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Biomedical subjects

T Kelly

Publications and source records attributed to T Kelly.

At least 91 records · Page 5Linked to original sources
PubMed1988

Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.

The replication of simian virus 40 (SV40) DNA is largely dependent upon cellular replication proteins. To define these proteins we have made use of a cell-free system that is capable of replicating plasmid DNA molecules containing the SV40 origin of replication. Systematic fractionation-reconstitution experiments indicate that there are a minimum of six cellular proteins that are required for efficient viral DNA replication in vitro. We report here the purification of one of these proteins, replication protein A (RP-A), to homogeneity. RP-A is a multisubunit protein that contains four tightly associated polypeptides of 70, 53, 32, and 14 kDa. Partial proteolysis experiments indicate that the 53-kDa polypeptide is closely related to the 70-kDa polypeptide, suggesting that it may be a proteolytic fragment of the larger subunit. RP-A is absolutely required for reconstitution of SV40 DNA replication in vitro. The purified protein binds to single-stranded DNA and is required for the large tumor (T)-antigen-mediated unwinding of DNA molecules containing the SV40 origin of DNA replication. These properties are consistent with the possibility that RP-A plays a central role in the generation of a single-stranded region at the origin prior to initiation of DNA synthesis. The protein may also function to facilitate unwinding of the parental DNA strands during the elongation phase of SV40 DNA replication.

Antigens, Viral, Tumor↗
PubMed1988

Time of exposure and risk of HIV infection in homosexual partners of men with AIDS.

We interviewed, and tested for HIV antibody, 117 homosexual men who had been regular sexual partners of men who developed acquired immunodeficiency syndrome (AIDS); 85 tested seropositive. Receptive anal intercourse with the index AIDS case and number of different sexual partners with whom subjects were anally receptive were both risk factors. Controlling for the number of partners with whom subjects were anally receptive, we found that the odds ratio of receptive anal intercourse with the case was infinite (95% confidence intervals, 3.3-infinity) if sexual contact continued up to or beyond the time of diagnosis, while the odds ratio was 1.0 (95% CI 0.3-3.2) if contact ceased before the case's AIDS diagnosis. Risk was not associated with the duration or frequency of contact. Our data suggest that the potential for sexual transmission from an HIV-infected person may be greater close to or after the onset of disease.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Upper gastrointestinal endoscopy after cardiac transplantation.

Three hundred four patients have undergone cardiac transplantation at the University of Pittsburgh since 1980. Twenty patients have required 27 upper gastrointestinal (GI) endoscopic procedures. After heart transplantation the primary indications for endoscopy were epigastric pain (six patients with gastritis, one with multiple shallow gastric ulcers, and one with normal test results), mild upper GI bleeding (four patients with esophagitis, two with gastritis, and two with multiple shallow gastric ulcers), dysphagia and odynophagia (two patients with esophagitis), persistent nausea and vomiting (one with normal test results), lower GI bleeding (one with normal test results), and routine follow-up (one with normal test results). After heart-lung transplantation the primary indications for the endoscopy were massive upper GI bleeding (three patients with actively bleeding duodenal ulcers), dysphagia and odynophagia (one patient with esophagitis), mild upper GI bleeding (one patient with gastritis), and routine follow-up (one patient with normal test results). No complications resulted from endoscopy. The procedures were performed in the GI suite without cardiac monitoring. Prophylactic antibiotics were not routinely administered. No patient had a fungal infection of the upper GI tract--a finding attributed to the prophylactic use of nystatin in all patients. Opportunistic viral infections were identified histologically in six patients, including two patients with actively bleeding duodenal ulcers. The possibility of opportunistic viral infections in this immunosuppressed group required aggressive diagnostic techniques, including endoscopy and biopsy, which can be safely performed after cardiac transplantation.

Adult↗
PubMed1987

Purification of two smooth muscle glycoproteins related to integrin. Distribution in cultured chicken embryo fibroblasts.

