Diversity and Inclusion in the Workplace: Employee Perception and Implications for Internal Brand Management

The study explores the issues connected with diversity and inclusion in the workplace and their application in internal branding. The authors draw attention to the depth of the problematics and identify the gaps in research results concerning the meaning of diversity management as a method of creating corporate image of the organization, both externally and internally. The purpose of the research is to investigate employees’ perceptions on diversity in the workplace. It will develop insight into and knowledge of the current state of diversity in the workplace in Poland. Secondly, the results of the research are meant to help to accelerate the process of adapting and changing current management practices to meet the demands of a diversifying workforce. Besides, the authors investigate how diversity management can become an effective tool for internal branding as a crucial success factor. In the statistical study, the Mann–Whitney U test was used to assess differences between women and men. In the case of an independent variable related to the position held in the organization, the Kruskal-Wallis ANOVA analysis was used for comparison. In order to assess the relationship between nominal features, the Fisher’s exact test – the chi-squared test was applied with the Yates’s continuity correction for a 2x2 contingency table. For these tables, the contingency coefficient C of correlation was calculated. In the case of the scaled variables, to evaluate the relationship between them and the answers to the questions asked, Spearman’s rank correlation coefficients were calculated. Keywords: diversity and inclusion, diversity management, internal brand management, employer branding

Human Cysteine Cathepsins Are Not Reliable Markers of Infection by Pseudomonas aeruginosa in Cystic Fibrosis

Cysteine cathepsins have emerged as new players in inflammatory lung disorders. Their activities are dramatically increased in the sputum of cystic fibrosis (CF) patients, suggesting that they are involved in the pathophysiology of CF. We have characterized the cathepsins in CF expectorations and evaluated their use as markers of colonization by Pseudomonas aeruginosa. The concentrations of active cathepsins B, H, K, L and S were the same in P. aeruginosa-positive (19 Ps+) and P. aeruginosa-negative (6 Ps−) samples, unlike those of human neutrophil elastase. Also the cathepsin inhibitory potential and the cathepsins/cathepsin inhibitors imbalance remained unchanged and similar (∼2-fold) in the Ps+ and Ps− groups (p<0.001), which correlated with the breakdown of their circulating cystatin-like inhibitors (kininogens). Procathepsins, which may be activated autocatalytically, are a potential proteolytic reservoir. Immunoblotting and active-site labeling identified the double-chain cathepsin B, the major cathepsin in CF sputum, as the main molecular form in both Ps+ and Ps− samples, despite the possible release of the ∼31 kDa single-chain form from procathepsin B by sputum elastase. Thus, the hydrolytic activity of cysteine cathepsins was not correlated with bacterial colonization, indicating that cathepsins, unlike human neutrophil elastase, are not suitable markers of P. aeruginosa infection

Production of Cystatin C Wild Type and Stabilized Mutants

Cystatin C is produced in all nucleated cells. It has various functions and biological activities. Researchers are focused on its role in kidney diseases as a marker of glomerular filtration but also as a very important link in development of amyloid diseases. This work describes expression and purification of both wild type (wt) and stabilized form (stab 1 and 2) of wt cystatin C and amyloid-forming L68Q mutant of cystatin C. The recombinant cystatin C can be used in projects requiring pure cystatin C to examine models of dimerization and fibrils formation as well as a standard in clinical tests

DMPC Phospholipid Bilayer as a Potential Interface for Human Cystatin C Oligomerization: Analysis of Protein-Liposome Interactions Using NMR Spectroscopy

Studies revolving around mechanisms responsible for the development of amyloid-based diseases lay the foundations for the recognition of molecular targets of future to-be-developed treatments. However, the vast number of peptides and proteins known to be responsible for fibril formation, combined with their complexity and complexity of their interactions with various cellular components, renders this task extremely difficult and time-consuming. One of these proteins, human cystatin C (hCC), is a well-known and studied cysteine-protease inhibitor. While being a monomer in physiological conditions, under the necessary stimulus&mdash;usually a mutation, it tends to form fibrils, which later participate in the disease development. This process can potentially be regulated (in several ways) by many cellular components and it is being hypothesized that the cell membrane might play a key role in the oligomerization pathway. Studies involving cell membranes pose several difficulties; therefore, an alternative in the form of membrane mimetics is a very attractive solution. Here, we would like to present the first study on hCC oligomerization under the influence of phospholipid liposomes, acting as a membrane mimetic. The protein&ndash;mimetic interactions are studied utilizing circular dichroism, nuclear magnetic resonance, and size exclusion chromatography

