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Biomedical subjects

B Perry

Publications and source records attributed to B Perry.

At least 37 records · Page 2Linked to original sources

Searching for the Magic Johnson effect: AIDS, adolescents, and celebrity disclosure.

The objectives of this study were to measure changes in AIDS-related attitudes and behaviors in adolescents in the 13 months following Magic Johnson's disclosure that he was HIV positive, and to test whether gender, race, age, sexual experience, and pre-existing HIV-avoidant behaviors would emerge as significant dependent variables. Adolescent clinic attendees (N = 181) ages 12-19 in four cities completed a questionnaire assessing change in AIDS-related attitudes and behaviors since Johnson's announcement. Respondents were divided into low-risk and at-risk groups. Sixty percent of respondents reported that Magic Johnson's announcement had increased their awareness of AIDS, 65.4% reported increased self-efficacy in a sexual situation, 37.2% reported that they had changed their perceived AIDS risk, 37.8% described increased resistance to peer pressure for sexual intercourse. The low-risk group was more likely to report increased self-efficacy and resistance to peer pressure but no change in perceived risk or increased AIDS awareness. Significant relationships were found between gender and increased AIDS awareness, gender and increased resistance to peer pressure to engage in sexual intercourse, race and increased AIDS awareness, and more lifetime sex partners and increased self-efficacy.

Acquired Immunodeficiency Syndrome↗

Increased activity of porcine pancreatic phospholipase A2 by designed long-range electrostatic stabilisation of the transition state.

Stabilisation of the catalytic transition state by long-range charge interactions has been tested with mutagenesis for porcine pancreatic phospholipase A2. Electrostatics calculations were used to determine locations which would interact preferentially with one part of the dipolar charge separation that is believed to develop in the transition state. Experiment shows increased enzyme activity relative to wild-type recombinant enzyme for mutants N97D and N101D, consistent with the design.

Animals↗

Regional renal blood flow measurements using radioactive microspheres in a chronic porcine model with unilateral vesicoureteral reflux.

95Niobium labeled radioactive microspheres were used to determine regional renal blood flows in a porcine model of chronic sterile vesicoureteral reflux. Unilateral vesicoureteral reflux was surgically created in 5 mini-pigs and regional renal blood flows were determined by microsphere injection 6 months later. The contralateral nonrefluxing kidney acted as a control. There was a significant reduction of flow in the inner cortical regions of the middle (78% of control, p < or = 0.0437) and lower poles (69% of control, p < or = 0.0274), and the juxta-medullary cortical region of the lower pole (67% of control, p < or = 0.0124). There was no difference in flow in the other regions or when comparing whole kidneys. There were no differences between refluxing and nonrefluxing kidneys when comparing ratios of inner to outer cortical flow level by level. These observations are in contrast to those in acutely created vesicoureteral reflux in a porcine model, which had no significant differences in flow in any region using the microsphere technique. Decreases of blood flow in certain cortical regions may help explain some of the physiological changes in vesicoureteral reflux in children and experimental models of reflux.

Animals↗

Making a small enzyme smaller; removing the conserved loop structure of hen lysozyme.

Engineering a smaller lysozyme is a challenge for both random and site-directed mutagenesis. This paper illustrates the power of knowledge-based protein engineering in the design of a smaller lysozyme that folds correctly and has activity against bacterial cell walls. In this smaller lysozyme the conserved disulphide bridged loop is replaced by a short loop. The long loop was selected because it buries a predominantly hydrophilic surface. The short loop was discovered by searching for appropriate fragments in the protein databank. This approach is important in the design of small enzymes useful to the food industry.

Animals↗

Physiological consequence of expression of soluble and active hen egg white lysozyme in Escherichia coli.

Hen egg white lysozyme was expressed as a protein fusion with the OmpA signal sequence and an octapeptide linker in Escherichia coli. The expression yielded soluble and enzymatically active lysozyme. Lysozyme activity was detected in the periplasmic space, in the cytosol and in the insoluble cytosolic fraction of E. coli. The results indicate that the environmental conditions in both the cytosol and the periplasmic space of E. coli were sufficient for correct protein folding and disulphide bond formation of eukaryotic recombinant lysozyme. However, the expression of active enzyme in E. coli consequently led to bacterial cell lysis due to hydrolysis of the peptidoglucan.

Animals↗

Modification of the stability of phospholipase A2 by charge engineering.

Electrostatic interactions play an important role in stabilizing the folded conformation of globular proteins. Here we predict the change in stability of charge engineered mutants, construct these mutants and compare the predicted change in stability with that observed. The change in stability was correctly predicted for two of the three mutants and the factors responsible for the discrepancy between observation and prediction for the third mutant are discussed.

Enzyme Stability↗

Production and characterisation of monoclonal antibodies to parathyroid hormone (1-34).

