Biomedical subjects

J R Turner

Publications and source records attributed to J R Turner.

At least 19 recordsLinked to original sources

PubMed1992

Substrate specificity of isopenicillin N synthase.

Highly purified isopenicillin N synthase (IPNS) from two sources (naturally occurring in Penicillium chrysogenum and that expressed in Escherichia coli via a cloned gene derived from Cephalosporium acremonium) have been isolated and utilized in vitro to test synthetic modifications of the natural substrate, (L-alpha-amino-delta-adipyl)-L-cysteinyl-D-valine (ACV). A very sensitive procedure utilizing the ability of beta-lactams to induce the synthesis of beta-lactamase was employed to determine whether an ACV analogue could serve as a substrate for IPNS. A wide variety of amino and carboxyl terminal tripeptide substitutions were examined and found to elicit positive beta-lactamase induction profiles. However, none of these modifications were found to function as efficiently as a substrate as ACV. One of the beta-lactam products which was formed from the reaction of IPNS and the tripeptide analogue was independently synthesized and evaluated for antibacterial activity. Modification of the L-cysteine residue in the second position of ACV resulted in tripeptides that were unable to serve as substrates. Conversion of the D-valine residue in the third position of ACV to an aromatic amino acid or to a highly electronegative residue such as trifluorovaline resulted in elimination of substrate activity and creation of an inhibitor of the enzyme.

Amino Acid Sequence↗

PubMed1992

Stress-induced changes in the rate of sodium excretion in healthy black and white men.

The effects of exposure to competitive mental stressors on rates of excretion of sodium (Na), potassium (K) and fluid, together with cardiovascular responses, were evaluated in 14 black and 14 white normotensive men after 3 days of controlled diet (200 mEq Na, 100 mEq K daily). In the first hour of a 5-hr protocol, subjects ingested a standard dinner and drank 1 liter of water. In hours 2-5, they voided to provide urine collections every 30 min, and drank 200 ml additional water after each collection. Hours 1-2 were for stabilization of excretion, hour 3 was baseline, hour 4 was stress and hour 5 was post-stress rest. During stress, 20 men showed faster natriuresis and 8 showed slower natriuresis than at baseline. These subgroups did not differ in any excretion measure at baseline. During stress, the slow stress natriuresis group showed slower excretion of potassium and fluid as well as sodium; slow natriuresis and kaliuresis persisted 30 min after stress. Creatinine clearance rate was also decreased, but only during the first 30 min of stress. Slow stress natriuresis subjects were found to have higher blood pressures during baseline and stress, greater heart rate and cardiac output increases during stress, and a larger percentage had hypertensive parents (63 vs 37%) compared to fast stress natriuresis subjects. Slow stress natriuresis was observed in 50 vs 17% of men with resting diastolic levels above vs below 80 mmHg, and in 43% of blacks vs 18% of whites tested. Overall, black subjects tended to excrete sodium more slowly than whites at baseline, and showed significantly lesser increases in sodium excretion rate when pressures rose during stress.

Adult↗

PubMed1992

Reactivity to stress and psychological adjustment in adults with pituitary insufficiency.

OBJECTIVE: Hypopituitary adults who were affected during childhood have a below-average rate of marriage, a rate of unemployment that exceeds national norms, and often indicate dissatisfaction with their life circumstances. We undertook the present study to determine the effects of short stature versus those of pituitary hormone deficiency. DESIGN AND PATIENTS: We compared hypopituitary adults (n = 25) with normal short adults (n = 25) who were matched for height, sex, age and socioeconomic status. MEASUREMENTS: In these two groups of subjects, we compared the physiological responses to a simulated social stressor, a public speaking task, and we measured the psychometric attributes that are indicators of social assertiveness and extraversion. RESULTS: Before, during and after the stress of public speaking, patients with multiple pituitary hormone deficiencies (n = 20) had lower mean systolic and diastolic blood pressures than controls, while patients with isolated growth hormone deficiency (n = 5) were equivalent to controls. The reactivity to stress, assessed using delta scores based on changes in blood pressure and heart rate, was also decreased in multiple hormone deficient patients. Psychometric test results indicated that patients with multiple hormone deficiencies showed lower openness, lower assertiveness, greater neuroticism and a tendency towards less extraversion than did controls. The responses of patients with isolated GH deficiency on the psychometric tests were not statistically different from controls, but the number of subjects in this group (n = 5) was too small to draw conclusions. CONCLUSIONS: The impaired cardiovascular responses to stress in patients with multiple hormone deficiencies, compared to short control subjects and to patients with GH deficiency leads us to conclude that factors other than stature and GH are responsible for these observations. The differences might be related to insufficiency of catecholamines or cortisol in the patients with multiple hormone deficiencies. These hormonal deficiencies might also account for the socially inhibited behaviour of these patients. Our results suggest that more attention needs to be directed at preparing hypopituitary patients for the challenges of adulthood. Also, these patients may be helped by more intensive efforts at hormonal replacement in adulthood.

