The organic

layers were combined and dried using an evaporator at 55 °C. The n-butanol extract was suspended in distilled water, and applied to an MCI GEL CHP20P (75–150 μm) column equilibrated with distilled water. The column was washed extensively with distilled water and then eluted with stepwise gradients of aqueous methanol (30, 50, 70, 90 and 100%, v/v). Each fraction was collected and the antibacterial activity was evaluated using the agar diffusion method (Al-Bayati, 2009). Bacillus subtilis CGMCC 1.1470 was used as the indicator strain. The active fractions eluted with 50% and 70% methanol in water were combined and concentrated. This material was then purified using a preparative HPLC system (Dalian Elite, Dalian, China) equipped with a YMC-pack DOS-A C18 (5 μm, 250 × 20 mm) column. The mobile phase consisted of Milli-Q water containing 0.02% trifluoroacetic selleck inhibitor acid and acetonitrile. A triphasic linear gradient of 28–28% acetonitrile (15 min), 28–35% acetonitrile Epacadostat mw (5 min) and 35–45% acetonitrile (40 min) was used for elution at a flow rate of 8 mL min−1. The elution was detected at 210 nm. All isolatable peaks were collected and assessed for antimicrobial activity. The fractions with antimicrobial

activity were vacuum evaporated to dryness. The stability of CFS against heat, pH variation and enzyme treatments was investigated. All experiments were conducted in triplicate. For the heat treatment, CFS was incubated at 40, 60, 80 and 100 °C for 2 h. For pH stability,

CFS was adjusted to pH 1.0–12.0 with HCl or NaOH and held overnight at 4 °C. The treated CFS was neutralized to pH 7.0 before performing an antimicrobial activity assay. To determine its stability against degradative enzymes, CFS was treated with several enzymes at a final concentration of 1 mg mL−1 (Lee et al., 2007). Before the enzymes were added, CFS was adjusted to pH 2.0 for pepsin and pH 7.5 for proteinase K, trypsin and lipase. The reaction mixtures were incubated at 37 °C Cediranib (AZD2171) for 2 h. After the different treatments, the remaining antimicrobial activities of the CFS samples were assessed using the agar diffusion method (Al-Bayati, 2009). Bacillus subtilis CGMCC 1.1470 was used as the indicator strain. The amino acid analyses were carried out using the advanced Marfey’s method with LC/MS. The FDLA derivatives of the purified antibiotics were prepared as described by Fujii et al. (1999). The separation of the l- and d-FDLA derivatives was performed on a ZORBAX SB-C18 (3.5 μm, 150 × 2.1 mm) column with the mobile phase: 20 mM NH4Ac water solution and acetonitrile. A triphasic linear gradient of 10–20% acetonitrile (5 min), 20–50% acetonitrile (35 min) and 50–90% acetonitrile (5 min) was applied at a flow rate of 0.2 mL min−1. The elution pattern was monitored at 340 nm.

Alternatively, TraB might recruit other chromosomally

encoded proteins for the transfer process. 1. How to cross the PG barrier? A TraB–eGFP fusion was localized at the hyphal tip, suggesting that the selleck chemical tips of the mycelium are involved in conjugation (Reuther et al., 2006a). Also, TraB was shown to bind isolated PG (Vogelmann et al., 2011a). Because TraB itself does not have a PG-lysing activity (Finger and Muth, unpublished), it is possible that TraB interacts with chromosomally encoded PG hydrolases at the tip to direct fusion of the PG layers of donor and recipient. 2. How to cross membranes of donor and recipient? In contrast to FtsK that is found in both compartments during cell division, TraB is present only in the donor mycelium. Therefore, the TraB pore has to traverse two membranes (one from the donor, one from the recipient) or the two membranes have to fuse. For SpoIIIE that mediates translocation of the chromosome into the forespore during Bacillus sporulation, a membrane fusing activity has been reported (Sharp & Pogliano, 2003). Therefore, it is tempting to speculate that also TraB might have a membrane

fusing activity allowing formation of a pore structure to the recipient. 3. How to translocate a circular covalently closed plasmid molecule? During cell division or sporulation, Selleck Stem Cell Compound Library the septum closes, while chromosomal DNA is already present, allowing FtsK to assemble at both chromosomal arms to translocate the DNA. DNA translocation causes topological stress to the DNA, which has to be relieved by topoisomerases. The interaction of E. coli FtsK with topoisomerase

