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Biomedical subjects

D R Johnson

Publications and source records attributed to D R Johnson.

At least 163 records · Page 9Linked to original sources
PubMed1991

Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies.

Autoantibodies purified from humans catalyse the hydrolysis of the neurotransmitter, vasoactive intestinal peptide (VIP). Evidence that the hydrolysis of VIP is due to antibodies includes: the antibody preparations are free of detectable non-immunoglobulin (non-Ig) contamination; the hydrolytic activity is removed by precipitation with anti-human IgG antibody; human B lymphoblastoid cells transformed with Epstein-Barr virus secrete hydrolytic antibodies in culture; the Fab fragments of the antibodies exhibit VIP hydrolysis; and affinity chromatography on immobilized VIP permits purification of specific antibodies with greatly enriched hydrolytic and binding activities. One of the catalytic antibody preparations hydrolyses the Gln-16-Met-17 bond. Studies with synthetic VIP fragments showed that the epitope recognized by this antibody is formed by VIP(15-28). Important binding interactions are contributed by VIP(22-28), a sequence four residues distant from the scissile bond. Antibodies from a second subject hydrolyse six peptide bonds in VIP, clustered between residues 14 and 22. These bonds link amino acids of different charge, size and hydrophobicity, suggesting that the hydrolytic repertoire of the antibodies is considerable. The antibodies do not hydrolyse peptides unrelated in sequence to VIP. Cleavage of several peptide bonds in VIP by polyclonal antibody preparations may be due to several antibodies, each with a unique cleavage specificity. Alternatively, a single antibody may make catalytically productive contact at multiple peptide bonds in the substrate, because of conformational flexibility of VIP or of the antibody active site. Purified light chains from the catalytic antibodies hydrolysed VIP more rapidly than did intact antibodies. The residues constituting the catalytic site of an antibody may be encoded in germline V-region genes or may arise during maturation of the antibody response.

Amino Acid Sequence↗
PubMed1991

Medical problem solving and uncertainty in the emergency department.

STUDY OBJECTIVE: To compare the diagnostic processes of experienced emergency physicians with those of novices. DESIGN: Prospective, convenience sample of patients. SETTING: Emergency department of a county university medical center in a large southwestern urban community. PARTICIPANTS: Experienced emergency physicians (attending and senior residents) and novice clinicians (junior residents and senior medical students). INTERVENTIONS: Participants developed initial diagnostic impressions after reviewing the chief complaint, nurse triage notes, and vital signs. Tests were then selected, and a final diagnostic impression was identified after results were known. Clinicians also marked a visual analog scale corresponding to their estimate that each diagnostic possibility was correct. RESULTS: Experienced physicians increased their certainty more than novices (P = .014). They deviated from a standard history-physical-laboratory sequence more often than novices (P = .008). CONCLUSION: Expertise in medical decision making is characterized by a moderate initial level of certainty concerning a diagnosis that significantly increases as the experienced clinician follows a flexible strategy of testing to arrive at a final diagnosis.

Academic Medical Centers↗
PubMed1991

Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl-CoA:protein N-myristoyltransferase.

The S. cerevisiae myristoyl-CoA:protein N-myristoyltransferase gene (NMT1) is essential for vegetative growth. NMT1 was found to be allelic with a previously described, but unmapped and unidentified mutation that causes myristic acid (C14:0) auxotrophy. The mutant (nmt1-181) is temperature sensitive, but growth at the restrictive temperature (36 degrees C) is rescued with exogenous C14:0. Several analogues of myristate with single oxygen or sulfur for methylene group substitutions partially complement the phenotype, while others inhibit growth even at the permissive temperature (24 degrees C). Cerulenin, a fatty acid synthetase inhibitor, also prevents growth of the mutant at 24 degrees C. Complementation of growth at 36 degrees C by exogenous fatty acids is blocked by a mutation affecting the acyl:CoA synthetase gene. The nmt1-181 allele contains a single missense mutation of the 455 residue acyltransferase that results in a Gly451----Asp substitution. Analyses of several intragenic suppressors suggest that Gly451 is critically involved in NMT catalysis. In vitro kinetic studies with purified mutant enzyme revealed a 10-fold increase in the apparent Km for myristoyl-CoA at 36 degrees C, relative to wild-type, that contributes to an observed 200-fold reduction in catalytic efficiency. Together, the data indicate that nmt-181 represents a sensitive reporter of the myristoyl-CoA pools utilized by NMT.

Acyltransferases↗
PubMed1991

Stability of streptococcal pyrogenic exotoxin production with laboratory manipulation of group A streptococci.

Because of reported differences in the production of streptococcal pyrogenic exotoxins by group A strains associated with severe streptococcal infections, the stability of exotoxin production by specific strains was examined by passing group A streptococci on blood agar culture plates daily for 20 days. No changes were detected in either exotoxin genes or in exotoxin production during this time, suggesting that these reported differences are due to other explanations such as differences in the strains collected from various geographic areas or to laboratory methodologic differences.

