A 58.8–60% increase in ROS was caused by glutathione in the strain in which there was a significant decrease in the MIC (resistant S. aureus 22), whereas in the sensitive strain, glutathione increases the production of ROS only by 12.8–16.6%, without any significant change occurring in MIC. There was a correlation between the stimulus of ROS and the decrease of MIC caused by exogenus glutathione. The glutathione stimulated intracellular ROS, even in strains without the antibiotic, and also increased the oxidative selleck stress at all concentrations of the antibiotics assayed. However, this enhancement was more marked at the higher concentrations of both antibiotics (Figs 3 and 4). The

exogenous glutathione decreased the extracellular ROS, up to a maximum of 86% in the two strains treated with ciprofloxacin, with similar results being obtained with gentamicin. It was previously Selleck AZD6244 shown that synthetic quinolone antibiotics

promoted the formation of the hydroxyl radical that contributed to cell death (Kohanski et al., 2007), and it was proposed that oxidative damage contributes to bactericidal cell death following gyrase poisoning with an oxygen-dependent death pathway appearing to amplify the primary effect on gyrase (Dwyer et al., 2007). Glutathione was chosen because it is a scavenger of ROS, which has been shown to be involved in protecting the cell either directly or indirectly. This might constitute an adaptive response to oxidative damage, which is known to increase in the presence of the antibiotic (Prinz et al., 1997; Carmel-Harel & Storz, 2000; Pomposiello & Demple, 2002). Compounds such as glutathione can rapidly cross the cell L-NAME HCl membrane, due to their hydrophobic nature, low molecular weight and the presence of specific transporters for these antioxidants in the cell membrane, thus allowing them to produce an antioxidant action in the cytosol (Parry & Clark, 2002; Zhang et al., 2003). A previous study conducted on Escherichia coli suggests that glutathione modulates

the effect of antibiotics (Goswami & Jawali, 2007). These authors reported a reduction in MIC for ampicillin and penicillin, from 8 to 4 μg mL−1 and from 64 to 48 μg mL−1, respectively, which is not as marked as that found in our study for ciprofloxacin and gentamicin in S. aureus. According to our results, there exists the possibility of modifying the sensitivity of resistant strains of S. aureus by the addition of glutathione. These antecedents sustain the hypothesis of our work, which suggests that the antioxidants are useful to improve the bactericidal action of ciprofloxacin. Considering that the antioxidant defense in S. aureus is transcriptionally regulated, and that the expression of oxyR genes occurs in response to external conditions via a glutathione-dependent redox enzyme (Zheng et al., 2001; Uziel et al.

This is also valid for Frankia that fix nitrogen both in free-living and in symbiotic conditions. Frankia symbiosis results from interaction between the Frankia bacteria and dicotyledonous plants, that is, actinorhiza. These plants, which are important in forestry and agroforestry, form, together with the

legumes (Fabales), a single Talazoparib order nitrogen-fixing clade. It has been shown that a receptor-like kinase gene, SymRK, is necessary for nodulation in actinorhizal plants as well as in legumes and arbuscular mycorrhizal fungi. Recently, the involvement of isoflavonoids as signal molecules during nodulation of an actinorhizal plant was shown. The genome sizes of three Frankia species, Frankia EANpec, ACN14a and CcI3, are different, revealing a relationship between genome size and geographical distribution. Recent genomic sequencing data of Frankia represent genomes from cluster I to IV, indicating that the genome of DgI is one of the smallest genomes

in Frankia. In addition, nonsymbiotic Frankiales such as Acidothermus cellulolyticus, Blastococcus saxoobsidens, Geodermatophilus obscurus and Modestobacter marinus have a variety of genome sizes ranging from 2.4 to 5.57 Mb. “
“Some Candida species are common commensals, which can become Osimertinib opportunistic pathogens in susceptible hosts. In severely ill patients, Candida species, particularly Candida albicans, can cause life-threatening systemic infections. These infections are difficult to diagnose, as symptoms are similar to those of systemic bacterial infections. These difficulties can lead to delays in initiation in antifungal therapy, which contributes to the C-X-C chemokine receptor type 7 (CXCR-7) high mortality rates (>40%) associated with these infections. In order to investigate systemic Candida infection, mouse models have been developed that mimic human disease, the most common being the intravenous infection model and the gastrointestinal colonization

and dissemination model. This review discusses the two models and the contributions that they have made to our understanding of fungal virulence, host response to infection and the development of novel antifungal therapies and diagnostics. A select number of Candida species are usually found as harmless commensals in the gastrointestinal tract, oral cavity and genital area of healthy individuals. Candida spp. can be isolated from the majority of healthy individuals, with the highest fungal counts found in the duodenum (Kusne et al., 1994). The most common species isolated is Candida albicans, with Candida parapsilosis, Candida glabrata, Candida tropicalis, Candida dubliniensis and Candida krusei also found (Kusne et al., 1994; Scanlan & Marchesi, 2008).

