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Biomedical subjects

S Henry

Publications and source records attributed to S Henry.

At least 91 records · Page 5Linked to original sources
PubMed1992

Cytomegalovirus (CMV) antigenemia assay is more sensitive than shell vial cultures for rapid detection of CMV in polymorphonuclear blood leukocytes.

We compared the cytomegalovirus (CMV) antigenemia assay with shell vial cultures of polymorphonuclear leukocyte (PMNL)-enriched blood fractions for rapid diagnosis of CMV viremia. PMNL fractions of 280 blood specimens from 171 patients (170 solid-organ transplant recipients and 1 patient undergoing pretransplant evaluation) were inoculated in shell vial and conventional CMV cultures. A commercially available kit (CMV-vue kit; INCSTAR Corp.) was used for the CMV antigenemia assay, in which PMNL preparations were stained with monoclonal antibodies directed against the CMV protein pp65. Mixed-leukocyte blood fractions from the same blood specimens were inoculated in parallel shell vial and conventional cultures. CMV viremia (defined by the isolation of CMV in conventional cultures) was detected in 32 (13%) of 245 PMNL fractions included in the final analysis. Twenty-eight (87.5%) were also positive in the CMV antigenemia assay, whereas 22 (69%) were positive in shell vial cultures. Ten (4%) additional PMNL fractions positive only in the CMV antigenemia assay were from eight patients with active CMV infections (six patients), who had previous or subsequent episodes of CMV viremia (seven patients), or in whom CMV was isolated in cultures of simultaneously obtained mixed-leukocyte fractions (three patients). Overall, the CMV antigenemia assay was significantly more sensitive than shell vial cultures for detection of CMV in the PMNL fraction of blood leukocytes (P < 0.01, McNemar's test), and we recommend it as the method of choice for rapid diagnosis of CMV viremia.

Antigens, Viral↗
PubMed1991

Production and characterization of monoclonal antibodies that localize human thymidylate synthase in the cytoplasm of human cells and tissue.

Thymidylate synthase (TS; EC 2.1.1.45) is an important cellular enzyme that converts dUMP to dTMP, which is essential for DNA biosynthesis. In addition, TS is an important cellular target for the fluoropyrimidine cytotoxic drugs that are widely used in the treatment of solid tumors. We have generated five monoclonal antibodies against human TS using a recombinant human TS enzyme. These antibodies react specifically with human TS and display negligible cross-reactivity with other cellular proteins found in human cells. Binding affinity studies demonstrate that all antibodies form a tight interaction with recombinant human TS enzyme (Kd range = 0.3-11.0 nM). All antibodies display reactivity on enzyme-linked immunosorbent assay and immunoprecipitation. On Western blot analysis each detects a protein of approximately 36 kDa molecular mass under denaturing conditions. In addition to their reactivity on immunoprecipitation and Western analysis, two of the antibodies, TS 106 and TS 109, are reactive on immunohistochemical staining of human colon carcinoma cell lines and tissue, producing a granular cytoplasmic staining pattern. Specificity for TS is demonstrated by the lack of staining with preimmune IgG and the disappearance of the signal when the antibodies are preabsorbed with recombinant human TS enzyme. Quantitation of TS by Western blot analysis and biochemical FdUMP binding assay in 5-fluorouracil-resistant colon carcinoma cell lines (NCI H630R10, NCI H630R1) and a sensitive colon carcinoma cell line (NCI H630) revealed a 36- and 6-fold increase in TS in the resistant cell line as measured by the biochemical assay compared to a 39- and 10.6-fold increase as measured by densitometric analysis of the Western blot. These comparative studies of immunohistochemical, Western, and biochemical analyses reveal that the immunological detection of TS in human colon cell lines is a sensitive and quantitative assay. Thus the ability of these antibodies to detect TS in human cancer cells and tissue may allow measurement of TS in human tissues by quantitative immunohistochemistry in studies of drug resistance and for determination of proliferative rates.

Antibodies, Monoclonal↗
PubMed1991

Relative enrichment of the lighter 59 kDa form of glutamic acid decarboxylase in nerve endings: an immunoblotting study in pituitary neurointermediate lobe.

Previous studies established that glutamate decarboxylase (GAD) is present in the brain of higher vertebrates in two forms composed of the homodimeric association of two subunits 59 and 63 kDa, respectively, that were found to be structurally related. We have performed quantitative comparative immunoblotting of whole brain and pituitary neurointermediate lobe (NIL) extracts. While GAD in the brain is present in cell bodies and nerve endings, it is known to be contained in the NIL exclusively in nerve endings that belong to a relatively long gamma-aminobutyric acid (GABA)ergic pathway of central origin. The relative immunolabelling ratio of the 59 kDa to the 63 kDa subunit was at least 10-fold higher in the NIL when compared to whole brain extracts. Accordingly we suggest that the GAD composed of the 59 kDa subunits, which appears to be greatly enriched in GABAergic nerve endings, might represent the form of GAD on which short term activity regulation is exerted in relation to neuronal activity. It might result from the post-translational processing of the GAD formed from the 63 kDa subunits during the axonal transport.

