Biomedical subjects

S White

Publications and source records attributed to S White.

At least 109 records · Page 6Linked to original sources

PubMed1999

Effect of phenylacetic acid feeding on the process of cellular autolysis in submerged batch cultures of Penicillium chrysogenum.

The effects of feeding the 'toxic' penicillin precursor, phenylacetic acid (PAA) at varying rates, upon the process of cellular autolysis, was assessed in batch bioreactor cultures of an industrial strain of Penicillium chrysogenum. Five processes were fed at rates which resulted in extracellular concentrations of PAA ranging from zero (the control) to approximately ten times levels said to be optimal for penicillin biosynthesis. The culture response was assessed chemically and morphologically, using computerised image analysis. High concentrations of PAA reduced biomass and penicillin production, and were associated with increased cellular autolysis. However, the values of classical morphological indices (branch length, main hyphal length and hyphal growth unit) varied little in cultures which showed extensive autolysis and biomass loss. Lower precursor concentrations (0.01 to 1.0 g l-1) had little effect on biomass, penicillin, or upon the levels of autolysis compared with the control process. Therefore, precursor concentration controlled within the optimal range for penicillin production, has little impact upon differentiation or degradation within an industrial culture of P. chrysogenum. By contrast, exploitation of the toxicity of PAA is proposed as a means to bring forward or enhance autolysis, providing a reliable method of 'induction' with which to study the phenomenon in P. chrysogenum.

Bacteriolysis↗

PubMed1999

Evidence for the head domain movement of the rieske iron-sulfur protein in electron transfer reaction of the cytochrome bc1 complex.

The three-dimensional structure of the mitochondrial cytochrome bc1 complex suggests that movement of the extramembrane domain (head) of the Rieske iron-sulfur protein (ISP) may play an important role in electron transfer. Such movement requires flexibility in the neck region of ISP, since the head and transmembrane domains of the protein are rather rigid. To test this hypothesis, Rhodobacter sphaeroides mutants expressing His-tagged cytochrome bc1 complexes with cysteine substitution at various positions in the ISP neck (residues 39-48) were generated and characterized. The mutants with a single cysteine substitution at Ala42 or Val44 and a double cysteine substitution at Val44 and Ala46 (VQA-CQC) or at Ala42 and Ala46 (ADVQA-CDVQC) have photosynthetic growth rates comparable with that of complement cells. Chromatophore membrane and intracytoplasmic membrane (ICM) prepared from these mutants have cytochrome bc1 complex activity similar to that in the complement membranes, indicating that flexibility of the neck region of ISP was not affected by these cysteine substitutions. Mutants with a double cysteine substitution at Ala42 and Val44 (ADV-CDC) or at Pro40 and Ala42 (PSA-CSC) have a retarded (50%) or no photosynthetic growth rate, respectively. The ADV-CDC or PSA-CSC mutant ICM contains 20 or 0% of the cytochrome bc1 complex activity found in the complement ICM. However, activity can be restored by the treatment with beta-mercaptoethanol (beta-ME). The restored activity is diminished upon removal of beta-ME but is retained if the beta-ME-treated membrane is treated with the sulfhydryl reagent N-ethylmaleimide or p-chloromercuribenzoic acid. These results indicate that the loss of bc1 complex activity in the ADV-CDC or PSA-CSC mutant membranes is due to disulfide bond formation, which increases the rigidity of ISP neck and, in turn, decreases the mobility of the head domain. Using the conditions developed for the isolation of His-tagged complement cytochrome bc1 complex, a two-subunit complex (cytochromes b and c1) is obtained from all of the double cysteine-substituted mutants. This suggests that introduction of two cysteines in the neck region of ISP weakens the interactions between cytochromes b, ISP, and subunit IV.

Base Sequence↗

PubMed1999

Microvessel precursor smooth muscle cells express head-inserted smooth muscle myosin heavy chain (SM-B) isoform in hyperoxic pulmonary hypertension.