Adolescent↗

PubMed1992

Characterization of a novel regulatory gene governing the expression of a polyketide synthase gene in Streptomyces ambofaciens.

A key step in the biosynthesis of macrolide antibiotics is the assembly of a large macrocyclic lactone ring by a multienzyme protein complex called the polyketide synthase. In the species Streptomyces ambofaciens, the polyketide synthase for the assembly of the 16-membered ring of the macrolide antibiotic spiramycin is encoded by the biosynthetic gene srmG. Here we show that the accumulation of transcripts from the srmG promoter is governed by the regulatory gene srmR, whose predicted product, a 65 kDa polypeptide, is not significantly similar in its deduced amino acid sequence to that of previously reported proteins in the protein databases. The srmR gene product is also required for the accumulation of transcripts from srmX, an additional gene in the vicinity of srmR, but not for the accumulation of transcripts from srmR itself. Interestingly, mutations in srmR prevent the accumulation of transcripts from the spiramycin resistance gene srmB, but this is an indirect consequence of the failure of srmR mutants to produce spiramycin, which is an inducer of its own resistance gene. The possibility that srmR is the prototype for a new class of regulatory genes governing early events in the biosynthesis of macrolide antibiotics is discussed.

Amino Acid Sequence↗

PubMed1992

Comparison of impedance cardiographic measurements using band and spot electrodes.

The comprehensive assessment of cardiac function using impedance cardiography has led to increasingly widespread use of the technique in psychophysiology. Disposable adhesive band electrodes have been the most widely used electrode type, but spot electrode configurations present attractive alternatives in terms of convenience and subject comfort. The present study was designed to evaluate whether one such spot electrode configuration yielded the same information as the more standard band electrodes for cardiac output and systolic time interval measurement. Male and female healthy adult subjects (N = 20) were tested. Comparisons between spot and band electrodes were made for the absolute magnitude of cardiac output and systolic time intervals, as well as for responses to the highly reproducible effects of bicycle exercise. Consistent with previous findings, systolic time interval measurements were unaffected by electrode type. However, for cardiac output measurements, differences between spot and band electrode measurements were found. Under resting conditions, the absolute magnitudes of cardiac output values measured using spot electrodes were smaller than for band electrodes. Subtle, yet significant differences were also found for cardiac output responses to exercise, with spot electrodes indicating greater increases in cardiac output than band electrodes. At the same time, anticipated gender differences found for cardiac output at rest and in response to exercise were unaffected by electrode type. Overall, these findings suggest that when comparing the results of studies that have utilized different impedance electrode types, it would be prudent to remain alert to the possibility of confounding influences.

Adult↗

PubMed1992

Job strain and ambulatory work blood pressure in healthy young men and women.

The effect of high job strain (defined as high psychological demands plus low decision latitude at work) on blood pressure was determined in 129 healthy, nonhypertensive men (n = 65) and women (n = 64). Blood pressure measures included mean screening levels obtained in a clinical environment, mean ambulatory levels from one 8-hour workday, and the change in levels from screening to mean work levels. In male workers, men with high and low job strain showed similar blood pressures at screening, but men with high job strain showed greater increases from screening to work, resulting in higher mean work blood pressure. Occupational status was unrelated to job strain or blood pressure in men. In female workers, women with high and low job strain did not differ in any measure of blood pressure; however, there were trends for higher occupational status and greater skill discretion to be associated with higher blood pressure responses at work in women.

Adolescent↗

PubMed1991

Intracellular sites for storage and recycling of C3b receptors in human neutrophils.

Upon activation of human neutrophils, C3b receptors (type 1 complement receptors, CR1) are rapidly translocated from an intracellular pool to the surface, increasing plasma membrane expression 6- to 10-fold. This is followed by reinternalization and degradation of the receptors, even in the absence of ligand. Upregulation of surface CR1 may occur without exocytosis of primary or secondary granules, and intracellular CR1 in resting neutrophils does not sediment with these granules on density gradient fractionation. To directly localize the intracellular pools of CR1, we used immunoelectron microscopy of fixed, permeabilized cells. In resting neutrophils, CR1 is associated with the membranes of smooth-surfaced, empty-appearing vesicles whose irregular borders clearly distinguish them from primary and secondary granules. After activation by fMet-Leu-Phe, the intracellular pool is primarily found in large, conspicuous, multivesicular bodies. These bodies also incorporate colloidal gold from the extracellular fluid, suggesting that they are formed by endocytosis. Thus, the sites in which CR1 is stored in resting cells, and recycled in activated cells, are structurally unique and morphologically distinguishable.

