Working conditions of resident SHOs.
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Biomedical subjects
Publications and source records attributed to S Porter.
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The regulatory protein NIFA activates transcription of nitrogen fixation (nif) operons by the sigma 54 holoenzyme form of RNA polymerase. NIFA from Klebsiella pneumoniae activates transcription from the nifH promoter in vitro; in addition, the integration host factor, IHF, binds between the nifH promoter and an upstream binding site for NIFA. We demonstrate here that IHF greatly stimulates NIFA-mediated activation of nifH transcription in vitro and thus that the two factors are functionally synergistic. Electron micrographs indicate that IHF bends the DNA in the nifH promoter regulatory region. Although IHF binds close to the nifH promoter, it does not directly stimulate binding of sigma 54 holoenzyme. Rather, the IHF-induced bend may facilitate productive contacts between NIFA and sigma 54 holoenzyme that lead to the formation of open complexes. IHF binds to nif promoter regulatory regions from a variety of organisms within the phylum "purple bacteria," suggesting a general ability to stimulate NIFA-mediated activation of nif transcription.
Female Sprague-Dawley rats (n = 72), 6 months old, underwent either sham operation, oophorectomy, adrenalectomy, or combined oophorectomy and adrenalectomy (O&A). They were all maintained on normal saline ad libitum and 20 g/day 1.1% calcium chow. Nine weeks after operation, the trabecular bone volume of the distal femoral shaft was significantly lower (P less than 0.001) in the adrenalectomized (11.1%), oophorectomized (7.0%), and O&A (8.3%) animals than in sham-operated animals (19.8%). Eighteen weeks after operation, the trabecular bone volume in O&A animals had fallen to a mean of 3.8% (sham 17.0%), and the length of the femur had increased to 38.8 mm after O&A (sham 36.8 mm, P less than 0.01). O&A animals treated with 0.35 mg/kg/week nandrolone decanoate from 9 weeks postoperatively onward, had twice the femoral trabecular bone volume of untreated animals at 18 weeks (P less than 0.05). By contrast, no significant differences were found in vertebral body trabecular bone between any groups, including groups receiving treatment with androgens. We have found that, by 9 weeks after operation, adrenalectomy alone causes significant loss of metaphyseal trabecular bone, similar to the progressive loss seen after oophorectomy.
Occupational injuries to dental hospital personnel over 9 years (1980-1988 inclusive) were studied. Some 300 accidents were reported. Over one-half were sharps injuries. Over one-quarter were falls or collisions. Eye injuries accounted for some 10 per cent of incidents, burns and scalds the remainder. Serious complications were rare, but many of the injuries could have been avoided.
The NtrC protein activates transcription of the glnA operon of enteric bacteria by stimulating the formation of stable "open" complexes by RNA polymerase (sigma 54-holoenzyme form). To regulate the glnA promoter, NtrC binds to sites that have the properties of transcriptional enhancers: the sites will function far from the promoter and in an orientation-independent fashion. To investigate the mechanism of enhancer function, we have used electron microscopy to visualize the interactions of purified NtrC and RNA polymerase with their DNA binding sites and with each other. Under conditions that allow the formation of open complexes, about 30% of DNA molecules carry both RNA polymerase and NtrC bound to their specific sites. Of these, about 15% form looped structures in which NtrC and the RNA polymerase-promoter complex are in contact. The length of the looped DNA is that predicted from the spacing that was engineered between the enhancer and the glnA promoter (390 base pairs). As expected for activation intermediates, the looped structures disappear when RNA polymerase is allowed to transcribe the DNA. We conclude that the NtrC enhancer functions by means of a direct association between DNA-bound NtrC and RNA polymerase (DNA-looping model). Association of DNA-bound proteins appears to be the major mechanism by which different types of site-specific DNA transactions are localized and controlled.
It took two years to transform the oncology unit at Mount Carmel Medical Center, Columbus, OH, from a cold, sterile-looking environment into a comfortable, homey area. Now bright quilts adorn the walls, carpeting softens the floors in hallways and "family areas," flowered curtains balloon at windows, and a kitchen is available so visitors can fix favorite foods for patients and pour themselves a cup of coffee. The family of the late Jane Barks Ross, a former cancer patient at the hospital, provided a good chunk of the money for the project, and the hospital auxiliary chipped in additional funds. A design firm was hired to consult with staff and patient and come up with a scheme that everyone liked hospital construction workers did most of the labor, aided by off-duty staff and auxiliary members while the patients themselves had front-row seats to watch the progress.
The normal neuromuscular synapse is formed by the intimate association of nerve endings, postsynaptic end-plate foldings in the muscle fiber, and nonmyelinating Schwann cells (SC) sealing the synaptic ramifications. Because SC have been recognized recently to have an immunogenic potential inducible to present protein autoantigens to autoimmune T lymphocytes, and considering their close proximity to the acetylcholine receptor (AChR)-bearing postsynaptic membranes, presentation of soluble and membrane vesicle-bound AChR to appropriate T cells was investigated. Short-term monolayer cultures of SC isolated from neonatal rat sciatic nerves, as well as cells of an immortalized SC line of similar origin, were fully able to present the relevant molecular epitopes to major histocompatibility complex (MHC) compatible AChR-specific T line lymphocytes immunogenically. Presentation of AChR was restricted by RT1.B (I-A) MHC class II products. Both types of cultured rat SC were inducible to expression of MHC class I and II products, and they were able to phagocytose AChR-enriched membrane vesicles preferentially. In contrast, phagocytosis of latex particles by SC was negligible. These data qualify perisynaptic SC as potential presenter cells of autoimmunogenic AChR in myasthenia gravis. Thus, SC may play a critical and as-yet unpredicted regulatory role in the cellular pathogenesis of myasthenia gravis.