L-gulonolactone oxidase IV has been reported (Espeli et al., 2003). However, it is still unclear, how the remaining end of the circular chromosome becomes translocated through the membrane and fusion of the two FtsK hexamer structures has been postulated (Burton et al., 2007). During Streptomyces conjugation, the situation is even more complex. The translocase TraB is definitely present only on the donor site of the mating hyphae, and a mechanism translocating a circular double-stranded DNA molecule is not very plausible. Because the plasmid DNA is not processed during TraB binding at clt, one has to propose involvement of an additional enzymatic activity, for example, a topoisomerase, which might produce a linear molecule that can be transported through the TraB pore. 4. How to pass the septal cross-walls in the recipient mycelium? Crossing the septal cross-walls during intramycelial plasmid spreading seems to be an even more challenging task compared to the primary DNA transfer at the hyphal tip. It involves, in addition to TraB, several Spd proteins. The structure of the Streptomyces septal cross-walls has not been elucidated, and it is not clear whether preexisting channel structures in the cross-walls connect the compartments of the substrate mycelium (Jakimowicz & van Wezel, 2012).

Alternatively, TraB might recruit other chromosomally

encoded proteins for the transfer process. 1. How to cross the PG barrier? A TraB–eGFP fusion was localized at the hyphal tip, suggesting that the Trichostatin A tips of the mycelium are involved in conjugation (Reuther et al., 2006a). Also, TraB was shown to bind isolated PG (Vogelmann et al., 2011a). Because TraB itself does not have a PG-lysing activity (Finger and Muth, unpublished), it is possible that TraB interacts with chromosomally encoded PG hydrolases at the tip to direct fusion of the PG layers of donor and recipient. 2. How to cross membranes of donor and recipient? In contrast to FtsK that is found in both compartments during cell division, TraB is present only in the donor mycelium. Therefore, the TraB pore has to traverse two membranes (one from the donor, one from the recipient) or the two membranes have to fuse. For SpoIIIE that mediates translocation of the chromosome into the forespore during Bacillus sporulation, a membrane fusing activity has been reported (Sharp & Pogliano, 2003). Therefore, it is tempting to speculate that also TraB might have a membrane

fusing activity allowing formation of a pore structure to the recipient. 3. How to translocate a circular covalently closed plasmid molecule? During cell division or sporulation, BMS-354825 concentration the septum closes, while chromosomal DNA is already present, allowing FtsK to assemble at both chromosomal arms to translocate the DNA. DNA translocation causes topological stress to the DNA, which has to be relieved by topoisomerases. The interaction of E. coli FtsK with topoisomerase

CYTH4 IV has been reported (Espeli et al., 2003). However, it is still unclear, how the remaining end of the circular chromosome becomes translocated through the membrane and fusion of the two FtsK hexamer structures has been postulated (Burton et al., 2007). During Streptomyces conjugation, the situation is even more complex. The translocase TraB is definitely present only on the donor site of the mating hyphae, and a mechanism translocating a circular double-stranded DNA molecule is not very plausible. Because the plasmid DNA is not processed during TraB binding at clt, one has to propose involvement of an additional enzymatic activity, for example, a topoisomerase, which might produce a linear molecule that can be transported through the TraB pore. 4. How to pass the septal cross-walls in the recipient mycelium? Crossing the septal cross-walls during intramycelial plasmid spreading seems to be an even more challenging task compared to the primary DNA transfer at the hyphal tip. It involves, in addition to TraB, several Spd proteins. The structure of the Streptomyces septal cross-walls has not been elucidated, and it is not clear whether preexisting channel structures in the cross-walls connect the compartments of the substrate mycelium (Jakimowicz & van Wezel, 2012).

Proteins were separated on 15% SDS-PAGE. The fluorescent coumarin-labeled proteins were monitored under UV light. The effect of various heavy metals on the expression of the ctsR operon was profiled by qRT-PCR analysis. Defined media (Townsend & Wilkinson, 1992) were used to simulate a heavy-metal-deficient environment. Staphylococcus aureus strain SH1000 was grown overnight at 37 °C in TSB. The following day, the cells were collected and suspended in defined media. The culture was then grown overnight. The culture was diluted 1:100 in fresh defined media and grown to OD600 nm

at 0.4. CuSO4 (200 μM), ZnSO4 (100 μM), CoCl2 (200 μM) or CdCl2 (100 μM) was added and the cells were grown for an additional 1 h. One Selleck Navitoclax culture was grown without excess metal ions. After 1 h, the cells were collected and total RNA was extracted using the RNeasy Protect Bacteria Mini Kit (Qiagen) and quantified by 260 nm spectrophotometry