Cell Compartmentation↗

PubMed1991

Genetics of Type A behavior in two European countries: evidence for sibling interaction.

Young male twins in The Netherlands and England completed the Jenkins Activity Survey (Dutch and English versions, respectively), a measure of Type A behavior. Separate model fitting analysis revealed a similar pattern of variance estimates and associated goodness of fit across the two countries. The data were then analyzed concurrently, with a scalar parameter included to account for differences in variance due to the disparity of the measurement scales. A model including additive genetic and individual environmental effects gave a good explanation to the data. The heritability estimate was 0.28. Models of social interaction and dominance explained the data even better, the former being preferred. The twins' parents were included in the analysis to examine population variation for Type A behavior intergenerationally. There was evidence for individual environmental experiences having a greater influence on Type A behavior in the older generation.

Adolescent↗

PubMed1991

Postural effects on blood pressure reactivity: implications for studies of laboratory-field generalization.

This study was designed as an in-laboratory evaluation of the significance of postural blood pressure (BP) adjustments in the investigation of laboratory-field generalization. Blood pressure responses were monitored while 20 subjects completed a mental arithmetic task in two postures, seated and standing. Baseline data were also collected in each posture. The standing challenge was regarded as a laboratory representation of real-world standing challenges. Reactivity scores for the seated stressor were calculated as seated task level minus seated baseline level. For the standing stressor, they were calculated in two ways; standing task level minus standing baseline, and standing task level minus seated baseline. It was hypothesized that the standing reactivity scores calculated from standing baselines (i.e. from the same-posture baselines) would be more highly correlated with seated reactivity than would the standing reactivity scores calculated using sitting baselines. This was not found to be the case for either SBP or DBP. Absolute task SBP levels were very similar in both postures, while they were significantly different for DBP, with standing task levels being higher. It was concluded that ignoring posture will not mask SBP reactivity associations between laboratory and field, but that it may do so for DBP, particularly in future studies as our overall understanding of laboratory-field generalization increases.

Adolescent↗

PubMed1991

Generalization of cardiovascular response: supportive evidence for the reactivity hypothesis.

Cardiovascular responses were monitored while 36 subjects completed a battery of laboratory stressors comprising mental arithmetic, a reaction time task, a speech task, and the forehead cold pressor. Inter-task consistency was assessed for each of 6 physiological parameters for all task pairings. Considerable inter-task consistency for reactivity scores was seen among the psychological stressors for all variables. The question of such consistency between the cold pressor and the psychological tasks was then addressed. The pattern of consistency was not as clear-cut in this case. For systolic blood pressure and pre-ejection period, reactivity scores to the cold pressor did not correlate with those to any of the psychological tasks. In contrast, cardiac output and total peripheral resistance responses showed considerable consistency. The importance of determining the nature of the relationship between psychological and physical stressors is discussed.

Adult↗

PubMed1991

A twin study approach towards understanding genetic contributions to body size and metabolic rate.

The genetic and environmental determinants of a brief assessment of metabolic rate at rest and under psychological stress were studied in 40 pairs of monozygotic and 40 pairs of dizygotic young adult male twins. Height, weight and age were employed as covariates. Univariate analyses showed a high heritability for height and weight and moderate heritability for metabolic rate. Classical twin analyses and multivariate genetic modeling indicated that genetic influences on resting metabolic rate were entirely explained by body weight: there was no independent genetic contribution to resting metabolic rate. Metabolic rate under psychological stress, on the other hand, showed a significant genetic effect. The exponent (3/4) in the power function relating body weight to resting metabolic rate was the same as that found in a wide variety of animal species, a value that has been proposed as defining a body weight set point. We speculate that an adult body weight set point is genetically transmitted. Independent genetic effects on resting metabolic rate would be observed only when the normal equilibrium between body weight and metabolic rate is unbalanced during development, aging or disease. The study illustrates the use of multivariate genetic analyses of twin data which may be readily applied to widely used metabolic rate assessments.

Adolescent↗

PubMed1990

Cardiovascular responses to behavioral stressors: laboratory-field generalization and inter-task consistency.

