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Cloning and expression of a human kinesin heavy chain gene: interaction of the COOH-terminal domain with cytoplasmic microtubules in transfected CV-1 cells.
To understand the interactions between the microtubule-based motor protein kinesin and intracellular components, we have expressed the kinesin heavy chain and its different domains in CV-1 monkey kidney epithelial cells and examined their distributions by immunofluorescence microscopy. For this study, we cloned and sequenced cDNAs encoding a kinesin heavy chain from a human placental library. The human kinesin heavy chain exhibits a high level of sequence identity to the previously cloned invertebrate kinesin heavy chains; homologies between the COOH-terminal domain of human and invertebrate kinesins and the nonmotor domain of the Aspergillus kinesin-like protein bimC were also found. The gene encoding the human kinesin heavy chain also contains a small upstream open reading frame in a G-C rich 5' untranslated region, features that are associated with translational regulation in certain mRNAs. After transient expression in CV-1 cells, the kinesin heavy chain showed both a diffuse distribution and a filamentous staining pattern that coaligned with microtubules but not vimentin intermediate filaments. Altering the number and distribution of microtubules with taxol or nocodazole produced corresponding changes in the localization of the expressed kinesin heavy chain. The expressed NH2-terminal motor and the COOH-terminal tail domains, but not the alpha-helical coiled coil rod domain, also colocalized with microtubules. The finding that both the kinesin motor and tail domains can interact with cytoplasmic microtubules raises the possibility that kinesin could crossbridge and induce sliding between microtubules under certain circumstances
Should Any Workplace Be Exempt from Smoke-Free Law: The Irish Experience
Background. In 2004, the Irish Government introduced national legislation banning smoking in workplaces; with exemptions for “a place of residence”. This paper summarises three Irish studies of exempted premises; prisons, psychiatric hospitals and nursing homes. Methods. PM2.5 and nicotine were measured in nursing homes and psychiatric hospitals, in addition to ultrafine particles in the hospitals. In the prisons, officers (n = 30) completed exhaled breath Carbon Monoxide (CO) measurements. Questionnaires determined officers' opinion on introducing smoking prohibitions in prisons. Nursing home smoking policies were examined and questionnaires completed by staff regarding workplace secondhand smoke (SHS) exposure. Findings. Ultrafine particle concentrations in psychiatric hospitals averaged 130,000 cm3, approximately 45% higher than Dublin pub (35.5 μg/m3) pre ban. PM2.5 levels in psychiatric hospitals (39.5 μg/m3) were similar to Dublin pubs (35.5 μg/m3) pre ban. In nursing homes permitting smoking, similar PM2.5 levels (33 μg/m3) were measured, with nicotine levels (0.57 μg/m3) four times higher than “non-smoking” nursing homes (0.13 μg/m3). In prisons, 44% of non-smoking officers exhibited exhaled breath CO criteria for light to heavy smokers. Conclusions. With SHS exposure levels in some exempted workplaces similar to Dublin pubs levels pre ban, policies ensuring full protection must be developed and implemented as a right for workers, inmates and patients
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