The present study analyzes smooth muscle myosin heavy chain (SMMHC) expression as lung microvascular precursor smooth muscle cells (PSMCs), cells derived from fibroblasts and intermediate cells (immature SMCs), acquire a smooth muscle phenotype in an in vivo model of pulmonary hypertension (PH). Because of the unique contractile properties of the SMMHC isoform SM-B, we analyzed its expression in the microvessels (<100 micrometers diameter) and in larger vessels (100-700 micrometers) quantitatualy by the labeled [strept]avidin-biotin technique (day 1-28), and related this to cell phenotype by transmission microscopy and protein A-gold labeling (at day 28). Airway SMCs of the normal and hypertensive lung uniformly expressed SM-B whereas vascular SMC expression was heterogeneous. Thus, in some large arteries (and veins) SMCs contained cells expressing SM-B while in others all the cells were immunonegative. Microvascular cells expressing SM-B (arteries and veins) were rare in normal lung and numerous in PH, increasing as wall muscle developed in smaller segments with time. As in large vessels, some microvessels had immunopositive cells and others only negative ones. Ultrastructural analysis confirmed that the SMCs of bronchial vessels, and the septal SMCs adjoining alveolar ducts, contained dense filament arrays decorated with SM-B. While the PSMC processes of the normal lung contained sparse filaments decorated with SM-B, these cells expressed dense filament arrays in PH. Fibroblasts migrating to align around the microvessels also expressed SM-B but in the absence of a filament network. For the first time, we demonstrate in vivo that newly developed microvascular PSMCs express the SMMHC SM-B isoform in PH.

Animals↗

PubMed1999

Intermittent umbilical cord occlusion in the ovine fetus near term: effects on behavioral state activity.

OBJECTIVE: The purpose of this study was to determine the effects on fetal behavioral state activity of intermittent umbilical cord occlusion resulting in repetitive severe short-term hypoxemia. STUDY DESIGN: Fifteen near-term fetal sheep (experimental group, n = 8; control group, n = 7) were studied during 4 days while behavioral and cardiovascular parameters were monitored. Each day after a 2-hour control period, cord occlusions were performed in the experimental group animals by complete inflation of an occluder cuff (duration, 90 seconds) every 30 minutes for 3 to 5 hours. Results are presented as group mean +/- SEM. RESULTS: During umbilical cord occlusions fetal arterial PO(2) (change of 12 mm Hg), oxygen saturation (change of 40%), and glucose concentration (change of 0.3 mmol/L) fell and PCO(2) (change of 7 mm Hg) rose, but all returned toward control values after release of occlusion. Fetal behavioral state activity was markedly disrupted by 90 seconds of cord occlusion, with animals showing an abrupt flattening of the electrocorticogram. In >90% of instances the first identifiable state after cord release was the high-voltage non-rapid-eye-movement state. There was no apparent change in this response through the 4 days of the study. For experimental group animals the mean percentages of time spent in low-voltage electrocortical state (from 53 +/- 2 to 36 +/- 2), electro-ocular state (from 45 +/- 3 to 28 +/- 3), and fetal breathing activity (22 +/- 4 to 12 +/- 3) were significantly decreased (P <.001) during occlusion hours with respect to nonocclusion hours. CONCLUSION: Intermittent umbilical cord occlusion with severe but limited hypoxemia and no cumulative acidosis in the near-term ovine fetus disrupts behavioral state activity, with a flattening of the electrocortical activity during occlusions and an overall decrease in the prominence of the low-voltage rapid-eye-movement state. If such insults are frequent and severe enough, they might have an effect on growth and development of the brain during the perinatal period.

Animals↗

PubMed1999

Attitudes to oral feeding following caesarean section.

In order to evaluate postoperative nutrition in women who have undergone Caesarean section, we conducted a national survey. Questionnaires were sent to 100 randomly selected obstetric units in the UK, and were completed and returned by senior midwives. We found that that only 21.5% of units had a departmental policy concerning feeding after Caesarean section. Midwives decided when women could eat and drink in the majority of obstetric units (78.5%), often without the help of guidelines. The period of postoperative starvation was found to vary greatly, from < 1 h in some units to > 24 h in others. We suggest that all obstetric units should produce guidelines in order to rationalise postoperative feeding for women following Caesarean section.

Attitude of Health Personnel↗

PubMed1999

Using multiple response true-false multiple choice questions.