Blood pressure and heart rate responses were monitored while 30 medical, dental or graduate students completed four laboratory tasks. On a second day, subjects wore an ambulatory blood pressure monitor, and during this day they completed the real-world challenge of presenting their research in front of a small audience. Heart rate and blood pressures correlated significantly between all four laboratory tasks for both absolute levels and reactivity scores (calculated as task level minus pre-task baseline level). However, while heart rate and blood pressure absolute levels for each task correlated significantly with those attained during the real-world stressor, correlation coefficients obtained when similarly comparing reactivity scores were uniformly non-significant. Laboratory-field generalization was thus evident only for absolute values. Consideration was then given to the fact that the laboratory tasks were undertaken in a seated position, while subjects stood during the real-world task. Activity diaries completed by subjects during the time they wore the ambulatory monitor were examined to search for readings that could be regarded as standing baselines. Such values were obtained for 12 of the subjects. Real-world reactivity scores were recalculated using these standing baselines, and compared again with reactivity scores during the laboratory tasks. Marked increases in correlation coefficients were obtained for systolic and diastolic pressure, but not for heart rate.

Adult↗

PubMed1990

Temporal stability of the hemodynamics of cardiovascular reactivity.

Cardiovascular responses to a competitive reaction-time task were monitored in 13 male subjects tested twice, 3 months apart. The temporal stability of blood pressure responses was in line with previous reports. However, in this study impedance cardiography permitted the investigation of the hemodynamic adjustments underlying the observed blood pressure responses. Analyses revealed that cardiac output and total peripheral resistance responses displayed temporal stability, indicating that subjects' blood pressure responses on the two occasions were the result of similar hemodynamic responses. These data thus extend the literature by demonstrating that the hemodynamic response pattern itself represents a stable individual difference variable.

Adult↗

PubMed1990

Cytologic assessment of nuclear and cytoplasmic O-linked N-acetylglucosamine distribution by using anti-streptococcal monoclonal antibodies.

Recent studies have demonstrated the existence of single O-linked N-acetylglucosamine (O-GlcNAc) residues on cytoplasmic and nuclear glycoproteins. Labeled lectin and enzymatic techniques have been used to identify O-GlcNAc-bearing proteins, but no antibodies generally reactive with such O-linked GlcNAc moieties have been described. We have previously characterized monoclonal antibodies (mAbs) specific for the GlcNAc residues of streptococcal group A carbohydrate, which is composed of a polyrhamnose backbone with GlcNAc side chains. We now report that these mAbs recognize O-GlcNAc-bearing proteins. By immunofluorescence, the mAbs reacted strongly with the nuclear periphery and nucleoplasm of mammalian cells and stained the cytoplasm less intensely. The distribution was not consistent with labeling of the endoplasmic reticulum, Golgi complex, or plasma membrane. Furthermore, the staining pattern of a mutant cell line, which retains terminal GlcNAc residues on many N-linked glycans, was indistinguishable from that of wild-type cells. Nuclear and cytoplasmic staining were inhibited by free GlcNAc and were completely abolished by galactosylation of terminal GlcNAc residues. Indirect ELISA demonstrated GlcNAc- and galactosylation-inhibitable binding of the mAbs to a 65-kDa human erythrocyte cytosolic protein known to contain O-GlcNAc. Thus, these mAbs react with O-GlcNAc without apparent influence of peptide determinants, do not show detectable binding to N- or O-glycans, and, therefore, represent a valuable tool for the study of O-GlcNAc moieties. In addition, these mAbs provide the first cytologic analysis of the distribution of O-GlcNAc residues throughout the nucleus and the cytoplasm of mammalian cells.

Acetylglucosamine↗

PubMed1990

Branched-chain fatty acids produced by mutants of Streptomyces fradiae, putative precursors of the lactone ring of tylosin.