Bacterial Proteins↗
PubMed1991

A study of facial growth in the sooty mangabey Cercocebus atys.

This study examines the pattern of facial bone growth remodelling in the sooty mangabey (Cercocebus atys) by scanning electron microscopy (SEM) of high-accuracy replicas. The efficacy of the technique is appraised by SEM interpretations of facial remodelling in the crab-eating macaque (Macaca fascicularis) and correlative histological examination. The results indicate that the distribution of depository and resorptive areas in Cercocebus closely parallels that which has been observed in Macaca. It is suggested that the different adult facial morphologies in the sooty mangabey and crab-eating macaque are the result of changes in the rates of remodelling events that may be coupled with different patterns of sutural growth (which could not be studied by SEM).

Animals↗
PubMed1991

Electronic mail and privacy: setting your company's policy.

In setting a privacy policy, a company must consider not only its interest and the interests of its employees, but also the interests of third parties, including suppliers, customers, law enforcement agencies, and others who may have a legal right to or interest in accessing company records, including electronic mail.

Civil Rights↗
PubMed1991

The effects of disorders of cartilage formation and bone resorption on bone shape: a study with chondrodystrophic and osteopetrotic mouse mutants.

The shapes of scapulae and basi-occipital bones from three genetically distinct achondroplastic mutants and one osteopetrotic mutant in the mouse (achondroplasia, brachymorphic, stumpy and grey lethal), and appropriate controls, have been compared using Fourier analysis and multivariate statistical techniques. Normal littermates were generally similar in shape, but mutants were significantly different from these controls and from each other. The pattern of morphological differences between the mutants and between the mutants and normal controls is examined. These differences are discussed in relation to the different effects of the four genes on bone morphogenesis, and the significance of these findings in relation to the contributions of cartilage formation and bone resorption to skeletal morphogenesis is considered.

Animals↗
PubMed1990

The variability of patterns of sexual dimorphism in the hominoid skull.

Univariate and multivariate statistical analyses are applied to a number of cranial dimensions and angles from living hominoids in order to investigate the patterns of sexual dimorphism in these groups. Clear differences in patterns of cranial sexual dimorphisms are demonstrated not only between genera but also within a single species (Homo). These differences overlay the common finding of a sexual size difference in all groups. The results imply that caution is required in using the sexual dimorphisms of living hominoids as models for those anticipated in fossils.

Animals↗
PubMed1990

Consecutive enzyme cascades: complement activation at the cell surface triggers increased tissue factor activity.

Complement activation at the cell surface initiates cell damage through a series of reactions occurring at the cell membrane and, after assembly of the terminal membrane attack complex, produces leakage of cytoplasmic contents from the cell. It has been documented that chemical or physical damage to cell membranes can cause a rapid increase in the expression of tissue factor procoagulant activity. In this study, antibody-mediated complement activation at the cell surface resulted in increased tissue factor activity, which correlated with cytolysis, as measured by 51-chromium release. Therefore, complement fixation on the cell surface can have a direct and immediate stimulatory effect on the coagulation cascade at the point of its initiation, with formation of a fibrin clot requiring only three consecutive proteolytic reactions after immunologically mediated cell damage.

Antibodies↗
PubMed1990

Immunovirological studies of fatal infectious mononucleosis in a patient with X-linked lymphoproliferative syndrome treated with intravenous immunoglobulin and interferon-alpha.

We have studied a 19-year-old male with X-linked lymphoproliferative syndrome (XLP) and infectious mononucleosis (IM) who was treated with high-dose immunoglobulin (500 mg/kg/day) and recombinant interferon (IFN)-alpha (2 x 10(6) IU/m2/day). Fulminant hepatitis was delayed; however, virus-associated hemophagocytic syndrome, cholestatic jaundice, and renal failure occurred terminally. Initially, nonspecific natural killer (NK) cell activity against K562 cells was normal but it gradually decreased. Although reactive T cells were markedly increased in his blood during the acute phase, spontaneous EBV-positive cell lines were easily established. Additionally, his mononuclear cells produced IFN-gamma but not IFN-alpha prior to treatment. Based on results of in vitro studies, we conclude that both IFN-alpha and IFN-gamma production are likely necessary for inhibiting EBV immortalization in vitro. Both IFN-alpha and -gamma were produced in cultures of B95-8 EBV-infected mononuclear cells from EBV-seropositive healthy individuals. These results suggest that defective EBV-specific cytotoxic T cell activity accompanied with defective or discordant IFN-alpha and -gamma production permitted the development of fatal IM in this patient. Combined treatment with immunoglobulin and IFN-alpha appeared to be partially effective during the early stage of this disease.

Adult↗
PubMed1990

Bone surface structure in osteopetrotic grey lethal (gl/gl) and microphthalmic (mi/mi) mutant mice as revealed by scanning electron microscopy.

