgesics and cardiovascular medicines accounted for approximately half of all calls. The reason for phoning was most often to ask about interactions (22%), directions for use (21%) or advice on adverse effects (15%). selleck inhibitor In a third of calls it is possible an error had occurred (including patient error and directions

missing from a dispensed item). Forty-eight per cent of calls were concerned with harm or judged to have potential for harm had professional information not been available. Four of these cases (0.8%), one of which was patient error and three of which were adverse effects reported by the caller, were categorised as Harm Index category F, defined as requiring intervention and referral. Conclusions  Our medicines information helpline appears to be a

valuable resource ID-8 for discharged patients and public and the advice given may be expected to improve safety with medicines and reduce harm. Our results reveal gaps in patient education about their medicines, some of which could be addressed by dispensing staff or the pharmacist at discharge. The data provide a baseline for measuring improvements in medicines management and will be useful in identifying patients who may benefit from follow-up call support from pharmacists. “
“We are delighted to welcome you to Aberdeen and to HSRPP 2014. The venue will be King’s College Conference Centre, University of Aberdeen, situated in Old Aberdeen, a historic area with architecture spanning the 15th to 21st centuries.  This is also the 20th anniversary for HSRPP and we hope that together we will celebrate this achievement and make this a memorable conference. The conference theme is “Pharmacy, Medicines and Public Health”.  This theme highlights two core components of pharmacy practice: medicines use, especially medicine safety, and public health.

Additional concerns regarding continuous ART are the induction of drug resistance, high costs, and treatment fatigue in patients. Structured treatment interruption (STI) strategies have therefore been explored in patients with viral replication suppressed under ART [3-6]. Overall, results were disappointing, with a significant

http://www.selleckchem.com/products/DAPT-GSI-IX.html proportion of patients showing rapid increases in viral load, declining CD4 T-cell counts, and an increased risk for disease progression [4, 7]. However, a subgroup of patients are able to suppress HIV replication for prolonged periods of time after STI [8]. A marker identifying such patients would be of great practical value and might renew interest in STI. Several studies have identified genetic factors influencing the pretreatment set-point viral load and time to progression to AIDS in untreated patients: the first locus identified was human leucocyte antigen (HLA)-B [9]. The natural killer (NK) cell receptor pair killer cell immunoglobulin-like receptor 3DL1 (KIR3DL1)/KIR3DS1 followed [10]. More recently, whole-genome association studies provided the information that two single nucleotide polymorphisms, found in or close to the HLA complex, both correlate with HIV viral load in untreated individuals [11]. The

first (rs9264942) is located 35 kbp upstream of HLA-C (HLA-C −35 C/T) and governs the level of surface expression of HLA-C [12]. The second (rs2395029) INK 128 lies in the HLA complex P5 (HCP5) and is in strong linkage disequilibrium with HLA B*5701, the HLA-B allele associated with the strongest protection from disease progression. Interestingly, all of

these polymorphisms are potentially associated with the function of NK cells, a subgroup of lymphocytes important in defence against viral infection: KIR3DL1 is an inhibitory receptor binding HLA-B antigens that carry the Bw4 epitope [13]. HLA-C is the ligand for the NK cell receptors KIR2DL1, KIR2DL2, KIR2DL3 and KIR2DS1 [14]. KIR–HLA interactions are important during NK cell development, as only NK cells carrying inhibitory KIR and their HLA ligands acquire full functional competence [15]. As antiviral effects of NK cells have been shown to operate most effectively in states of low viral load [16], we hypothesized that these polymorphisms may have a role in predicting Rebamipide which patients are able to maintain suppressed viral load after STI. We therefore studied the association of these polymorphisms with the evolution of viral load after STI in 130 Swiss HIV Cohort Study patients. Patients were recruited from Swiss HIV Cohort Study participants of the Strategies for Management of Anti-Retroviral Therapy (SMART), Swiss Spanish Treatment Interruption Trial (SSITT)/2nd SSITT, and STACCATO (A Trial of CD4 Guided Treatment Interruption, Compared to Continuous Treatment, for HIV Infection) trials [3-6].

, Chicago, IL, USA). The data were analyzed with descriptive statistics and chi-square test. Overall, 2,560 questionnaires were collected, 65 were incomplete and not included in our study. So, 2,495 (97.5%) questionnaires were included; the GSK458 five international airports each contributed between 391 and 629 questionnaires. The travelers had destinations in 80 countries, including 39 malaria endemic countries. All respondents were Chinese nationals with a male/female ratio of 1.55:1, of whom 2,274 (91.1%) could

access the Internet without difficulty (Table 1). Among 2,495 respondents, 1,036 (41.5%) were on their first trip and 1,459 (58.5%) had previously been abroad. The purposes of travel were tourism/holiday for 48.7%, business/work abroad for 24.9%, visit to family/friends for 10.6%, research/education for