The cultures were centrifuged at 5000 g for 20 min at 4 °C. The resultant pellet was resuspended in 3 mL of lysis buffer (Tris-HCl 50 mM, NaCl 100 mM, 50 μg mL−1 lysozyme, pH 8), and incubated at 37 °C for 30 min. The samples were sonicated at 11 r.m.s. (three pulses of 20 s)

and centrifuged at 16 000 g for 45 min at 4 °C. The protein concentration of supernatants was determined by BCA Protein Assay Kit (Thermo Corporation) according to the manufacturer’s instructions. Finally, 2 μg of P. salmonis RNA was incubated with 100 μg of the E. coli protein extract for 1.5 h at 37 °C. As a positive control, 2 μg of RNA was treated with commercial RNase A (E.Z.N.A Omega-Biotek) and as negative control 2 μg of P. salmonis RNA

alone was incubated under the same conditions described above. The digested RNA was visualized on 1% agarose gel stained with GelRed™. The GenBank accession number for the P. salmonis ps-Tox-Antox Ibrutinib locus is HQ008719. The resultant sequences were analysed by FgeneB tool, finding that a sequence of 905 bp contains two putative ORFs. The ORF1 encodes a putative protein of 75 amino acid residues and the ORF2 encodes a putative protein with 135 amino acid residues. Both amino acid sequences were submitted to blastp analysis to determine protein identities. The blastp analysis shows that the protein encoded by the ORF1 has a high level of similarity to antitoxin proteins Small molecule library of bacterial TA modules, specifically to VapB and VagC antitoxins (Table 1). The product of the ORF1, named Ps-Antox, contains an SpoVT/AbrB domain, which is a DNA-binding domain, and, as such, belongs to the super family of transcriptional regulators of the same name. The protein encoded by ORF2, named Ps-Tox, seems to be strikingly

similar to toxin proteins of bacterial TA modules, specifically the VapC toxin (Table 1). Additionally, the protein encoded by ORF2 shows the presence of a PIN domain (a homologous domain to the N-terminal domain of the pili biogenesis protein PilT), which is highly conserved in the VapC homologues. The sequence alignment of the Ps-Tox, with other homologues Nintedanib (BIBF 1120) VapC proteins of bacterial TA modules shows a high degree of conservation between them (see Supporting Information, Fig. S1). These results indicate that we have found a typical TA locus in the genome of P. salmonis, named Ps-Tox-Antox. The P. salmonis ps-Tox-Antox locus consists of a bicistronic operon conformed by an upstream 228-bp gene (ps-Antox) and a downstream 408-bp gene (ps-Tox) separated by an 8-bp intergenic spacer (Fig. 1). By analysis with bprom, we have found a putative promoter region and a Shine–Dalgarno sequence upstream of the ps-Antox gene (Fig. 1). This putative promoter contains a pair of 7-bp inverted repeat sequences (IRs) between the −10 and −35 regions, which is characteristic of other TA operons.

For this study, we selected all participants who entered the SHCS between 1 January 1996 and 31 December 2008. The seven SHCS centres, 13 affiliated hospitals and 33 private collaborating physicians from all regions of the country were addressed in formal correspondence to provide aggregated (i.e. unidentifiable) numbers of HIV-positive persons not participating in the SHCS during their first clinical visit in 2008. Collected information

included geographical region of origin, gender, injecting drug use (IDU) and whether the patient was on ART. After two rounds of reminders via email and/or telephone, the response rate was 40 of 53 (75%) clinics or private physicians, and those that responded included all seven SHCS centres and all large institutions providing HIV care. Among participants not known to have died, we defined LTFU as no further cohort visit during at least 1 year after the last visit. find more We distinguished seven geographical regions of patients’ origin Selleckchem CT99021 according to an adopted UNAIDS classification of nationalities [15]. Because of the small numbers of persons in care and their similar