(Nanodrop, Thermo Scientific). One microgram of total RNA was converted to cDNA using AG-014699 ic50 the High Capacity RNA-to-cDNA Kit (Applied Biosystems) following manufacturer’s protocols. qRT-PCR was then performed using the DyNAmo Flash SYBR Green qPCR Kit (Thermo Scientific) using gene-specific primers (Table 2), and data were collected using the ABI 7300 Real-Time PCR System (Applied Biosystems). Up- or down-regulation was normalized

against the 16S rRNA gene and then against the uninduced cultures. All reactions were run in triplicate on the same cultures. A bacterial two-hybrid system was constructed using the pB2H∆α and pB2∆ω vectors as described by Borloo et al. (2007). DNA fragments Oxalosuccinic acid of the upstream and downstream regions of ctsR or mcsB were amplified from genomic DNA using primer pairs as shown on Table S1. The PCR product was first cloned in frame into the PCR2.1 vector (Invitrogen) and subsequently into the SphI and BamHI sites of pB2H∆α. The mcsA gene was amplified from genomic DNA using primers mcsA-F and mcsA-B (Table S1). The PCR product was cloned in frame into vector PCR2.1 and subsequently into the BamHI and SphI sites of pB2H∆ω. To test the function of the CXXC cysteines from McsA, site-directed mutagenesis was performed to replace Cys residues to Ala at the N-terminus of McsA as described above. The PCR product was first cloned in frame into the PCR2.1 vector, and mutation was confirmed by DNA sequencing. The fragment corresponding to mutated protein was gel-purified and subcloned into the SphI and BamHI site of pB2H∆ω. To co-express the fusion proteins, an E. coli MC1061 containing pB2H∆ω-mcsA or pB2H ∆ω-∆mcsA was transformed with pB2H∆α-ctsR or pB2H∆α-mcsB.

Generally the number of HIF-1α-positive cells is strongly correlated with the number of blood vessels in RA synovial tissue and with inflammatory EC infiltration.[44, 45] Some data demonstrate that HIF-1α causes a noticeable reduction in the ability of smooth muscle cells to migrate and adhere to extracellular matrices. Moreover, findings by Kennedy et al. in 2010 indicate the presence

of unstable vessels in inflamed joints is correlated with hypoxia, insufficient ECs/pericyte interactions, and increased DNA damage. These changes may contribute to persistent hypoxia in the inflamed joint to further manage this unstable microenvironment.[10] In fibroblast-like synoviocytes (FLS), hypoxia-induced MMP-3 expression is exclusively regulated by HIF-1α, while hypoxia-induced MMP-1 or IL-8 Cell Cycle inhibitor expression appears to have salvage pathways other than the HIF-1α pathway.[46] This demonstrated that migration and invasion of FLSs are critical in the pathogenesis of RA. Li and colleagues in their Apitolisib current study observed that RA-FLSs exposed to hypoxic conditions experienced epithelial-mesenchymal transition (EMT), with increased cell migration and invasion. In this study hypoxia-induced EMT was accompanied by increased HIF-1α expression and activation of Akt. Therefore activation of the PI3K/Akt/HIF-1α pathway plays a pivotal role in mediating

hypoxia-induced EMT transformation and invasion of RA-FLSs under hypoxia status.[47] As we know, the combination of hypoxia and IL-17A factor promote the migration and invasion of FLSs, which are critical for the pathogenesis of RA. However, the biochemical pathways regulating IL-17A combined with hypoxia are not well isothipendyl defined, but recent observations suggest a synergetic effect of IL-17A and hypoxia that might contribute to the migration and invasion of RA-FLSs by up-regulating the expression of MMP-2 and MMP-9 by activation of the NF-κB/HIF-1α pathway.[48] Alternatively, hypoxia is thought to drive an increase in the synovial angiogenesis process that occurs in RA, through expression

of a number of angiogenic factors, including VEGF, Ang, HGF and FGF-2. Here, HIF-1α and HIF-2α are also essential in regulating transcription of the VEGF gene and finally increased vascularity in the inflammation region. This process promotes further infiltration of inflammatory cells and production of inflammatory mediators, perpetuating synovitis.[36, 44, 49] Notch signaling pathways are crucial for angiogenesis and EC fate. In a recent study, the effect of hypoxia on Notch-1 signaling pathway components and angiogenesis in inflammatory arthritis synovial tissue was examined. The results indicate that Notch-1 is expressed in synovial tissue and that increased Notch-1 intracellular domain (NICD) expression is associated with low in vivo tissue oxygen levels. Furthermore, Notch-1/HIF-1α interactions via VEGF/Ang-2, mediate hypoxia-induced angiogenesis and invasion in inflammatory arthritis.

