Background Gonorrhoea notifications have increased substantially in Australia over the past decade. Neisseria gonorrhoeae is already highly resistant to several antibiotics and so, alternatives to first-line treatment are generally strongly discouraged. The penicillin allergy label (AL) on patient medical records has previously been shown to influence prescribing practices, to the detriment of best-practice management and antimicrobial stewardship.
View Article and Find Full Text PDFThis review article discusses dental practice implications of prion diseases, including Creutzfeldt-Jakob disease. The current universal precautions used for infection control in dentistry do not inactivate infectious prions. There is a theoretical, yet real risk of prion disease transmission through dental treatment, although the magnitude of that risk has not yet been determined.
View Article and Find Full Text PDFHuman proliferation-associated protein p120 has previously been shown to localize to the nucleolus, and several functional domains of p120 have been elucidated. By using a nitrocellulose filter binding assay and a Northwestern blotting procedure this study shows that recombinant p120 binds to an rRNA fragment in vitro with a dissociation constant of 4 nM. The specific RNA-binding region of p120 (residues 1-57) was identified with glutathione S-transferase-fused p120 deletion constructs and Northwestern blotting procedures.
View Article and Find Full Text PDFObjective: To investigate a high prevalence of systemic sclerosis (SSc; scleroderma) in a well-defined population of 21,255 Choctaw Indians residing in 8 southeastern Oklahoma counties who were "users" of Indian Health Services.
Methods: A case-control study of 12 SSc cases and 48 matched non-SSc controls (4 per case) was conducted to investigate potential occupational, residential, and infectious exposures, as well as genetic factors which might predispose to SSc. HLA class II alleles were determined by DNA oligotyping, and class I and III alleles were defined serologically.
Nucleolar p120 is a proliferation-associated protein, which becomes detectable early in the G1 phase of the cell cycle and peaks early in the S phase. A variety of human malignant tumor cells contain much higher levels of p120 than normal resting cells. The cellular functions of p120 are unknown, and little information is available on the structural characteristics of the human p120 protein.
View Article and Find Full Text PDFThe distribution of immunoreactive transforming growth factor-alpha (TGF-alpha) was studied in non-neoplastic human major and minor salivary glands using an immunoperoxidase assay in conjunction with an antiserum to human TGF-alpha. The ductal cell components of all major and minor salivary glands were found to contain significant amounts of TGF-alpha immunoreactivity. In contrast, acinar and myoepithelial cells consistently lacked immune reaction product in both types of glands.
View Article and Find Full Text PDFDiversity of cell lineages within glandular organs is generated postnatally by differentiation of committed progenitor cells. Fundamental regulatory aspects of this process are not understood. The mouse submandibular salivary gland (SSG) served as model to assess the role of epidermal growth factor (EGF) receptor signaling during emergence of cell lineage diversity.
View Article and Find Full Text PDFNucleolar phosphoprotein p120 is a low abundance, proliferation-associated protein. Several functional domains have been characterized and are discussed here such as the antigenic domain recognized by a monoclonal antibody, the nuclear/nucleolar localization domain, phosphorylation domains of casein kinase II (CKII) and protein kinase C, a putative methylation domain and an RNA binding region. By sucrose gradient sedimentation analyses, protein p120 was shown to rapidly sediment with 60-80 S pre-rRNP particles but sedimented more slowly when treated with RNAse or salt suggesting binding to RNA.
View Article and Find Full Text PDFBiol Chem Hoppe Seyler
April 1994
Topoisomerase I purified from HeLa cells was phosphorylated in vitro with protein kinase NII (pkNII) purified from calf thymus: this phosphorylation was inhibited by heparin. A peptide containing a sequence corresponding to a putative pkNII phosphorylation site in topoisomerase I was synthesized and phosphorylated with pkNII. HPLC and two-dimensional analysis show identity between the synthetic phosphorylated peptide and one topoisomerase I phosphopeptide indicating Ser10 as one of the in vitro pkNII phosphorylation sites in topoisomerase I.
View Article and Find Full Text PDFRecognition of distinct cell phenotypes within a given organ is important in defining cell relationships during development and in analyzing the role of cell-cell and cell-matrix interactions in growth and differentiation. Phenotypic definition of dissociated heterogeneous cell populations is also essential for studies on mechanisms regulating expression of cell lineage-specific gene products. Mouse submandibular salivary gland (SSG) cell phenotypes in the course of differentiative transitions in vivo and after enzymatic dissociation in primary culture were defined with monoclonal antibodies (MAb) to mammary epithelial cells and polyclonal antibodies to functional cell products.
View Article and Find Full Text PDFTopoisomerase I was phosphorylated in vitro by protein kinase C (PKC) purified from rat brain with high affinity (Km about 0.1 microM). Tryptic phosphopeptide mapping indicated that two major topoisomerase I peptides phosphorylated in vivo were comigrating with minor peptides phosphorylated by PKC in vitro.
View Article and Find Full Text PDFPrevious studies in Caucasians with progressive systemic sclerosis (PSS) have suggested associations of antitopoisomerase I (antitopo I) autoantibodies with either serologically defined HLA-DR2 or DR5. To better define class II HLA associations with the antitopo I response, 161 PSS patients (132 Caucasians and 29 American blacks) were studied for antitopo I autoantibodies by immunodiffusion and immunoblotting, and their HLA-DRB1, DRB3, DQA1, and DQB1 alleles were determined by restriction fragment length polymorphic analysis and DNA oligotyping. Among Caucasians with antitopo I, HLA-DR5(DRB1*1101-*1104), DRB3*0202 and DQw3 (DQw7,8,9) were significantly increased in frequency.