demographic characteristics, we merged the Caribbean and Latin America into one region and combined the USA, Canada, Australia and New Zealand with northwestern Europe. Thus, the regions were: (1) Northwestern countries (Switzerland, Andorra, Austria, Belgium, Denmark, Finland, France, Germany, the UK, Iceland, Ireland, Liechtenstein, Luxemburg, Monaco, the Netherlands, Norway, Sweden, USA, Canada, Australia and New Zealand); (2) sub-Saharan Africa; (3) Southern Europe (Spain, Portugal, Italy, Greece, Malta and San Marino); (4) Latin America/Caribbean; (5) Southeastern Asia; (6) Eastern Europe/Central Asia; and (7) Northern Africa/Middle East. The SHCS collects information on ethnicity, categorized as White, Black, Asian and Hispano-American. Because there was a congruent picture between nationality and ethnicity in five out

of the seven regions described above (>96% of participants), we did not analyse the data for ethnicity separately. Demographic and clinical characteristics at inclusion were analysed for three calendar periods (1996–1999, 2000–2003 and 2004–2008) to determine trends over time. ALOX15 Cox proportional hazards models were fitted to examine the effects of region of origin, gender, age, education, IDU, clinical HIV disease stage and treatment status on the probability of ceasing to participate in the SHCS. CD4 cell count was also fitted as a time-updated covariable. Because of evidence of an interaction [likelihood ratio test (LRT) P<0.001] between region of origin and gender, we analysed the risk for LTFU separately for women and men. Data from the survey on SHCS participation were analysed using logistic regression. Because the group of former participants was very small (3.

2e). Although the above studies ascertained the formation of free radicals during PCD in Xcg, it was not clear whether these radicals are the cause or the effect of PCD. To answer this question, the effects of the ROS scavengers DMSO, glutathione

(GSH), nPG, and catalase on PCD were tested. Cell survival almost doubled in the presence of DMSO (0.25–0.5%) compared with the control at the end of a selleck chemicals llc 96-h incubation period and the increase was found to be statistically significant (P≤0.05) (Fig. 3a). However, the increase in survival was not found to be significantly affected by an increase in the DMSO concentration (P≤0.05). When GSH was added to PIM, a concentration-dependent increase in cell survival was observed when assayed at 96 h of incubation and PCD was completely inhibited with 10 mM GSH (Fig. 3b). As for GSH, PCD was also significantly abolished with 100 μM nPG (Fig. 3c) and 500 U mL−1 of catalase (Fig. 3d). No growth was observed at higher concentrations of GSH or nPG and both were found to be more effective than DMSO in inhibiting PCD. Caspase-3 biosynthesis was also found to be lower in cells grown in the presence of these ROS scavengers (Fig. 3e). In comparison with PIM-grown Xcg cells, the caspase-3 band intensity was 14%, 25%, 53%,

and 57% in cells grown in PIM in the presence of GSH (10 mM), DMSO (0.5%), nPG (100 μM), and catalase (500 U mL−1), respectively. The inhibition of caspase-3 expression Pexidartinib by DMSO (0.5%) or GSH (10 mM) was quite prominent compared with nPG or catalase.

This effect may be due to a difference in the mechanism of action of different ROS scavengers. Caspase-3 activity decreased by 15%, 10%, and 20% in Xcg cells grown in PIM 4-Aminobutyrate aminotransferase in the presence of GSH (10 mM), nPG (100 μM), and catalase (100 U mL−1), respectively, as compared with Xcg cells grown in PIM alone (Fig. 3f). Caspase-3 activity in Xcg cells grown in PIM in the presence of DMSO (0.5%) was negligible (data not shown). When a PNIM-grown Xcg cell lysate was exposed to H2O2, the level of caspase activity increased in a concentration-dependent manner, as evidenced by the observed increase in the intensity of fluorescence (Fig. 4a). Therefore, these findings indicate that H2O2 is involved in both intercellular and intracellular communication of the PCD signal in Xcg. No H2O2 could be detected by scopoletin assay in the Xcg cells grown in PIM in the presence of 500 μM 2,4-dinitrophenol (DNP) (Fig. 4b). When Xcg cells were grown in PIM with a sublethal concentration of DNP, cell survival increased by one log cycle (Fig. 4c). Figure 4d shows the effect of the addition of nalidixic acid (DNA gyrase inhibitor) to Xcg culture in PIM. The minimum inhibitory concentration of nalidixic acid for Xanthomonas sp. has been reported to be around 8–16 μg mL−1 (Pruvost et al., 1998). The results show that the addition of nalidixic acid at sublethal concentrations (0.8 and 1.