Animals↗
PubMed1991

A technique for needle localization in paraspinal muscles with cadaveric confirmation.

Invasive electromyography (EMG) of the paraspinal muscles is useful in clinical and research settings. No technique for localization of the needle in specific fascicles has been validated. Recent descriptions of the segmented innervation of the multifidus imply that such a technique would add greatly to the EMG determination of root level of a radiculopathy. We have developed a technique for localization which relies on palpation of bony structures and needle insertion at certain angles and depths. The technique was evaluated by injecting latex dye in 199 locations in 13 cadavers. Dissection demonstrated that the technique was accurate in 91 of 112 injections into specific fascicles of the multifidus (originating from different spinous processes), 39 of 43 injections into the longissimus, and 35 of 44 injections into the iliocostalis. Certain types of errors would not have occurred with the aid of EMG in vivo. When these are added to the correct injections, accuracy improves 97%, 93%, and 82%, respectively. The technique described here should be useful for kinesiological studies, biopsies and injections, as well as for the EMG confirmation of a radiculopathy.

Back↗
PubMed1991

Multicenter trial of Collagraft as bone graft substitute.

Collagraft (Zimmer and Collagen Corporation) consists of a mixture of porous beads composed of 60% hydroxyapatite and 40% tricalcium phosphate ceramic and fibrillar collagen. When mixed with autogenous bone marrow, it serves as an effective bone graft substitute. Since September 1986, this material has been used in a multiclinic prospective trial, randomized against cancellous iliac crest autografts in the treatment of long bone fractures. To date, 267 patients have entered this study with 128 patients receiving cancellous autograft and 139 patients receiving Collagraft. At 6- and 12-month follow-ups, Collagraft appears to function as well as autogenous graft when used in the treatment of acute long bone fractures.

Adolescent↗
PubMed1990

RNA polymerase II C-terminal repeat influences response to transcriptional enhancer signals.

The large subunit of RNA polymerase II contains a highly conserved and essential heptapeptide repeat (Pro-Thr-Ser-Pro-Ser-Tyr-Ser) at its carboxy terminus. Saccharomyces cerevisiae cells are inviable if their RNA polymerase II large subunit genes encode fewer than 10 complete heptapeptide repeats; if they encode 10 to 12 complete repeats cells are temperature-sensitive and cold-sensitive, but 13 or more complete repeats will allow wild-type growth at all temperatures. Cells containing C-terminal domains (CTDs) of 10 to 12 complete repeats are also inositol auxotrophs. The phenotypes associated with these CTD mutations are not a consequence of an instability of the large subunit; rather, they seem to reflect a functional deficiency of the mutant enzyme. We show here that partial deletion mutations in RNA polymerase II CTD affect the ability of the enzyme to respond to signals from upstream activating sequences in a subset of promoters in yeast. The number of heptapeptide repeats required for maximal response to signals from these sequences differs from one upstream activating sequence to another. One of the upstream elements that is sensitive to truncations of the CTD is the 17-base-pair site bound by the GAL4 transactivating factor.

Amino Acid Sequence↗
PubMed1990

An anti-p24 monoclonal antibody shows cross-reactivity with multiple HIV-1 proteins.

We produced three murine monoclonal antibodies (mAbs) against the HIV-1 proteins. These three mAbs, namely CA-1, CA-2, CA-4, were IgG1 and all reacted with p24 on the HIV-1 Western blot. One of the mAbs, CA-4, also recognized p13, p21, p28, p29, p32, p39, p47, p55 on the Biotech/Du Pont HIV-1 Western blot strips and p21, p24, p28, p29, p39, p47, p55, p68, p80, p96; p110 on the Bio-Rad strips. CA-4 did not react with H-9 cell lysate nor with other retroviral antigens such as HTLV-1 or HIV-2 proteins. The binding of CA-4 to HIV-1 proteins was not blocked by deglycosylation. All three mAbs reacted with recombinant DNA derived capsid protein (p24) of HIV-1. These results suggest that many proteins in the HIV-1 Western blot contain antigenic epitope(s) similar to that of p24.

Animals↗
PubMed1990

Specific antiserum and monoclonal antibodies against the taurine biosynthesis enzyme cysteine sulfinate decarboxylase: identity of brain and liver enzyme.