9.8%, missionary/religious/volunteer accounted for 1.3%, and other for 4.7%. Most travelers were accompanied by a partner, their spouse, friends, colleagues, children, or other team members, while 26.7% traveled alone. While 2,069 (82.9%) travelers declared that they would stay in cities, 121 travelers (4.8%) would travel in rural areas. Among the 527 (21.1%) who intended to backpack, 285 (54.1%) were on their Tacrolimus solubility dmso first trip. High and low malaria risk destinations were visited by 1,573 (63.0%) travelers, risk-free countries by 922 (37.0%) travelers. Table 2 describes duration of stay in various risk areas. In the malaria risk group, 833 (53.0%) travelers spent less than 1 week to prepare their trip, 395 (25.1%) spent 1–2 weeks, Beta adrenergic receptor kinase 196 (12.5%) spent between 2 weeks to 1 month, 65 (4.1%) spent between 1–2 months, and 84 (5.3%) spent longer than 2 months; in the control group the numbers and proportions were 415 (45.0%), 189 (20.5%), 163 (17.7%), 58 (6.3%), and 97 (10.5%), respectively. Thus, travelers going to malaria-free destinations spent significantly more time in planning their travel (χ2 = 50.619, p < 0.001). However, among

the 527 backpackers, the preparation period was 64 and 169 days, for the risk-free and at risk countries, respectively. Among all 2,495 respondents, 1,951 (78.2%) tried to get travel health information before departure. The most common resources were the Internet (32.5%), travel agencies (27.6%), and families/friends (25.6%). Overall, 998 (40.0%) sought a travel health consultation, 65.1% and 21.0% of them did so for 1–7 days and 8–14 days before departure, respectively. The reasons why other travelers did not consult travel health professionals are listed in Table 3. There was no significant difference between the malaria risk and risk-free groups. Travelers to malaria endemic areas learned details about the infection from different sources, the main ones being family and friends (114; 7.2%) and the Internet (105; 6.7%). Only 63 (4.0%) travelers received their knowledge from travel health providers, and 181 (11.5%) received information from other medical providers. However, 905 (57.

Statistical significance of these data was analyzed using anova with multiple comparisons. Escherichia coli cells were grown in MM9 as described earlier. At an OD600 nm of 0.6–0.8, 0.2% l-arabinose was added to the cells, and when the OD600 nm reached 0.9–10, 5 μM AgNO3 was added. Cell aliquots were taken 0.25, 2.25, and 4.25 h post silver stress, were Staurosporine in vivo normalized to 3 × 108 cells mL−1, and were frozen. RNA was extracted by resuspending 3 × 108 cells in 300 μL TRIZOL reagent, phase separated using chloroform, and total RNA was precipitated using isopropanol followed by centrifugation. The RNA pellet was resuspended in nuclease-free water

(Bioexpress). Quality and purity of RNA preparations were assessed by electrophoresis and via spectrophotometric determination of the ratio of absorbance at 260/280 nm. Total-RNA extracted from the previous step was treated with RNase-free DNaseI (Fermentas). First-strand cDNA was prepared from 2 μg of total RNA using the Superscript III cDNA synthesis kit (Quanta Biosciences). The cDNA was then diluted with SYBR green qPCR master mix. Reactions HIF inhibitor were amplified using the Applied Biosystems 7300 Real-time PCR system. Each cDNA sample was assessed in triplicate using 16S-rRNA gene as an internal control. Thermal cycle conditions consisted

of an initial denaturation step at 95 °C for 60 s followed by 40 cycles of 95 °C for 15 s and 60 °C for 60 s. Fluorescence was measured at the beginning of each annealing/extension step. Amplicon size was also determined using electrophoresis on an agarose gel (1% w/v). The quantity of cDNA measured by real-time PCR was normalized to the abundance

of 16S cDNA. Primers used for qRT-PCR are listed in Table S1. To check the specificity of each primer, the predicted amplicon melting temperature was confirmed via dissociation curve analysis. Relative expression from the cusC gene was calculated using the ΔΔCt quantification method (Livak & Schmittgen, 2001), and values are averages of three independent experiments. Statistical significance of these data was analyzed using anova with multiple comparisons. The role of copper ions in bacterial growth Protein tyrosine phosphatase and survival is well documented. Owing to the toxic nature of copper ions, bacteria such as E. coli and Salmonella have molecular systems that tightly control the copper concentration within the cells (Pontel & Soncini, 2009). In E. coli, the Cus system was first identified as a silver resistance system and was shown to be inducible by copper ions as well. Upon further investigation, it was revealed that the chemiosmotic CusCFBA system in E. coli confers anaerobic copper and silver resistance and is regulated by the CusR/CusS two-component system. The sensor kinase CusS and the response regulator CusR are activated by copper (Munson et al., 2000) and silver (Franke et al., 2001) ions, and these proteins are required for regulation of the cusCFBA operon. To define the role of CusS in copper resistance in E.