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Recurrent aphthous stomatitis (RAS) is a common oral disorder, however, despite detailed clinical, immunologic, hematologic and microbiologic investigation, the etiology of RAS remains unknown. At present, topical steroids and antimicrobial mouth rinses are the mainstays of treatment, but there is still no means of preventing recurrence of the oral ulceration.
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We have recently reported the immortalization of primary Schwann cells isolated from sciatic nerves of normal neonatal rats. The cells were maintained under continuous mitogenic stimulation with glial growth factor and forskolin, achieving immortalization after 12 to 15 weeks without the use of viral infection, oncogene transformation or chemical carcinogens. The immortalized cells (1.17 cells) initially retain the capability to recognize and attach to peripheral neurons in culture as well as the ability to myelinate those neurons. The functional capacity of the cells gradually diminishes in culture, such that late passage cells can ensheath neurons but cannot form a myelin sheath. Both normal and immortalized cells secrete comparable amounts of autocrine growth factor activity in culture that can be regulated by extracellular matrix proteins. The difference between quiescent and immortalized Schwann cells seems to lie not in the production of growth factor but rather in the relative ability to respond to the factor(s). To test the potential of the immortalized Schwann cells for the ability to form tumours in vivo, we injected equal numbers of primary or immortalized Schwann cells into the sciatic nerve of adult syngenic rats and allowed them to incubate there for 6 to 13 weeks, whereupon the injected nerves were inspected for tumour formation. In every case (N = 3) the primary cells had no effect whereas every injection of immortalized cells (N = 5) resulted in a solid cellular mass surrounding the injected nerve. The tumours were encapsulated masses of actively dividing Schwann-like cells that surrounded but did not invade the nerve fascicle.(ABSTRACT TRUNCATED AT 250 WORDS)
A commercially produced domiciliary fetal monitoring (DFM) system was assessed in four centres in the UK (Bristol, Glasgow, London and Nottingham) chosen to allow for comprehensive assessment in various settings in many different women. Overall, 825 recordings were made from 368 women (2.24 per woman). There were 56 unsuccessful attempts at transmission (6.8%), most were due to problems with telephone equipment. The system worked most efficiently when a dedicated direct line was used. The data transmission time varied between 40 and 60 s. The median telephone time (including data transmission and conversation) with a dedicated direct line was 7 min. Mean acceptance times from the four centres were between 70 and 80%. All recordings with acceptance times of 40% or more were interpretable. Ten recordings were abnormal. The women and mid-wives were equally proficient at using the DFM system. The DFM system represents an important addition to current methods of fetal assessment. Specific guidelines are outlined.
Expression of antibody heavy- and light-chain genes by transfection permits the production of monoclonal antibodies with improved biological and antigen-binding properties. The immunoglobulin genes are placed in vectors containing a gene for encoding a protein that provides a biochemically selectable function in eukaryotic cells; these vectors are transfected into myeloma and hybridoma cells. Selection of drug-resistant cells permits the efficient isolation of the rare cells that express the transfected DNA. By placing heavy and light chains on plasmids with different selectable markers, one can deliver heavy- and light-chain genes simultaneously to the same cell. The transfected immunoglobulin genes are efficiently expressed and the proteins produced are a faithful mirror of the genes that were introduced. Using the standard techniques of genetic engineering and gene transfection, we can now produce antibodies of widely varying structures, including chimeric antibodies with segments derived from different species. These antibodies provide useful reagents to study structure-function relationships within the antibody molecule. Ultimately it will be possible to produce a new generation of antibody molecules with improved antigen-binding properties and effector functions.
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Schwann cells derived from neonatal rat sciatic nerve are quiescent in culture unless treated with specific mitogens. The use of glial growth factor (GGF) and forskolin has been found to be an effective method for stimulating proliferation of Schwann cells on a poly(L-lysine) substratum while maintaining their ability to myelinate axons in vitro. We find that repetitive passaging of Schwann cells with GGF and forskolin results in the loss of normal growth control; the cells are able to proliferate without added mitogens. The immortalized cells grow continuously in the absence of added growth factor and in the presence or absence of serum yet continue to express distinctive Schwann cell-surface antigens. The cells can associate with axons in culture, deposit a basal lamina, and ensheath axons, but they gradually lose their capacity to myelinate axons. The immortalized cells release growth-promoting activity into their culture medium. The released activity is effective in stimulating proliferation of primary Schwann cells that retain normal growth properties. Extracellular matrix molecules (laminin and fibronectin) augment the response of primary Schwann cells to the secreted mitogen. Quiescent primary Schwann cells also secrete a growth factor into their culture medium, but its activity is detectable only in the presence of added laminin or fibronectin. The results suggest that both normal and immortalized Schwann cells secrete an autocrine growth factor. Response to the autocrine factor appears to entail a multicomponent mechanism. Unlike primary cells, immortalized Schwann cells have the capacity to secrete all of the necessary components and to respond to them constitutively.
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