Generally the number of HIF-1α-positive cells is strongly correlated with the number of blood vessels in RA synovial tissue and with inflammatory EC infiltration.[44, 45] Some data demonstrate that HIF-1α causes a noticeable reduction in the ability of smooth muscle cells to migrate and adhere to extracellular matrices. Moreover, findings by Kennedy et al. in 2010 indicate the presence

of unstable vessels in inflamed joints is correlated with hypoxia, insufficient ECs/pericyte interactions, and increased DNA damage. These changes may contribute to persistent hypoxia in the inflamed joint to further manage this unstable microenvironment.[10] In fibroblast-like synoviocytes (FLS), hypoxia-induced MMP-3 expression is exclusively regulated by HIF-1α, while hypoxia-induced MMP-1 or IL-8 selleckchem expression appears to have salvage pathways other than the HIF-1α pathway.[46] This demonstrated that migration and invasion of FLSs are critical in the pathogenesis of RA. Li and colleagues in their Pirfenidone solubility dmso current study observed that RA-FLSs exposed to hypoxic conditions experienced epithelial-mesenchymal transition (EMT), with increased cell migration and invasion. In this study hypoxia-induced EMT was accompanied by increased HIF-1α expression and activation of Akt. Therefore activation of the PI3K/Akt/HIF-1α pathway plays a pivotal role in mediating

hypoxia-induced EMT transformation and invasion of RA-FLSs under hypoxia status.[47] As we know, the combination of hypoxia and IL-17A factor promote the migration and invasion of FLSs, which are critical for the pathogenesis of RA. However, the biochemical pathways regulating IL-17A combined with hypoxia are not well Silibinin defined, but recent observations suggest a synergetic effect of IL-17A and hypoxia that might contribute to the migration and invasion of RA-FLSs by up-regulating the expression of MMP-2 and MMP-9 by activation of the NF-κB/HIF-1α pathway.[48] Alternatively, hypoxia is thought to drive an increase in the synovial angiogenesis process that occurs in RA, through expression

of a number of angiogenic factors, including VEGF, Ang, HGF and FGF-2. Here, HIF-1α and HIF-2α are also essential in regulating transcription of the VEGF gene and finally increased vascularity in the inflammation region. This process promotes further infiltration of inflammatory cells and production of inflammatory mediators, perpetuating synovitis.[36, 44, 49] Notch signaling pathways are crucial for angiogenesis and EC fate. In a recent study, the effect of hypoxia on Notch-1 signaling pathway components and angiogenesis in inflammatory arthritis synovial tissue was examined. The results indicate that Notch-1 is expressed in synovial tissue and that increased Notch-1 intracellular domain (NICD) expression is associated with low in vivo tissue oxygen levels. Furthermore, Notch-1/HIF-1α interactions via VEGF/Ang-2, mediate hypoxia-induced angiogenesis and invasion in inflammatory arthritis.

In this study, a soil-borne, glyphosate-resistant bacterium was selected and identified as Enterobacter. The EPSPS in this strain was found to have p38 protein kinase been altered to a resistant one. A total of 42 differentially expressed genes (DEGs) in the glyphosate were screened using microarray techniques. Under treatment, argF, sdhA, ivbL, rrfA-H were downregulated, whereas the transcripts of speA, osmY, pflB, ahpC, fusA, deoA, uxaC, rpoD and a few ribosomal protein genes were upregulated. Data were verified by quantitative real-time PCR on selected

genes. All transcriptional changes appeared to protect the bacteria from glyphosate and associated osmotic, acidic and oxidative stresses. Many DEGs may have the potential to confer resistance to glyphosate alone, and some may be closely related to the shikimate pathway, reflecting the complex gene interaction network for glyphosate resistance. “
“Yersinia polynucleotide phosphorylase (PNPase), a 3’-5’ exoribonuclease, has been shown to affect growth during several stress responses. In E. coli, PNPase is one of the subunits of a multi-protein complex known as the degradosome, but also has degradosome-independent