Cysteine sulfinate decarboxylase (CSD), the putative biosynthetic enzyme for taurine, was purified 1,800-fold with a 1% yield from rat liver, where it was found to be 20-fold enriched compared with brain. The final fraction was homogeneous, as ascertained through sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse-phase HPLC. An antiserum was raised in the rabbit that (a) quantitatively immunoprecipitated CSD activity and (b) immunolabeled only one band (MW = 51,000) on an immunoblot from liver homogenate. Monoclonal antibodies were also raised that recognized the CSD protein and immunolabeled the same 51-kilodalton protein on an immunoblot from liver homogenate. In a brain extract, two CSD activities had been previously found and named CSDI and CSDII, according to their chromatographic elution patterns. We have compared the properties of CSDI from brain--the most likely enzyme involved in the biosynthesis of taurine in the brain, according to previous investigations-and CSD from liver: Both activities (a) were similarly eluted on ion-exchange and hydroxyapatite chromatographies, (b) showed the same elution pattern on gel filtration with an apparent native molecular weight of approximately 63,000, and (c) were immunoprecipitated in a strictly identical manner by the antiserum against liver CSD. Moreover, this antiserum as well as the monoclonal antibodies immunolabeled a single band (51 kilodaltons) on an immunoblot from brain CSD-enriched fraction or liver fraction. All these data show that CSDI from brain and liver CSD are the same monomeric enzyme. They also indicate that a specific antiserum against rat liver CSD has been raised that can be used for immunocytochemical visualization of CSD-containing cells in the brain.

Animals↗
PubMed1990

Non-salary retention incentives for social workers in public mental health.

While workers' reasons for leaving jobs are myriad, little is known of what might induce workers to remain in jobs. The literature reports that money, alone, is not sufficiently persuasive as an incentive. This study of social workers in the public mental health system in Colorado reveals the incentive value of a set of non-salary retention measures. The findings of the study show that measures which furthered professional enrichment, contribution to the profession, and the exercise of professional autonomy are rated most highly. Cross-tabulations with some demographic variables reveal significant findings. Recommendations for implementing the findings are presented.

Colorado↗
PubMed1990

[In vitro bactericidal activity of tobramycin and amikacin alone or in combination against Pseudomonas aeruginosa isolated from patients with cystic fibrosis].

The bactericidal kinetic of 60 P. aeruginosa isolates (40 from cystic fibrosis sputum and 20 from various origins) was studied. Liquid medium micromethod was performed. Bacteria were incubated with tobramycin and amikacin alone at several concentrations and combined with piperacillin, cefsulodin, ceftazidim, imipenem, and ciprofloxacin at concentrations obtained in vivo. When used alone, tobramycin showed the most rapid bactericidal activity, whatever the concentration used. The bactericidal activity (greater than or equal to 99.99% killing of the inoculum) was obtained in 5 hours, with 1 or 2 x MIC of the majority of the strains, with the 2 aminoglycosides. No difference was found between tobramycin and amikacin, when combinated with an antibiotic which provides a notable increase of the rapidity of the bactericidal activity. The combination of amikacin plus imipenem was more rapidly bactericidal: 48% of strains; 26% were synergistically inhibited by amikacin plus ciprofloxacin. When correlated with the susceptibility patterns of studied micro-organisms, the results were rather unpredictive.

Amikacin↗
PubMed1989

Comparison through immunoblotting of glutamic acid decarboxylase (GAD) from newborn and adult rat brain.

Immunochemical characterization of glutamic acid decarboxylase (GAD) from brain extracts of newborn and adult rats was investigated using a GAD antiserum that was previously raised against brain GAD from adult rats. According to the immunoprecipitation and saturation curves, no significant differences could be found as to the recognition of newborn and adult GAD by the antiserum. On immunoblots, both extracts revealed the same two immunolabelled bands (mol.wt. 59,000 and 62,000 +/- 2000 Da). In both cases, the lightest band showed the strongest staining. Quantitative analysis of the immunolabelling indicated that each immunolabelled band was enriched about 10-fold in the adult brain extract. These data did not reveal any difference between newborn and adult GAD that was reminiscent of the difference found in an earlier study between GAD from lower and higher vertebrates. Whatever the regulatory mechanism responsible for the presence of two forms of GAD in the adult brain, it is already fully operative in newborn animals.

Aging↗
PubMed1989

The spore coat of a fucosylation mutant in Dictyostelium discoideum.

Strain HL250 of Dictyostelium discoideum cannot convert GDP-mannose to GDP-fucose, resulting in an inability to fucosylate protein. This affects a group of proteins which are normally fucosylated intracellularly and then secreted via prespore vesicles to become part of the outer lamina of the spore coat. We have found that strain HL250 nevertheless accumulates typical amounts of these proteins, stores them normally in prespore vesicles, and secretes them normally to become a part of the spore coat. However, affected proteins are proteolyzed after germination, the spore coat is more accessible to penetration by a macromolecular probe, and germination is inefficient in older spores. These findings can be explained by a dependence of the integrity of the outer layer of the spore coat on protein-linked fucose.