Monoclonal antibodies to the biologically active N terminal region of parathyroid hormone (PTH) suitable for use in the measurement of circulating PTH concentrations have proved difficult to produce. In this study, no serum PTH antibody titres could be detected in mice using synthetic human PTH (1-34) (free or coupled to albumin) or PTH (1-10) (coupled to keyhole limpet haemocyanin) as immunogen. A consistent response to PTH (1-34) peptide was obtained in DA rats. We have produced five monoclonal antibodies to PTH (1-34) derived from the fusion of DA rat spleen cells and the mouse myeloma line X63 Ag.8.653. Bulk production of the antibodies was achieved using congenitally athymic mice for ascites production. Antibody assessment studies revealed the antibodies to be sensitive to the oxidation state of the methionine residues in PTH (1-34). Two of the antibodies, 3B3 and 6E3, were shown to be of potential use in measuring circulating PTH (1-84) when used in combination with available antibodies to C terminal PTH. A third antibody, 4G3, which failed to recognise PTH (1-84) when used in combination with 3B3, formed the basis of a specific assay for PTH (1-34).

Animals↗

Renal blood flow measurements using radioactive microspheres in a porcine model with unilateral vesicoureteral reflux.

Renal blood flow was studied in Yorkshire pigs with surgically created unilateral vesicoureteral reflux using radionuclide labeled microspheres. The contralateral, nonrefluxing kidney served as a within-subject control and a group of nonoperated pigs without reflux served as an additional control group. Unilateral reflux did not change the distribution of whole renal blood flow between the refluxing and nonrefluxing kidneys. In addition, reflux had only minor effects upon the distribution of blood flow to cortical regions within the kidneys.

Animals↗

Specificity of two-site immunometric assays.

The assessment of specificity in two-site immunometric assays is complex. Conventional approaches based on standard substitution with cross-reactant may yield misleading results. An 'in-house' immunoradiometric assay (IRMA) for human growth hormone (hGH) appeared free from cross-reactivity with human placental lactogen (hPL) in a single point standard substitution experiment performed by the United Kingdom External Quality Assessment Scheme (UK EQAS). However, detailed studies based on the substitution of hGH standards with hPL cross-reactant, the addition of hPL cross-reactant to hGH standards and the recovery of added hGH from endogenous cross-reactant (sera from pregnant subjects) revealed gross cross-reactivity of hPL in the hGH assay. We recommend that the validation of assay specificity should require the demonstration of quantitative recovery of a range of analyte concentrations from human sera that contain varying endogenous concentrations of cross-reactant.

Calibration↗

A two-site immunoradiometric assay for PTH(1-84) using N and C terminal specific monoclonal antibodies.

We report the development of a two-site immunoradiometric assay for PTH(1-84) based on two site-specific monoclonal antibodies--3B3 (radiolabelled antibody) specific for PTH(1-34) and ESQ1 (on solid phase) specific for PTH(74-84). Antibody 3B3 is sensitive to the oxidation of the methionine residues in PTH(1-34) therefore hydrogen peroxide (0.1 M) is added to the incubation mixture. Validation studies confirm quantitative recovery of both oxidized and reduced PTH(1-84). The assay has a minimum detection limit of 0.5 pmol/L and a range of 1.5-250 pmol/L with an intra-assay CV of less than 10% (2.8-250 pmol/L less than 5% CV). Studies on clinical samples indicate good discrimination between normal subjects (mean 2.21; range 1.0-5.0 pmol/L) and patients with primary hyperparathyroidism (mean 21.0; range 5.8-100 pmol/L) who in turn are well separated from patients with hypercalcaemia of malignancy (14/18 less than 0.5 pmol/L).

Adult↗

Ovulation, fertilization and early embryonic development in the bitch (Canis familiaris).

Using circulating plasma hormone estimations, ovulation was monitored in bitches. The results obtained indicate that the timing of ovulation bears little relationship to alterations in sexual behaviour. The bitches were killed and reproductive tracts were removed at various intervals after ovulation and ova or embryos were recovered. The embryo stages were assessed visually and some were investigated histologically. Embryonic development, to early blastocyst stage, took place within the oviducts during the first 12 days after ovulation and there was a marked increase in size between the early and late blastocyst. A culture system using cells from the uterine tube supported the development of one 1-cell embryo to the morula stage.

Animals↗

Pharmacokinetics and effects of recombinant human erythropoietin after intravenous and subcutaneous injections in healthy volunteers.

A double-blind, placebo-controlled study of the pharmacokinetics and safety of multiple doses of recombinant human erythropoietin [rHuEPO 150 or 300 U/kg either by intravenous (IV) bolus or subcutaneously (SC)] in normal male subjects demonstrated that rHuEPO had a dose-related effect on the hematocrit independent of the route of administration and that multiple doses of rHuEPO had no direct pressor effects. When rHuEPO was injected IV, a monoexponential decrease in serum EPO level was evident for 18 to 24 hours postdose. Absorption of SC injected rHuEPO occurred more slowly, with relatively low serum EPO levels being maintained for 48 hours. All rHuEPO antibody titer determinations were negative. With the exception of significant increases in hemoglobin and hematocrit, no clinically significant changes occurred. No hypertensive, convulsive, or thrombotic events were observed. Of the adverse experiences observed in 10 subjects, none was considered clinically significant, and none of the subjects dropped out because of adverse experiences.

Adult↗