We have purified two membrane glycoproteins from chicken gizzard smooth muscle. In the presence of reducing agents, these proteins have molecular weights on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 165,000 and 130,000, but they migrate at 165,000 and 110,000 without reduction. The two proteins can also be isolated as a complex in buffers containing physiologic salt concentrations. This complex has physical properties similar to two proteins of the integrin family of receptors for extracellular matrix proteins, the cell substratum attachment antigen from chicken embryos, and the glycoprotein IIb IIIa complex from mammalian platelets. When the smooth muscle complex is visualized by electron microscopy, it has a striking resemblance to both avian integrin and the glycoprotein IIb IIIa complex. Smooth muscle is a good source of the 165,000 and 130,000 proteins, and purification of both the individual subunits and the complex is achieved using conventional biochemical techniques. Antibodies directed against the 130,000 protein cross-react with integrin but do not cross-react with the 165,000 protein. Immunofluorescence microscopy using these antibodies reveals staining of fibroblast focal contacts and fibrillar streaks which coalign with fibronectin. Whereas monoclonal antibodies against integrin label the periphery of the focal contact more intensely than the center, the anti-130,000-protein serum stains the entire focal contact. Antibodies directed against the 165,000 protein also stain focal contacts and fibrillar streaks of fibroblasts in tissue culture. On the basis of similar physical properties, biochemical characteristics, and immunological cross-reactivity we conclude that the 165,000/130,000 complex is a smooth muscle integrin.

Animals↗
PubMed1987

Comparison of RapID-ANA and Minitek with a conventional method for biochemical identification of anaerobes.

Two micromethods for the identification of anaerobes, one requiring growth (Minitek) and one nongrowth dependent (RapID-ANA), were compared with a conventional identification culture system. For 222 clinical isolates, RapID-ANA agreed with PRAS in 187 (84%) and Minitek agreed for only 170 strains (76%). Both systems identified common isolates well, but encountered some difficulty in identifying less common clostridia and Gram-negative bacilli. Although adequate for most strains, the results from both systems should be interpreted with caution, particularly for less frequently isolated species.

Bacteria, Anaerobic↗
PubMed1987

The role of the community psychiatric nurse in improving treatment compliance in alcoholics.

Patient compliance in alcoholism treatment remains an under-researched topic. Some factors as to why patients comply to treatment have been studied but the intervention of nursing staff who work in the community has not been extensively researched. This study looks at the effect, if any, of having community psychiatric nurses following up a randomly-selected group of new patients referred to the Withington Hospital Alcoholism Treatment Unit. This study looks at the involvement of community psychiatric nurses in their initial treatment.

Alcoholism↗
PubMed1987

Adhesion plaques: sites of transmembrane interaction between the extracellular matrix and the actin cytoskeleton.

In this paper we review what is known about the organization of adhesion plaques, the regions where cells in culture adhere most tightly to the underlying substratum. These specialized areas of the plasma membrane serve as attachment sites for stress fibres. A major objective has been to determine how microfilament bundles are anchored at such regions. In their morphology and composition adhesion plaques resemble the adhesions fibroblasts make to the extracellular matrix. Some extracellular matrix components have been identified on the outside face of adhesion plaques. Within the plasma membrane of adhesion plaques, extracellular matrix receptors, such as the fibronectin receptor (integrin), have been identified. This transmembrane glycoprotein complex has been shown to bind the cytoplasmic protein talin, which, in turn, associates with vinculin. These proteins establish a transmembrane chain of attachment between the extracellular matrix and the cytoskeleton, although how the actin filaments interact with these components remains to be determined. Besides having a structural function, adhesion plaques may also be regions where regulatory signals are transmitted across the membrane. Consistent with this idea has been the finding that various tyrosine kinases and a calcium-dependent protease are concentrated at the cytoplasmic aspect of adhesion plaques. Furthermore, several adhesion plaque proteins become phosphorylated during cell transformation by Rous sarcoma virus. In future work it will be important to determine how such modifications affect the interactions of these proteins and the stability of adhesion plaques.

Actins↗
PubMed1986

Functional organization of the simian virus 40 origin of DNA replication.

To define the sequence elements involved in initiation of DNA synthesis at the simian virus 40 origin of replication, we determined the relative replication efficiencies in vitro and in vivo of templates containing a variety of mutations within the origin region. Replication of the mutants in vitro was assayed by the cell-free DNA replication system that we recently described (J.J. Li and T.J. Kelly, Proc. Natl. Acad. Sci. USA 81:6973-6977, 1984; J.J. Li and T.J. Kelly, Mol. Cell. Biol. 5:1238-1246, 1985), and replication in vivo was assayed after transfection of the mutant templates into COS-1 cells. The minimal origin of replication defined by both assays included a 15-base-pair (bp) imperfect inverted repeat, a 27-bp perfect inverted repeat, and a 17-bp A/T-rich region. T-antigen binding site I was not required for DNA replication, but its presence increased replication efficiency severalfold both in vitro and in vivo. Although SP1 binding sites and enhancers had little or no effect on replication in vitro, the presence of either element markedly increased replication in vivo. Thus, the biological role of these elements is not restricted to stimulating transcription but may be more general.