Identification of the epitope for anti-cystatin C antibodies

Human cystatin C (hCC), like many other amyloidogenic proteins, has been shown to form dimers by exchange of subdomains of the monomeric protein. Considering the model of hCC fibrillogenesis by propagated domain swapping, it seems possible that inhibition of this process should also suppress the entire process of dimerization and fibrillogenesis which leads to specific amyloidosis (hereditary cystatin C amyloid angiopathy (HCCAA)). It was reported that exogenous agents like monoclonal antibody against cystatin C are able to suppress formation of cystatin C dimers. In the effort to find a way of controlling the cystatin fibrillization process, the interactions between monoclonal antibody Cyst-13 and cystatin C were studied in detail. The present work describes the determination of the epitope of hCC to a monoclonal antibody raised against cystatin C, Cyst-13, by MALDI mass spectrometry, using proteolytic excision of the immune complex. The shortest epitope sequence was determined as hCC(107-114). Affinity studies of synthetic peptides revealed that the octapeptide with epitope sequence does not have binding ability to Cyst-13, whereas its longer counterpart, hCC(105–114), binds the studied antibody. The secondary structure of the peptides with epitope sequence was studied using circular dichroism and NMR spectroscopy

Interaction of serum amyloid A with human cystatin Cuidentification of binding sites

Serum amyloid A (SAA) is a multifunctional acute-phase protein whose natural role seems to be participation in many physiologic and pathological processes. Prolonged increased SAA level in a number of chronic inflammatory and neoplastic diseases gives rise to reactive systemic amyloid A amyloidosis, where the N-terminal 76-amino acid residue-long segment of SAA is deposited as amyloid fibrils. Recently, a specific interaction between SAA and the ubiquitous inhibitor of cysteine proteaseshuman cystatin C (hCC)has been described. Here, we report further evidence corroborating this interaction, and the identification of the SAA and hCC binding sites in the SAAhCC complex, using a combination of selective proteolytic excision and high-resolution mass spectrometry. The shortest binding site in the SAA sequence was determined as SAA(86104), whereas the binding site in hCC sequence was identified as hCC(96102). Binding specificities of both interacting sequences were ascertained by affinity experiments (ELISA) and by registration of mass spectrum of SAAhCC complex. Copyright (c) 2012 John Wiley & Sons, Ltd

Cysteine cathepsins and their inhibitors in supernatants of CF sputum.

<p>Only representative samples are shown. (A) Proteins (30 µg/well) were separated by 15% SDS-PAGE under reducing conditions, transferred to nitrocellulose membranes, and analyzed with polyclonal antibodies against human cathepsins B, H, L and S. (+): <i>Pseudomonas aeruginosa</i>-colonized CF sputum; (−): <i>Pseudomonas aeruginosa</i>-negative CF sputum.◃, single-chain cathepsin B; ◂, double-chain cathepsin B; ←, mature cathepsins S and H; ★, proforms. (B) Immunostaining with polyclonal anti-cystatin C antibody and anti-kininogen antibody. (+): <i>Pseudomonas aeruginosa</i>-colonized CF sputum; (−): <i>Pseudomonas aeruginosa</i>-negative CF sputum. Control: Cyst, Cystatin C; HK, HMWK. Recombinant human cystatin C (R&D systems) has an additional C-terminal 10 His-tag and an apparent molecular mass of 17 kDa, according to the supplier. (C) Supernatants of CF sputum incubated with Biot-LVG-CHN₂ (30 µM), for 1 h at 37°C <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025577#pone.0025577-Florent1" target="_blank">[25]</a>. Other samples were pre-incubated with E-64, CA-074, and Mu-Leu-Hph-VSPh prior to adding the biotinylated activity-based probe. Samples were separated by 12% SDS-PAGE, electroblotted and incubated with extravidin-peroxidase conjugate. The peroxidase activity was revealed by chemiluminescence. WB: individual cathepsins B, H, L and S immunoblotted as control. Control: (−), no pre-incubation with E-64; (+), pre-incubation with E-64 prior to adding Biot-LVG-CHN2. Sputum: E-64, pre-incubation with E-64; CA, pre-incubation with CA-074; VS, pre-incubation with Mu-Leu-HphVSPh.</p