View Article and Find Full Text PDFArthritis Rheum
February 1992
Objective: To determine demographic differences in scleroderma-related autoantibodies.
Methods: One hundred fifty-six patients with systemic sclerosis were prospectively examined for anticentromere antibodies (ACA), anti-topoisomerase I (anti-topo I, or Scl-70), antinucleolar, and anti-U1 RNP autoantibodies.
Results: ACA was found in 36% of Caucasians and 4% of American blacks (P = 0.
The pentapeptide pyroGlu-Ala-Glu-Ser-Asn has been synthetized and phosphorylated in vitro at level of serine by protein kinase NII isolated from calf thymus chromatin. It is noteworthy that the calf thymus kinase NII shows a remarkable affinity for this peptide. The [32P]peptide is able to bind to several DNAs in the presence of Mg2+ (lambda phage, calf thymus, pBR540 plasmid).
View Article and Find Full Text PDFTopoisomerase cDNA and various fragments thereof generated by the DNA polymerase chain reaction were cloned into plasmid expression vectors (pET series) and the expressed polypeptides were probed with scleroderma sera from seven different patients immunoreactive with topoisomerase I. All sera reacted selectively with a region between amino acid residues 405 and 484 of human topoisomerase I. This conclusion is based on loss of reactivity when this region was omitted from larger pieces.
View Article and Find Full Text PDFNucleophosmin (B23) was phosphorylated in vitro with [gamma-32P]ATP and a nuclear kinase (type II) purified from HeLa cells. The phosphorylation was inhibited by heparin and by 2,3-diphosphoglycerate. Peptide mapping analysis indicated that the phosphorylation site in vitro was identical to that in vivo.
View Article and Find Full Text PDFThe interactions between differentiation-associated cellular events in the intact mammary gland or in cultured mammary cells and the post-transcriptional activity of the endogenous mouse mammary tumor virus (MMTV) loci were investigated. The transcriptional activities of the endogenous MMTV proviruses of the BALB/c mouse strain (Mtv-6, Mtv-8 and Mtv-9) appear to be regulated differentially during pregnancy-induced mammary gland development (J.E.
View Article and Find Full Text PDFIn Vitro Cell Dev Biol
January 1990
The adult mouse submandibular salivary gland provides a good model system to study gene regulation during normal and abnormal cell behavior because it synthesizes functionally distinct products ranging from growth factors and digestive enzymes to factors of relevance to homeostatic mechanisms. The present study describes the long-term growth and differentiation of submandibular salivary epithelial cells from adult male mice as a function of the culture substratum. Using a two-step partial dissociation procedure, it was possible to enrich for ductal cells of the granular convoluted tubules, the site of epidermal growth factor synthesis.
View Article and Find Full Text PDFIn the course of screening cDNA expression libraries with a monospecific polyclonal antibody to topoisomerase I, we isolated three different immunopositive cDNA clones. By comparing their derived amino acid sequences, a consensus region of similarity in otherwise completely dissimilar sequences was identified as an epitope. The approach described here should identify both continuous and discontinuous topological epitopes.
View Article and Find Full Text PDFClonal populations were isolated from the mouse mammary cell line, COMMA-D, by transfection with a dominant-selectable gene, pSV2Neo, which confers resistance to the antibiotic, G418. Seven of twenty-four clones isolated retained the ability of the parental line to repopulate cleared mammary fat pads in vivo as ductal-alveolar hyperplasias. Two sublines designated CDNR2 and CDNR4 retained hyperplastic growth potential after multiple passages in vitro with low incidence of tumor formation.
View Article and Find Full Text PDFMinimum substrate requirements for nuclear NII kinase (casein II kinase) were analyzed with synthetic peptides modeled according to amino acid composition of phosphopeptides isolated from chromatin. Uncharged blocked peptide termini decreased the requirement for acidic clusters neighboring the phosphate acceptor amino acid (serine) such that only one group immediately N-terminal to serine was sufficient for kinase activity. Studies on peptide interaction with DNA showed that the model phosphopeptides bound to DNA only in the phosphorylated form suggesting involvement of phosphorylation in protein-DNA interactions yet to be identified.
View Article and Find Full Text PDFIn the course of development of a "library" of monoclonal antibodies to nucleolar proteins, a monoclonal antibody to a nuclear antigen with a molecular weight of 150,000 was obtained. Using this monoclonal antibody as an immunocytochemical probe, low immunofluorescence was demonstrated in human peripheral blood lymphocytes or HL-60 cells treated with retinoic acid. In contrast, a high degree of immunofluorescence was detected in rapidly proliferating human cells (HeLa, Hep-2, HL-60), in Novikoff hepatoma cells, and in phytohemagglutinin-activated human blood lymphocytes.
View Article and Find Full Text PDFBiochem Biophys Res Commun
July 1988
The partial amino acid sequence of rat topoisomerase I was determined by gas-phase microsequencing. Seven tryptic peptides closely matched the sequences deduced from human topoisomerase I cDNA (94.5% homology).
View Article and Find Full Text PDFPurified anti-topoisomerase I immunoglobulin G (IgG) was microinjected into nuclei of Chironomus tentans salivary gland cells, and the effect on DNA transcription was investigated. Synthesis of nucleolar preribosomal 38S RNA by RNA polymerase I and of chromosomal Balbiani ring RNA by RNA polymerase II was inhibited by about 80%. The inhibitory action of anti-topoisomerase I IgG could be reversed by the addition of exogenous topoisomerase I.
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