Antibodies, Monoclonal↗
PubMed1989

Development of a health questionnaire specific for end-stage renal disease.

To compare the efficacy of various end-stage renal disease (ESRD) therapies valid and reproducible probes which measure well-being and are specific for ESRD are necessary. Four studies were undertaken to provide and test these probes. (1) 107 dialysis and 119 transplant recipients were interviewed to determine the prevalence of 24 physical symptoms. (2) A questionnaire was devised using 2 new indexes (a symptom scale derived from the first study using 12 symptoms and an affect scale comprising 12 emotions) and 6 indexes previously used in other chronic illnesses. Interobserver and intraobserver reproducibility was satisfactory. (3) Construct validity for the questionnaire was shown by interviewing 97 dialysis and 82 transplant patients in whom we hypothesized that physical well-being would be better in transplant patients. After age matching the transplant group was more active, with a higher objective quality of life and fewer physical symptoms than the dialysis group. (4) 63 stable dialysis, 67 stable transplant, 15 dialysis patients successfully transplanted in the intervening year and 5 failed transplanted patients were reinterviewed 1 year later to assess the responsiveness of the questionnaire. In the group who had recently been successfully transplanted both physical, affect and quality of life scores showed a major improvement following transplant. We conclude that this questionnaire is specific for ESRD, examines physical, psychological, and social well-being, is brief, easily administered, reproducible, has construct validity and is responsive to changes in therapy.

Adult↗
PubMed1989

Increased basal glucose production and utilization in children with recent obesity versus adults with long-term obesity.

To characterize the abnormalities of glucose homeostasis and insulin action early in the course of human obesity, we studied in vivo glucose kinetics in seven children who were recently massively overweight. At time of study they were gaining weight at a rate of 13.5 +/- 1.4 kg/yr. They were compared with six age-matched control subjects. Six adults with long-term obesity and five normal adults were studied in parallel. The obese children and adults were normoglycemic and hyperinsulinemic. We found that glucose production and utilization were remarkably higher in obese children (295 +/- 18 mg/min; 7.6 mg.kg-1 lean body mass.min-1) than in control children (129 +/- 13 mg/min; 4.4 mg.kg-1 lean body mass.min-1, P less than .01) and obese adults (151 +/- 8 mg/min; 3.1 +/- 0.3 mg.kg-1 lean body mass.min-1, P less than .01). Obese adults had normal rates of glucose production and utilization. Insulin- and non-insulin-mediated glucose uptake, estimated with somatostatin-induced suppression of endogenous insulin secretion, contributed almost equally to the excess glucose utilization observed in the obese children. When studied with the euglycemic-hyperinsulinemic clamp, obese children could not increase glucose disposal to the same extent as normal children and were not able to adequately suppress their endogenous glucose production. Recently obese children are therefore characterized by an increased basal glucose turnover rate and an already established insulin resistance of the liver and probably the skeletal muscles.

Adipose Tissue↗
PubMed1989

Working in private.

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Allied Health Personnel↗
PubMed1988

Clinical features and severity of nonspecific symptoms in dialysis patients.

Nonspecific symptoms are common in dialysis patients but few methods are available to measure their severity and their response to alteration in dialysis therapy. To determine the clinical features and measure the severity of the most important symptoms in end-stage renal disease (ESRD) patients, 97 dialysis patients were interviewed, 63 of whom were reinterviewed 1 year later. For comparison 82 transplant recipients were also interviewed. The six most important symptoms in dialysis patients (using the product of the patient's perception of severity and prevalence) were tiredness, cramps, pruritus, dyspnea, headaches and joint pain. The symptoms were long-standing, occurred frequently, with little difference in prevalence between hemo- and peritoneal dialysis patients, and were often unrelated to a hemodialysis session. For each symptom, several dimensions of severity were assessed including frequency, duration, effect on sleep, daily living, activity, subjective quality of life and necessity for drug therapy. Often these dimensions did not correlate with patient's perception of severity. For each symptom these items were combined to give an aggregate score with a range 0-10. Interobserver reproducibility for each symptom score was greater than or equal to 0.7 but intraobserver reproducibility was poor for 3 symptoms, because of the fluctuating nature of the symptoms. Construct validity was demonstrated by finding a significantly worse distribution of aggregate scores for tiredness, cramps, pruritus, dyspnea and nausea/vomiting in dialysis compared to transplant patients. Aggregate scores changed little after 1 year's follow-up in stable dialysis patients but significant improvement in the aggregate scores for tiredness, dyspnea and nausea/vomiting were observed in 14 patients after successful transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

Attitude to Health↗