When the monolayer is not disrupted, the recovered CFU mL−1 should remain essentially constant over the same

time course. The S. Typhimurium 14028s (black diamonds) and S. Typhimurium 14028s ΔsopD2∷FRT (NT060) (white circles) showed a slight decline over the time course of the assay, suggesting that the monolayer integrity was not significantly affected by these strains (Fig. 3). In contrast, CFU mL−1 of S. Typhi STH2370 abruptly decreased until they became undetectable, strongly suggesting that gentamicin leaked due to a monolayer disruption (white squares). When S. Typhi was complemented with sopD2STM gene (in the pNT007 plasmid, see Materials and methods) and used to infect the monolayer, we observed that the corresponding ABT-263 chemical structure CFU mL−1 showed a sharp difference with the otherwise isogenic wild-type strain resembling the S. R428 supplier Typhimurium phenotype (black triangles). The CFU mL−1 numbers from infected cells with S. Typhi carrying the empty plasmid (pCC1) showed no differences with respect to the wild-type strain (data not shown). It has been reported that sopD2 contributes to the synthesis of Sifs, lipid filaments essential for S. Typhimurium intracellular

proliferation (Brumell et al., 2003; Jiang et al., 2004; Birmingham et al., 2005). When we performed a gentamicin protection assay, we observed that S. Typhi sopD2STM showed a significant decrease of CFU recovered from HEp-2-infected monolayers compared with the wild-type strain (Fig. 4). In contrast, S. Typhi sopD2STM showed similar invasion levels compared with S. Typhimurium 14028s ΔsopD2∷FRT (NT060) (P=0.13749). The results suggest that loss of SopD2 function in the serovar Typhi contributes to the bacterial intracellular proliferation in human epithelial cells. In the process of adaptation to humans, bacterial genes no longer compatible with the lifestyle of facultative

pathogens within the host are selectively inactivated. These inactivated genes are called ‘antivirulence genes’ and their loss of function results in the adaptation to a given host (Maurelli, 2007). Salmonella enterica serovar Typhi is a facultative bacterial pathogen that has accumulated a large number of pseudogenes (approximately 5% of the genome), over 75% of which have completely lost their function (McClelland et al., 2004; selleck chemical Dagan et al., 2006). Compared with free-living organism genomes, facultative pathogens harbor several pseudogenes and a gene population structure that promotes the maintenance of specific mutations. In contrast to free-living bacteria (large genomes, a great diversity of functional genes and low percentage of laterally transferred genes) and obligate parasites (extremely reduced genomes), S. Typhi represents an intermediate step exhibiting some genome erosion directed to inactivation and loss of detrimental or nonessential functions for its environment, i.e. the host (Ochman & Moran, 2001).

The assessment and subsequent recommendations are based on limited RCT data and PK interaction studies with available DAAs. ARV regimens should be selected or modified to suit the planned hepatitis C treatment. If DAAs are not being considered, standard first-line ART can be used: efavirenz, ritonavir-boosted

atazanavir, ritonavir-boosted darunavir, or raltegravir with TDF/FTC. Didanosine (increased intracellular didanosine levels and risk of toxicity with ribavirin), d4T (increase in risk of mitochondrial toxicity with ribavirin), and ZDV (overlapping toxicity with PEG-IFN and ribavirin) are contraindicated check details [64]. Some retrospective studies have shown abacavir to be associated with a decreased response to PEG-IFN/RBV therapy, possibly due to intracellular reductions in ribavirin level. However, factors including non-weight-based RBV dosing and differential baseline HCV VLs have made these data difficult to interpret. A recent study suggested no find more negative interaction when weight-based

