Investigation of relationship between musculoskeletal disorders and working conditions among workers at a pharmaceutical industry in Iran (2011-2012)

Musculoskeletal disorders may be observed in all industries and professions and most of these disorders are related to the back, upper and lower extremities of the body organs. In Pharmaceutical industry, almost lack of standard ergonomic conditions and sometimes can cause outbreaks of diseases and musculoskeletal disorders in various parts of the body. The aim of this study was to determine the relationship between musculoskeletal disorders and working on the packaging section of the pharmaceutical industry. The Nordic questionnaire and Rula method were used for collection of data and 392 workers were selected as the subjects of study. Based on the results of this study, (28.5%) of workers working in Packaging Unit complained of severe pain and discomfort in their neck, (23.7%) in their shoulder, (27.9%) in their hand and wrist and (33.2%) complained of severe pain in their back. The results indicate that workers in this industry could show musculoskeletal disorders based on age, education, gender and working conductions

The tyrosine kinase receptor ROR1 is constitutively phosphorylated in chronic lymphocytic leukemia (CLL) cells.

Phosphorylation of receptor tyrosine kinases (RTKs) has a key role in cellular functions contributing to the malignant phenotype of tumor cells. We and others have previously demonstrated that RTK ROR1 is overexpressed in chronic lymphocytic leukemia (CLL). Silencing siRNA downregulated ROR1 and induced apoptosis of CLL cells. In the present study we analysed ROR1 isoforms and the phosphorylation pattern in CLL cells (n=38) applying western blot and flow-cytometry using anti-ROR1 antibodies and an anti-phospho-ROR1 antibody against the TK domain. Two major ROR1 bands with the size of 105 and 130 kDa respectively were identified, presumably representing unglycosylated (immature) and glycosylated (mature) ROR1 respectively as well as a 260 kDa band which may represent dimerized ROR1. A ROR1 band of 64 kDa that may correspond to a C-terminal fragment was also noted, present only in the nucleus. The 105 kDa ROR1 isoform was more frequently expressed in non-progressive as compared to progressive CLL patients (p=0.03). The 64, 105, 130 and 260 kDa bands were constitutively phosphorylated both at tyrosine and serine residues. Phosphorylation intensity of the mature (130 kDa) isoform was significantly higher in progressive than in non-progressive disease (p<0.001). Incubation of CLL cells with a mouse anti-ROR1 KNG or an anti-ROR1 CRD mAb respectively induced dephosphorylation of ROR1 before entering apoptosis. In conclusion CLL cells expressed different isoforms of ROR1 which were constitutively phosphorylated. The mature, phosphorylated ROR1 isoform was associated with a progressive disease stage. Targeting ROR1 by mAbs induced specific dephosphorylation and leukemic cell death. ROR1 might be an interesting therapeutic target

Anti-ROR1 mAbs induced dephosphorylation of ROR1 and subsequent apoptosis of CLL cells.

<p>Representative experiments of time-dependent ROR1 dephosphorylation and apoptosis in vitro incubated with an anti-KNG (4A7) (A and C) and anti-CRD (1D8) (B and D) anti-ROR1 mAb respectively. Representative experiments from two progressive (CLL 5102 and CLL 5116) CLL patients are shown. Leukemic cells were incubated for 20 min, 1, 4 hours with a non-relevant IgG1 isotype control mAb (-) and the ROR1 mAbs (+). Cells were harvested and lysed for western blot experiments (A and B). The non-relevant isotype control mAb did not induce dephosphorylation of ROR1 (130 kDa) while the anti-ROR1 mAb induced dephosphorylation already after 20 min, which increased by time. The intensity of pROR1 was measured by ImageJ software. The ratio of pROR1/ROR1 intensity of treated sample to pROR1/ROR1 intensity of untreated sample (relative intensity) is shown in a histogram. A value <1 indicates dephosphorylation of ROR1 after treatment with the anti-ROR1 mAb compared to the non-relevant isotype control mAb. Apoptosis of CLL cells (Annexin V⁺/PI⁺) after 0, 12 and 18 h incubation with the anti ROR1 mAbs without the addition of immune effector cells or complement (C and D). An anti-CD20 mAb (ofatumumab) did not induce dephosphorylation of ROR1 after 4h incubation in two CLL patients. A representative experiment is shown for one of the CLL patients (E).</p

ROR1 isoforms and phosphorylation in CLL patients.

<p>Representative experiments using non-immunoprecipitated CLL cell lysates from 5 CLL patients showing phosphorylated dimerized ROR1 (260 kDa), fully glycosylated (130 kDa) and non-glycosylated (105 kDa) ROR1 molecules (A). NP indicates non-progressive disease and P progressive disease. As controls, CLL cell lysates immunoprecipitated with a non-relevant mAb (mouse IgG1 isotype) were used. No bands could be seen. Furthermore, in PBMC of healthy donors, no bands were detected (data not shown). Representative experiments of PBMC using surface staining for ROR1 (left panel) and intracytoplasmic staining for pROR1 (right panel) in three CLL patients (B). Expression of ROR1 isoforms in the cytoplasm (C), nucleus (N) and total cell lysate (TCL) of leukemic CLL cells (C). Confirmation of protein localization was done using antibodies against α/β –tubulin, histone H3 and nucleolin before analysing the expression pattern of ROR1. </p

Differential phosphorylation of ROR1 isoforms in CLL in relation to disease activity.

<p>pROR1/ROR1 intensity (mean ± SEM) of the 105, 130 and combined 105+130 kDa bands in progressive (■) (n=21) and non-progressive (☐) CLL patients (n=17). *** p= 0.001.</p