Nucleo-cytoplasmic localization of influenza virus nucleoprotein depends on cell density and phosphorylation

Influenza virus nucleoprotein (NP) plays a major role in the nucleus during virus replication, and is a mediator of viral ribonucleoprotein nuclear import during entry. NP is localized primarily in the nucleus, but can undergo nucleo-cytoplasmic shuttling in heterokaryons (Whittaker et al., 1996a. J. Virol. 70, p. 2743). We have studied NP localization using a stable cell line (3PNP-4) that expresses NP. Intracellular localization of NP was markedly affected by the density of the cell monolayer. It was nuclear in cells grown in sparse culture, but cytoplasmic in dense culture. In phorbol ester-stimulated cells NP was cytoplasmic, but relocalized to the nucleus after treatment with a protein kinase inhibitor. Cell density and phosphorylation-dependent localization of NP appeared to be independent of cell type. Our data suggest that a phosphorylation event is needed either for nuclear export, or to regulate retention of NP in the nucleus, and that regulation may be mediated by kinases activated by cell-cell contact.     

Development of abnormal tissue architecture in transplanted neonatal rat myocytes

BACKGROUND: Most myocardial cell transplant studies focus on demonstration of improved function; however, such improvement depends on the development of appropriate tissue structure. Thus, our aim was to assess the architectural changes that occurred after cell transplant into normal and infarcted myocardium. METHODS: Male neonatal cells (1 to 2 days old) were injected into the left ventricular free wall of adult female rats. The tissue was examined 0 to 1 days and 1 to 2, 4 to 6, and 12 weeks later in noninfarcted hearts and 6 months after transplant into infarcts. In histologic sections, we assessed the cells' retardation of polarized light (to measure development of contractile elements), two-dimensional cell orientation, cell nuclear morphology, and collagen content. RESULTS: The transplant cells' retardation of polarized light gradually increased to 81% of that of host cells after 6 months (p < 0.001). The transplant cells were disorganized and although their nuclei increased in size, they always had a rounded appearance. Collagen content in the transplant was 210% to 430% higher than in host tissue (p < 0.01). In addition, scar collagen always separated transplant and host cells. CONCLUSIONS: One architectural feature, the rounded nuclei, provided a distinctive marker to identify transplanted cells. Nevertheless, the transplants' inhibited muscle development together with disorganization, separation from the host muscle, and a substantial increase in collagen resulted in a structure unlikely to play an active role in systolic function.     

Virus nuclear import

Many viruses replicate in the nucleus of their host cells. To gain access to this compartment, they must navigate their way from the cell surface, through the endosomal or plasma membrane, across a crowded cytoplasm and finally cross the nuclear envelope. Entry into the nucleus typically occurs via nuclear pores, which have a strict size limit that most viruses cannot traverse without prior uncoating or disassembly. While the majority of the viruses make use of the cellular nuclear import machinery for their nuclear transport, a growing number of viruses seem to use specialized, virus-encoded routes of nuclear import. For nuclear-replicating viruses, entry into the nucleus is highly dependent on prior trafficking and uncoating events that act to prime the virus genome for its final destination.     

Lead exposure in radiator repair workers: a survey of Washington State radiator repair shops and review of occupational lead exposure registry data

Radiator repair workers in Washington State have the greatest number of very elevated (> or =60 microg/dL) blood lead levels of any other worker population. The goals of this study were to determine the number of radiator repair workers potentially exposed to lead; estimate the extent of blood lead data underreporting to the Occupational Lead Exposure Registry; describe current safety and health practices in radiator repair shops; and determine appropriate intervention strategies to reduce exposure and increase employer and worker awareness. Lead exposure in Washington State's radiator repair workers was assessed by reviewing Registry data and conducting a statewide survey of radiator repair businesses. This study revealed that a total of 226 workers in Washington State (including owner-operators and all employees) conduct repair activities that could potentially result in excessive exposures to lead. Approximately 26% of radiator repair workers with elevated blood lead levels (> or =25 microg/dL) were determined to report to Washington State's Registry. This study also revealed a lack of awareness of lead's health effects, appropriate industrial hygiene controls, and the requirements of the Lead Standard. Survey respondents requested information on a variety of workplace health and safety issues and waste management; 80% requested a confidential, free-of-charge consultation. Combining data derived from an occupational health surveillance system and a statewide mail survey proved effective at characterizing lead exposures and directing public health intervention in Washington State.     

Allelotyping in mycosis fungoides and Sézary syndrome: common regions of allelic loss identified on 9p, 10q, and 17p

Allelotyping studies have been extensively used in a wide variety of malignancies to define chromosomal regions of allelic loss and sites of putative tumor suppressor genes; however, until now this technique has not been used in cutaneous lymphoma. We have analyzed 51 samples from patients with mycosis fungoides and 15 with Sézary syndrome using methods to detect loss of heterozygosity. Micro satellite markers were selected on 15 chromosomal arms because of their proximity to either known tumor suppressor genes or chromosomal abnormalities identified in previous cytogenetic studies in cutaneous lymphoma. Allelic loss was present in 45% of patients with mycosis fungoides and 67% with Sézary syndrome. Loss of heterozygosity was found in over 10% of patients with mycosis fungoides on 9p, 10q, 1p, and 17p and was present in 37% with early stage (T1 and T2) and 57% with advanced disease (T3 and T4). Allelic loss on 1p and 9p were found in all stages of mycosis fungoides, whereas losses on 17p and 10q were limited to advanced disease. In Sézary syndrome high rates of loss of heterozygosity were detected on 9p (46%) and 17p (42%) with lower rates on 2p (12%), 6q (7%), and 10q (12%). There was no significant difference in the age at diagnosis or number of treatments received by those with loss of heterozygosity and those without, suggesting that increasing age and multiple treatments do not predispose to allelic loss. These results provide the basis for further studies defining more accurately chromosomal regions of deletions and candidate tumor suppressor genes involved in mycosis fungoides and Sézary syndrome.