Three branched-chain fatty acids (7-hydroxy-4,6-dimethylnona-2,4-dienoic acid [compound 1], its 7-epimer [compound 2], and 7-keto-4,6-dimethylnona-2,4-dienoic acid [compound 3]) and a ketone (9-hydroxy-6,8-dimethylundeca-4,6-dien-3-one [compound 4]) were isolated from the culture broth of mutants of Streptomyces fradiae which were blocked in the biosynthesis of the macrolide antibiotic tylosin. Two phenotypic classes of mutants of this organism which were blocked in the addition of mycaminose to tylactone (compound 6) accumulated these compounds. These compounds were not produced by mutants which were blocked in lactone synthesis, in steps beyond mycaminose addition, or by the wild-type strain. Synthesis of these compounds, like synthesis of tylosin, was inhibited by the addition of cerulenin. Compounds 1, 2, and 3 were partially interconvertible by these mutants; but they were not produced from the degradation of tylactone and they were not directly incorporated into tylosin by intact cells. The structures of compounds 1 and 2 were equivalent to that of a predicted intermediate (S. Yue, J. S. Duncan, Y. Yamamoto, and C. R. Hutchinson, J. Am. Chem. Soc. 109:1253-1255, 1987) in the biosynthesis of tylactone. The ketone (compound 4) reported previously (N. D. Jones, M. O. Chaney, H. A. Kirst, G. M. Wild, R. H. Baltz, R. L. Hamill, and J. W. Paschal, J. Antibiot. 35:420-425, 1982) appears to be the decarboxylation product of the intermediate following that represented by compound 1. This represents the first report of the isolation of putative precursors of tylactone from tylosin-producing organisms.

Alcohols↗

PubMed1990

Cloning of spiramycin biosynthetic genes and their use in constructing Streptomyces ambofaciens mutants defective in spiramycin biosynthesis.

Several cosmid clones from Streptomyces ambofaciens containing the spiramycin resistance gene srmB were introduced into S. fradiae PM73, a mutant defective in tylosin synthesis, resulting in tylosin synthesis. The DNA responsible for this complementation was localized to a 10.5-kilobase EcoRI fragment. A 32-kilobase DNA segment which included the srmB spiramycin resistance gene and DNA which complemented the defect in strain PM73 were mutagenized in vivo with Tn10 carrying the gene for Nmr (which is expressed in Streptomyces spp.) or in vitro by insertional mutagenesis with a drug resistance gene (Nmr) cassette. When these mutagenized DNA segments were crossed into the S. ambofaciens chromosome, three mutant classes blocked in spiramycin synthesis were obtained. One mutant accumulated two precursors of spiramycin, platenolide I and platenolide II. Two mutants, when cofermented with the platenolide-accumulating mutant, produced spiramycin. Tylactone supplementation of these two mutants resulted in the synthesis of a group of compounds exhibiting antibiotic activity. Two other mutants failed to coferment with any of the other mutants or to respond to tylactone supplementation.

Chromosomes, Bacterial↗

PubMed1990

Antibacterial properties of the bicyclic pyrazolidinones.

LY173013 and LY186826 are bicyclic pyrazolidinones containing a novel aza-gamma-lactam ring structure. The antibacterial properties of these compounds appear to be related to those of beta-lactam antibiotics in that both classes of compounds share certain common binding molecules such as beta-lactamases and penicillin-binding proteins.

Alanine↗

PubMed1990

On the relation between distinct components of the cytoskeleton: an epitope shared by intermediate filaments, microfilaments and cytoplasmic foci.

Monoclonal antibodies were generated against detergent-insoluble cytoskeletal proteins isolated from low-density membrane fractions of rat liver. By immunofluorescence, one of the antibodies stains three distinct structures in cultured rat fibroblast and hepatocyte lines as well as the PtK2 rat-kangaroo kidney epithelial line. These structures are: i) many tangled filaments similar to intermediate filaments (IFs), ii) fewer and variable numbers of straight filaments, and iii) punctate cytoplasmic foci, often most intense around the nucleus. All three of these structures are resistant to extraction by non-ionic detergent. Close examination reveals that the tangled and straight filaments are not stained uniformly, but as a series of bright patches. In cells treated with nocodazole, the antibody reacts strongly with a perinuclear filamentous cage. Very few tangled filaments are detected in these cells, however, the straight filaments and punctate cytoplasmic staining are resistant to nocodazole treatment. Double-label immunofluorescence shows that, even though tangled filament distribution and nocodazole sensitivity are similar to the behavior of vimentin IFs, there is only partial coincidence of staining with either vimentin or cytokeratin IFs. The straight filaments coincide with some actin stress fibers, but the punctate cytoplasmic staining is not related to IFs, actin, or tubulin. Thus, this monoclonal antibody stains a novel group of three seemingly unrelated cytoskeletal structures, including a previously undescribed insoluble nonfilamentous pool. Taken as a whole, two hypotheses are consistent with these data. i) The antigen recognized may be a protein which has a large insoluble cytoplasmic pool and binds both IFs and actin, but only binds to a subset of each class of filaments.(ABSTRACT TRUNCATED AT 250 WORDS)

Actin Cytoskeleton↗