functions. The carboxy–terminus of E. coli ribonuclease E (RNase E) serves as the scaffold upon which PNPase, enolase (a glycolytic enzyme), and RhlB helicase all have been shown to bind. In the yersiniae, only PNPase has thus far been shown to physically interact with RNase BEZ235 E. We show by bacterial two-hybrid and co-immunoprecipitation assays that RhlB and enolase also interact with RNase E. Interestingly, although PNPase is required for normal growth at cold temperatures, assembly of the yersiniae degradosome was not required. However, degradosome assembly was required for growth in the presence of reactive oxygen species. These data suggest that while the Y. pseudotuberculosis PNPase plays a role in the oxidative stress response through a degradosome-dependent mechanism, PNPase’s this website role during cold stress is degradosome-independent.

2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved “
“To examine why we failed in direct sequencing of rRNA gene internal transcribed spacer (ITS) in Pleurotus nebrodensis, we obtained monokaryons of P. nebrodensis (00489 and 00491) using a protoplast monokaryonization technique. PCR products of ITS amplifications were sequenced. There was a base pair insertion/deletion difference between the two nuclei of P. nebrodensis that led to failure in direct sequencing. Internal transcribed spacer regions (ITS1, ITS2, and 5.8S rRNA gene) of the nuclear ribosomal repeat are widely used in fungal systematics and phylogeny (Gardes & Bruns, 1993; Kårén et al., 1997; Cooke et al., 2000; Manter & Vivanco, 2007; Nilsson et al., 2009).

To identify potential spa type changes, the spa types from the 319 patients with repeated

MRSA occurrence were analyzed (Fig. 1). Ninety-four percent of the patients had MRSA of only one spa type while 20 patients had two spa types. No one had more than two types. Of these 20 patients, seven had spa types so different from each other that they were considered to be two independent MRSA acquisitions. The remaining 13 patients had two spa types that were closely related. The gender and age, spa type and body site of MRSA isolate, and time between sampling for these isolates is shown in Table 1. The age of the patients varied from 19 to 90. Between two and 17 MRSA isolates were recovered from each patient. The time from the first to the last recording of MRSA was Navitoclax solubility dmso between 0 (simultaneously) and 22 months. Out of 88 isolates, the largest group (40) was from skin and soft tissue infection, 23 samples from the nose and 13 from the throat. To further evaluate whether the paired isolates from the same patient were clonally related, a multiple-locus variable number tandem repeat analysis (MLVA) was performed,

based on the method described by Schouls et al. (2009). The discriminatory power of this MLVA is the same as pulsed-field gel electrophoresis and MLVA types can be clustered into MLVA complexes, which coincide with MLST clonal DNA Damage inhibitor complexes (Schouls et al., 2009). In the present study, the MLVA was performed with seven primer pairs (spa-primers were excluded). For PCR, fluorescent dyes were omitted. PCR bands were fragmented on a Qiaxcel

System (Qiagen, Hilden, Germany) and band sizes were determined using the qiaxcel biocalculator software. MLVA analysis was performed on the first isolate of Ancestor and Variant spa types. Thirteen of 319 patients Adenosine triphosphate (4%) with a total of 30 MRSA isolates had two spa types with changes that could be assigned to mutational events, suggesting clonal microevolution in the spa repeat region. Twelve different events of spa type change were found; one change was found in two individuals (Table 2). MLVA analysis confirmed that all pairs of isolates were clonally related. All pairs of Ancestor and Variant exhibited identical band patterns except one case with a single band difference (data not shown). The most common mutational event detected was a deletion of spa repeats (10 events), followed by repeat duplication (three events) and point mutation (one event) (Table 2). This was in agreement with the changes found by Kahl et al. (2005) and Sakwinska et al. (2010). Between one and nine repeats were deleted. In two cases (patients 7 and 9), the deletion seemed to be not of a repeat but of 24 bps spanning two repeats, thereby creating a new repeat from parts of the two original repeats. Three changes probably involved repeat duplication. In the first case, spa types t005 and t1276 were involved (patient 3; Table 2). Because t005 is often found in Copenhagen, we consider it to be the Ancestor.

