RELATIONSHIP BETWEEN WORKPLACE HARASSMENT AND PSYCHOLOGICAL DISTRESS AMONG SALES GIRLS IN PAKISTAN: MODERATING ROLE OF COPING

The purpose of this study is to find out the relationship between Workplace Harassment, Coping Strategies and Psychological Distress among Sales Girls in Pakistan. Data of 130 participants was collected from different shopping malls of Lahore city. The age range of the participants was 18 to 35 years (M= 22.88, SD=3.42). NAQ-R, Brief COPE Inventory and Kessler Psychological Distress Scale were used to collect data from participants. The findings indicated significant positive relationship between workplace harassment and psychological distress. Workplace harassment has significant positive relationship with problem focused coping. Avoidant emotional coping is also significantly and positively correlated with PD. Coping strategies moderate between the workplace harassment and psychological distress among sales girls. The issue of workplace harassment has broad consequences. At any organizational level workplace harassment creates unpleasant working environment and also effects on their psychological health. To manage this problem of harassment, women take support of different coping strategies. The positive relationship of avoidant coping strategy with psychological distress explains that women feel hesitate to report so they try to avoid the situation that could affect their psychological health

Probing the Putative Active Site of YjdL: An Unusual Proton-Coupled Oligopeptide Transporter from <em>E. coli</em>

<div><p>YjdL from <em>E. coli</em> is an unusual proton-coupled oligopeptide transporter (POT). Unlike prototypical POTs, dipeptides are preferred over tripeptides, in particular dipeptides with a positively charged C-terminal residue. To further understand this difference in peptide specificity, the sequences of YjdL and YdgR, a prototypical <em>E. coli</em> POT, were compared in light of the crystal structure of a POT from <em>Shewanella oneidensis</em>. Several residues found in the putative active site were mutated and the activities of the mutated variants were assessed in terms of substrate uptake assays, and changes in specificity in terms of uptake inhibition. Most strikingly, changing the YjdL specific Asp392 to the conserved Ser in YjdL obliterated the preference for a positively charged C-terminal residue. Based on this unique finding and previously published results indicating that the dipeptide N-terminus may interact with Glu388, a preliminary orientation model of a dipeptide in the YjdL cavity is presented. Single site mutations of particularly Ala281 and Trp278 support the presented orientation. A dipeptide bound in the cavity of YjdL appears to be oriented such that the N-terminal side chain protrudes into a sub pocket that opens towards the extracellular space. The C-terminal side chain faces in the opposite direction into a sub pocket that faces the cytoplasm. These data indicated a stabilizing effect on a bulky N-terminal residue by an Ala281Phe variant and on the dipeptide backbone by Trp278. In the presented orientation model, Tyr25 and Tyr58 both appear to be in proximity of the dipeptide backbone while Lys117 appears to be in proximity of the peptide C-terminus. Mutational studies of these conserved residues highlight their functional importance.</p> </div

Primers used for site-directed mutagenesis.

<p>IC₅₀ values (mM) with SEM (n ≥3) of selected dipeptides tested on YjdL and YdgR variants.</p>a<p>IC₅₀ values determined at 0.2 mM β-Ala-Lys(AMCA) and 5 min incubation.</p>b<p>IC₅₀ values determined at 0.5 mM β-Ala-Lys(AMCA) and 15 min incubation.</p

Correction: Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production.

[This corrects the article DOI: 10.1371/journal.pone.0153436.]

Activity of the Human Rhinovirus 3C Protease Studied in Various Buffers, Additives and Detergents Solutions for Recombinant Protein Production.

Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications

Schematic dipeptide orientation model for YjdL exemplified by Tyr-Lys.

<p>The cavity shape has been adapted from PepT_So (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047780#pone-0047780-g001" target="_blank">Figure 1B</a>). The labeled spheres indicate the position of the C-alpha of the corresponding residue in PepT_So. Residues behind the plane of the peptide are colored brown.</p

IC₅₀ values of C-terminal lysyl dipeptides.

<p>Forward primer sequences used to generate mutations in <i>yjdL</i> by site-directed mutagenesis. The reverse complement of the forward primers was used as the reverse primers.</p

Inhibition profiles for WT-YjdL (white), Asp392Glu (grey), and Asp392Ser (black).

<p>The cells were incubated in assay buffer pH 6.5 containing 0.5 mM β-Ala-Lys(AMCA) and 50 mM ligand. Error bars indicate SEM (n ≥3).</p

functional analyses of YdgR variants.

<p>(A) Representative western blots of YdgR mutants. (B) β-Ala-Lys(AMCA) uptake (0.2 mM, 5 min) by YdgR mutants in uptake buffer, pH 6.5. Error bars indicate SEM (n ≥3). Lys133Arg and Lys133Gln are not significantly different, P>0.05, from background levels (pTTQ18).</p