ribavirin was utilised. Nevertheless, caution should be applied when abacavir is to be used with a ribavirin dose of ≤ 1000 mg or ≤ 13.2 mg/kg [65]. When DAAs are chosen, some restriction on first-line ARV choice exists due to drug–drug interactions. Boceprevir (BOC) and telaprevir (TPV) are currently licensed DAAs for the treatment of hepatitis C genotype 1 infection, and are substrates and inhibitors of cytochrome P (CYP) 3A4/5 and p-glycoprotein (p-gp), and therefore interact with several ARVs. Boceprevir is also metabolised by aldo-ketoreductase. Resveratrol When using TPV and BOC, only certain ARV agents are recommended for routine use due to DDI concerns (see Table 8.1). Choice of available, safe third

agents differs with use of BOC and TPV. From the limited data and drug–drug interaction studies, we recommend that if BOC is to be used, raltegravir with TDF/FTC should represent first-line ART in the presence of wild-type HIV. For TPV, we recommend that standard-dose ritonavir-boosted atazanavir or raltegravir (RAL) should be used – efavirenz can also be used but TPV dose needs to be increased to 1125 mg tds. Alternative ARVs when treating with either boceprevir or telaprevir are etravirine, rilpivirine and maraviroc, based on available pharmacokinetic (PK) data [66–68]. Multiple DAAs are currently in Phase III trials in coinfected patients.

80 with Sa113 from meat products and at minor similarity level with other two meat isolates. The remaining meat isolates grouped in different subgroups, all within group 2, which also included the remaining fish and salad isolates. In conclusion, our Ibrutinib nmr results support the idea of an early separation of L. garvieae population into two independent genomic lineages. Subsequently, the environmental stimuli of

a specific niche could have exerted a selective pressure favoring the emergence of several independent genotypes. It appears plausible that genomic flux within the dispensable genome, recombination events between genetically distinct strains during mixed colonization and/or gene (in)activation could have governed the bacterial adaptation to different habitats. Recently, we carried out the complete genome sequencing of one strain of dairy origin and one strain isolated from fish, belonging to ‘meat-group’ (Ricci et al., 2012). Whole-genome comparison between these and other L. garvieae available complete genomes, together with multilocus sequence typing (MLST) experiments are in progress in our laboratory for a deeper understanding of the

evolutionary history and the global complexity of this bacterial species. This work was supported by ‘Post genomica batterica per la qualità e la sicurezza degli alimenti’ project from the Lombardy region (Italy). We thank Dr S. Guglielmetti for a critical reading of the manuscript STK38 and for his useful Alpelisib molecular weight suggestions. “
“Interspecies bacterial communication is mediated by autoinducer-2, whose synthesis depends on luxS. Due to the apparent universality

of luxS (present in more than 40 bacterial species), it may have an ancient origin; however, no direct evidence is currently available. We amplified luxS in bacteria isolated from 25- to 40-million-year-old amber. The phylogenies and molecular clocks of luxS and the 16S rRNA gene from ancient and extant bacteria were determined as well. Luminescence assays using Vibrio harveyi BB170 aimed to determine the activity of luxS. While the phylogeny of luxS was very similar to that of extant Bacillus spp., amber isolates exhibited unique 16S rRNA gene phylogenies. This suggests that luxS may have been acquired by horizontal transfer millions of years ago. Molecular clocks of luxS suggest slow evolutionary rates, similar to those of the 16S rRNA gene and consistent with a conserved gene. Dendograms of the 16S rRNA gene and luxS show two separate clusters for the extant and ancient bacteria, confirming the uniqueness of the latter group. Interspecies bacterial communication, or quorum sensing (QS), is mediated by autoinducer-2 (AI-2), a furanosyl borate diester (Schauder et al., 2001). Synthesis of AI-2 depends on luxS, which is the product of S-ribosylhomocysteine lyase. luxS was first identified in Vibrio harveyi, Escherichia coli, and Salmonella typhimurium, and its expression has been associated with virulence in E.