The surface structure of bone from two genetically distinct osteopetrotic strains of mice, grey lethal (gl) and microphthalmic (mi) has been examined by scanning electron microscopy. Although both conditions produce a classical osteopetrotic phenotype the means by which this is achieved is quite different. gl mice appear to retain woven bone and show no evidence of resorption: mi mice show evidence of imperfect remodelling. These results are in accordance with what is already known of osteoclast structure and function in these mutations.

Animals↗
PubMed1990

The role of arthroscopy in the problem total knee replacement.

Fourteen patients were retrospectively reviewed to examine the role of arthroscopy in the diagnosis and treatment of the problem total knee arthroplasty (TKA), and to define parameters for indications, techniques, and results. All patients had undergone a previous TKA, and postoperatively had problems with pain and/or range of motion. Routine evaluation failed to reveal sepsis or aseptic loosening. Arthroscopy was used to evaluate and treat certain specific conditions. The arthroscope was successful in removing loose bodies, correcting patella subluxation with an arthroscopic lateral release, excising a symptomatic pseudomeniscus, and releasing intraarticular adhesions to improve motion and relieve pain. The postoperative knee score improved 73%. Arthroscopy is a valuable tool to evaluate a painful TKA, and can be used to treat certain conditions that would otherwise require an arthrotomy. The rehabilitation time is easier on the patient and much quicker. Arthroscopy of a TKA does not compromise any future procedures. However, it remains a technically demanding procedure whose indications and expectations are still being defined.

Aged↗
PubMed1990

Tumor necrosis factor and immune interferon synergistically increase transcription of HLA class I heavy- and light-chain genes in vascular endothelium.

Tumor necrosis factor and immune interferon synergistically increase cell-surface expression of class I major histocompatibility complex molecules in cultured human endothelial cells. We report that tumor necrosis factor and interferon gamma each independently increase mRNA levels and together cause a greater-than-additive (i.e., synergistic) increase in steady-state mRNA levels and transcriptional rates of the class I heavy- and light-chain genes. HLA heavy-chain mRNA is equally stable in cytokine-treated and -untreated endothelial cells. Interferon gamma does not increase tumor necrosis factor receptor number or affinity on human endothelial cells. We conclude that the synergistic increase in class I major histocompatibility complex cell-surface expression results principally from the synergistic increase in transcriptional rates. We propose that this increase is caused by the cooperative binding of independently activated transcription factors to the promoter/enhancer sequences of class I genes.

Base Sequence↗
PubMed1990

Functional analysis of protein N-myristoylation: metabolic labeling studies using three oxygen-substituted analogs of myristic acid and cultured mammalian cells provide evidence for protein-sequence-specific incorporation and analog-specific redistribution.

Covalent attachment of myristic acid (C14:0) to the NH2-terminal glycine residue of a number of cellular, viral, and oncogene-encoded proteins is essential for full expression of their biological function. Substitution of oxygen for methylene groups in this fatty acid does not produce a significant change in chain length or stereochemistry but does result in a reduction in hydrophobicity. These heteroatom-containing analogs serve as alternative substrates for mammalian myristoyl-CoA:protein N-myristoyltransferase (EC 2.3.1.97) and offer the opportunity to explore structure/function relationships of myristate in N-myristoyl proteins. We have synthesized three tritiated analogs of myristate with oxygen substituted for methylene groups at C6, C11, and C13. Metabolic labeling studies were performed with these compounds and (i) a murine myocyte cell line (BC3H1), (ii) a rat fibroblast cell that produces p60v-src (3Xsrc), or (iii) NIH 3T3 cells that have been engineered to express a fusion protein consisting of an 11-residue myristoylation signal from the Rasheed sarcoma virus (RaSV) gag protein linked to c-Ha-ras with a Cys----Ser-186 mutation. This latter mutation prevents isoprenylation and palmitoylation of ras. Two-dimensional gel electrophoresis of membrane and soluble fractions prepared from cell lysates revealed different patterns of incorporation of the analogs into cellular N-myristoyl proteins (i.e., protein-sequence-specific incorporation). In addition, proteins were identified that underwent redistribution from membrane to soluble fractions after incorporating one but not another analog (analog-specific redistribution). Comparable studies using the model RaSV-ras chimeric protein also demonstrated analog-specific differences in incorporation, varying from approximately 25% of the total RaSV-ras chimeric protein with 5-octyloxypentanoate to greater than 50% with 12-methoxydodecanoate. Modification by this latter compound was so extensive that the amount of membrane-associated N-myristoylated protein was decreased. Incorporation of each of the analogs caused a dramatic redistribution to the soluble fraction, comparable to that seen when myristoylation was completely blocked by mutating the protein's site of myristate attachment (glycine) to an alanine residue. The demonstration that these analogs differ in the extent to which they are incorporated and in their ability to cause redistribution of any single protein suggests that they may also have sufficient selectivity to be of potential therapeutic value.

Animals↗