Antigens, Polyomavirus Transforming↗
PubMed1984

Antibiotic prophylaxis in high-risk biliary operations: multicenter trial of single preoperative ceftriaxone versus multidose cefazolin.

A prospective, double-blind trial of a single preoperative dose of ceftriaxone, a new long-acting cephalosporin, versus one preoperative and three postoperative doses of cefazolin was carried out in 81 patients at high risk of infection after biliary surgery. Indications for antibiotic prophylaxis included recent or ongoing cholecystitis (52 patients), common duct stones (14 patients), common duct obstruction (3 patients), and age greater than 70 years (22 patients). Intraoperative bile cultures were positive in 7 of 41 patients (17.1 percent) given ceftriaxone and 12 of 40 patients (30 percent) given cefazolin, but there were no wound infections in either group. Neither regimen was associated with significant antibiotic resistance. Side effects, such as proteinuria and elevated liver transaminases and alkaline phosphatase levels, were transient and not definitely related to the antibiotics. We conclude that a single preoperative dose of ceftriaxone is as effective as multiple perioperative doses of cefazolin in the prophylaxis of infection associated with biliary tract surgery.

Adolescent↗
PubMed1984

Abnormality of cartilage collagen in a patient with unclassified chondrodystrophy.

We have described a previously unrecognized chondrodystrophy characterized by short-limbed dwarfism, blue sclera, severe cardiopulmonary problems, and failure of postnatal growth. The first of two siblings thus affected died at age 6 months following attempted correction of an atrial septal defect. Growth plate cartilage from multiple sites obtained at autopsy showed a marked abnormality of architecture on the light microscopic level. Biochemical studies demonstrated an absence of normal alpha 1(II) collagen in costochondral junction growth plate cartilage and an appearance of the major collagen in a band which comigrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with 3 alpha collagen. Cartilage extracted from structural rib appeared to be normal.

Cartilage↗
PubMed1983

Isodicentric X chromosome in a girl with gonadal dysgenesis.

Chromosomal analysis on a 15-year-old girl with amenorrhea and lack of secondary sexual development showed a normal X chromosome and an isodicentric X chromosome, 46,X, idic(x) (pter leads to q23::q23 leads to pter). Thus the patient has duplicated and deleted X chromosome material. There was no evidence of 45,X mosaicism. Brd U-labeling revealed the abnormal X to be late-replicating.

Adolescent↗
PubMed1982

Proteins involved in the attachment of actin to the plasma membrane.

Proteins that may be involved in two types of actin-membrane association are discussed. The first set includes alpha-actinin, vinculin, fimbrin and a new cytoskeletal protein that are all concentrated in adhesion plaques, those regions of cultured fibroblasts where bundles of actin microfilaments terminate and where the plasma membrane comes close to the underlying substrate. The properties of non-muscle alpha-actinin suggest that it functions to cross-link actin filaments and thereby stabilize microfilament bundles rather than functioning in their attachment to the membrane. Fimbrin also appears to be involved in bundling of filaments rather than in attachment. In contrast, vinculin binds to the ends of actin filaments in vitro and is probably the best candidate for a role in the attachment of actin to membranes at the adhesion plaque. The discovery of a new protein, 215k, of unknown function, in the adhesion plaque suggests that many more proteins remain to be identified in this region. Attachment of actin filaments to other regions of the plasma membrane is also considered and a protein is described that seems to be a spectrin in brain and other tissues. The brain protein resembles erythrocyte spectrin in its physical properties, in binding actin, in being associated with cell membranes and in cross-reacting immunologically. We suggest that the brain protein and erythrocyte spectrin both belong to a family of related proteins (the spectrins) which function in the attachment of actin to membranes in many different cell types.

Actinin↗
PubMed1982

Alterations of spontaneous quantal transmitter release at the mammalian neuromuscular junction induced by divalent and trivalent ions.

The present studies have demonstrated that Ca and Sr have a similar ability to support the release of spontaneous quanta of transmitter at the rat neuromuscular junction. The foreign ions Co, Pb, Zn and La have also been shown to support spontaneous release, although the time course of the increase in miniature end-plate potentials (m.e.p.p.s) caused by these ions is much slower than that caused by Ca and Sr. However, Sr, Co, Pb and Zn were unable to sustain m.e.p.p.s for prolonged periods. High extracellular K (10--15 mM) inhibited the steady state m.e.p.p. frequency in Ca and Sr concentrations above 1 mM, and also delayed the m.e.p.p. frequency increase induced by 1 mM Pb. It is suggested that K ions directly inhibit divalent ion entry into the nerve terminal.

Animals↗