All participants provided written informed consent and received a modest fee. The stimulus configuration is shown in Fig. 1. It consisted http://www.selleckchem.com/products/Everolimus(RAD001).html of two checkerboard

stimuli located 2° above and on either side of a fixation spot at horizontal eccentricities of 2.5° and 7.9°, respectively. The size of the inner checkerboards was 3.5° × 3.5°, with a spatial frequency of 0.7 cycles per degree; the size of the outer checkerboards was 4.7° × 4.7°, with a spatial frequency of 0.5 cycles per degree (Fig. 1). The larger size of the outer stimuli was chosen to adjust visual stimuli for the reduction in visual cortical area devoted to peripheral space (Adams & Horton, 2003; Frey et al., 2013). Dark checks had a luminance of 0.1 cd/m2, and white checks had a luminance of 118.2 cd/m2. The refresh rate of the monitor (model VP2655; ViewSonic, Walnut,

CA, USA) was set to 60 Hz, and on every refresh the checkerboard pattern of each stimulus either remained constant or was inverted as determined by a binary m-sequence of order 7 (e.g. (Sutter, 2000; Schmid et al., 2009). The binary m-sequence technique controls the inversion of the checkerboards displayed in each stimulus location by using GDC-0973 mouse a pseudo-random sequence, which ensures that inversions in one location are statistically independent from the inversions in all other stimulus locations. Cortical evoked responses are then obtained by cross-correlation of the continuous EEG data around stimulus reversals with the checkerboard reversal sequence. An order of 7 indicates that each sequence was 27 = 128 monitor refresh cycles (i.e. 2.1 s) long. This duration is sufficient to fit four evoked responses of duration 500 ms. In half of the trials, we used this sequence, and in the other half we usedits inverse. Each trial was 2.95 s in length; however, the m-sequence used for estimating the evoked cortical response was only 2.1 s in length. In order to minimise stimulus onset

artefacts, L-gulonolactone oxidase we used another random sequence for the first 850 ms of each trial, and this time-frame was excluded from further analysis. For the experimental task, we overlaid each checkerboard with a central red ‘X’ (task stimulus). At the beginning of each block of 20 trials, participants were instructed to simultaneously attend to two of the checkerboards, and count how many times their task stimuli disappeared at the same time. This ensured that participants did not have to switch attention on each trial. Before each experimental trial, the two attended checkerboards were cued again, and, after a random interstimulus interval of 800–1200 ms, the experimental trial was started. Participants were instructed to ignore the uncued checkerboards, as task stimuli could also disappear in the uncued locations.

Simulated patients (SPs) were used

to evaluate pharmacy staff performance. Ten SPs were recruited and trained. Eight were selected to participate in the study and each was allocated one scenario to perform. The SPs made covert visits to each participating pharmacy over a four-week period. Each visit was audio-taped and the SP completed a data collection form, which included their overall satisfaction with the consultation and staff members, in terms of professionalism. This was completed immediately after leaving each pharmacy. Audio-taped consultations were scored by three members of the research team and a consultation score was derived from components which Selumetinib mw included information gathering and advice provision using criteria established by the MCP and modelled on an adapted form of the Calgary Cambridge communications skills model2. Both sets of data were then entered into SPSS and a 10% accuracy check performed. Descriptive see more statistics were generated. Ethical approval was received from the North of Scotland Research Ethics Committee. In total, 72 SP visits were made to the 18 pharmacies. Each pharmacy received four visits, one for each scenario. Recordings were available for 68 consultations. Only one of the SP visits was detected

by pharmacy staff. SP visits for Sirolimus back pain achieved the highest consultation scores with higher scores indicating greater compliance with MCP recommendations (Table 1). The management of sore throat achieved the lowest levels of compliance with the MCP recommendations. Most SP visits achieved high scores for the professionalism with which the consultation had been managed

and around a third of SP visits were scored as being of an exceptional interaction in terms of their overall management. Table 1: Simulated Patients’ Consultation scores and ratings of professionalism and overall satisfaction with minor ailment consultations Scenario Consultation score Average (range 0 to1) General professionalism (completely satisfied/satisfied) n (%) Overall satisfaction (exceptional interaction) n (%) Back pain 0.69 (0.2 to1) 18 (100) 6 (36.8) Eye discomfort 0.51 (0 to 1) 16 (89.5) 6 (36.8) Gastro-intestinal upset 0.53 (0.2 to 0.9) 18 (100) 6 (36.8) Sore throat 0.45 (0 to 1) 17 (93.3) 5 (26.7) The consultation score reflected pharmacy staff members’ communication performance during these consultations. The results suggest that there is scope for improvement with regard to communication behaviour during consultations for the management of minor ailments. Sub-optimal communication may be due to lack of training, knowledge, or may reflect pharmacy staff attitudes towards information elicitation from consumers.