Researchers suppose that the first choice for treating both neutropenia and arthritis is methotrexate, which is safe, effective and well tolerated in these patients.[2] Many studies suggest that application of rituximab is useful KU-60019 in the treatment of FS, while other researchers have found a different result.[3] Controlled trials of different

treatment modalities are not available because of the rarity of this syndrome. Splenectomy has produced a long-term hematologic response in 80% of patients but is usually reserved at the end of the treatment algorithm for treatment-resistant cases.[4] In some patients with FS, the presence of antibodies against neutrophils has Palbociclib been described, which might be associated with increased neutrophil destruction. The underlying mechanism of developing neutropenia in FS is similar to that in other forms of immune-mediated neutropenia.[5] However, there are no reports of the prevalence of association between hyperthyroidism and FS. Thus, autoimmune or immunologic processes were assumed in the pathogenesis of both autoimmune thyroid diseases (Graves’ disease) and FS. It had become recognized that Th1/Th2 balance controls the immune system. From the viewpoint of imbalance,

autoimmune Graves’ disease was considered to be a Th2-type disease.[6] Systemic involvement in RA is characterized by B cell overactivity, immune complex formation and complement consumption, suggesting that Th2 cells are involved in the pathogenesis of extra-articular manifestation of FS. Therefore, regarding Th1/Th2 imbalance, it is not surprising that there is a prevalence of Graves’ disease

in FS patients. Leflunomide may exert its effects by inhibiting the mitochondrial enzyme dihydroorotate dehydrogenase, which plays a pivotal role in the synthesis of the pyrimidine ribonucleotide uridine monophosphate (rUMP). Therefore, we propose that leflunomide prevents the expansion of activated and autoimmune lymphocytes by interfering with the cell cycle progression caused by inadequate production of rUMP.[7] This study was supported by ‘The Reverse transcriptase Incubative Program for Youth Scientists of Jiangxi Province, China’ no. 20112BCB23029. There is no potential conflict of interest in this paper. “
“Aim:  To determine the prevalence, correlates and impact of shoulder pain in a population-based sample. Methods:  The North West Adelaide Health Study is a representative longitudinal cohort study of people aged 18 years and over. The original sample was randomly selected and recruited by telephone interview. Overall, 3206 participants returned to the clinic during the second stage (2004–2006) and were asked to report whether they had pain, aching or stiffness on most days in either of their shoulders.

8%; only 4% experienced bothersome side effects. Satisfaction with the pharmacist and service was strong; only 5.6% felt a physician would have been more thorough. Participants were very satisfied with their symptomatic improvement and with the service in general, albeit for a small number of conditions. Participants reported getting VE-821 cost better, and side effects were not a concern. These results are encouraging for pharmacists; however, a comparison of physician care with pharmacist care and unsupported self-care is

required to truly know the benefit of pharmacist prescribing. “
“Objectives  The objective of this study was to examine the interaction between job demands of pharmacists and resources in the form of interpersonal interactions and its association with work-related outcomes such as organizational and professional commitment, job burnout, professional identity and job satisfaction. The job demands-resources (JD-R) model served as the theoretical framework. Methods  Subjects

for the study were drawn from the Pharmacy Manpower TSA HDAC cell line Project Database (n = 1874). A 14-page mail-in survey measured hospital pharmacists’ responses on the frequency of occurrence of various job-related scenarios as well as work-related outcomes. The study design was a 2 × 2 factorial design. Responses were collected on a Likert scale. Descriptive statistics, reliability analyses and correlational and multiple regression analyses were conducted using SPSS version 17 (SPSS, Chicago, IL, USA). Key findings  The 566 pharmacists (30% response rate) who responded to the survey indicated that high-demand/pleasant encounters and low-demand/pleasant encounters occurred more frequently in the workplace. The strongest correlations

were found between high-demand/unpleasant encounters and frequency and intensity of emotional exhaustion. Multiple regression analyses indicated PD184352 (CI-1040) that when controlling for demographic factors high-demand/unpleasant encounters were negatively related to affective organizational commitment and positively related to frequency and intensity of emotional exhaustion. Low-demand/pleasant encounters were positively related to frequency and intensity of personal accomplishment. Low-demand/unpleasant encounters were significantly and negatively related to professional commitment, job satisfaction and frequency and intensity of emotional exhaustion, while high-demand/pleasant encounters were also related to frequency and intensity of emotional exhaustion Conclusion  Support was found for the JD-R model and the proposed interaction effects. Study results suggest that adequate attention must be paid to the interplay between demands on the job and interactions with healthcare professionals to improve the quality of the pharmacist’s work life. Future research should examine other types of job demands and resources.