The Royal Australasian College of Surgeons (RACS) Board of Examiners, having agreed to replace Type K with Type X multiple choice questions (MCQ), undertook a study to determine the most appropriate method of scoring the latter. Retrospective scoring of candidates' responses in a previous examination, using six reported scoring techniques for Type X items, showed that the Middlesex technique yielded a mean score closest to the mean score of the Type K items which were being replaced. However, as candidates' responses are affected by instructions, and the latter could not be controlled, a prospective study was carried out on three examinations, in each of which the nature of Part C varied among the three papers, while Parts A and B were similar. Paper 1 contained Type K items in Part C, Paper 2 contained Type X items with instructions appropriate for the Middlesex technique, and Paper 3 contained Type X items scored +1 if correct, -1 if incorrect and 0 if omitted. Items sampling the same content areas were randomly distributed among the three papers. The results indicated that the two Type X systems yielded significantly lower mean scores than Type K, probably due to combinations characteristic of the latter, enabling candidates with partial knowledge to obtain a maximum score. The difference in mean scores between the two Type X systems was not significant. A significantly higher mean percentage score in reused Type X (Middlesex system) items over that of the same items when they were previously used as Type K items was attributed to the combined effects of credit for partial knowledge, practice on reused items and differences in mean candidate ability across years. Pearson product-moment correlation coefficients between pairs of Part C mean percentage scores at each examination were positive and significant. The Board decided to replace Type K with Type X items, and the +1, 0, -1 system of scoring was adopted as it was considered more efficient than the Middlesex system, which did not give credit for identifying false statements as false.

Australia↗

PubMed1999

Changes in serum chondroitin sulphate epitopes 3-B-3 and 7-D-4 in early rheumatoid arthritis.

OBJECTIVES: The aims of the present rheumatoid arthritis (RA) study were (1) to examine the levels of serum 3-B-3 and 7-D-4 to find out whether they are different from controls, (2) to find out whether the concentrations of these epitopes change with disease duration in early RA and (3) whether the serum concentrations of 3-B-3 and 7-D-4 in early RA are prognostic for subsequent disease progression. METHODS: The concentrations of 3-B-3 and 7-D-4 in sera were quantitated by immunoassays. RESULTS: The levels of 3-B-3 and 7-D-4 were significantly lower in RA than in controls (3- to 30-fold, P < 0.001). Changes in 3-B-3 and 7-D-4 were apparent with disease duration. At first presentation, the 3-B-3 concentration was lowest and increased at 12 months (3-fold, P < 0.001). This increase was transient since by 24 and 36 months the concentrations were not different to those at first presentation. The level of 7-D-4 was also lowest when the patients first presented at clinic and increased with time at 6 months (2-fold, P < 0.001). The increase was more prolonged for 7-D-4, remaining elevated at 12, 24 and 36 months. The lack of correlations of serum 3-B-3 and 7-D-4 with clinical measurements showed that these markers were not prognostic for disease severity. CONCLUSIONS: The levels of 3-B-3 and 7-D-4 differed between RA and control sera, and changed with disease duration. These markers were not prognostic in predicting disease outcome.

Adult↗

PubMed1999

Sweating and skin temperature responses of normal and anhidrotic horses to intravenous adrenaline.

Anhidrosis has been recognised for over half a century, but despite some excellent epidemiological studies, there has been little progress in understanding the aetiology of the condition. Using a modified ventilated capsule, we obtained dynamic, quantitative data on sweating responses in anhidrotic horses and normal sweating controls from the same environment. Ten horses with current seasonal anhidrosis and 10 matched normal sweating controls were selected. Each horse was given two 10 min infusions of 1 and 2 micrograms/kg/min adrenaline, separated by at least 6 h. Sweating responses and skin temperatures on the neck and gluteal region were measured. Plasma and sweat for analysis of total protein and electrolytes and plasma for analysis of adrenaline were collected. Anhidrotic horses produced significantly less sweat, had lower initial and peak sweat rates and a greater neck:gluteal ratio for sweat production. Plasma adrenaline at rest or at the time of peak sweating rate was not different between groups. In nearly three-quarters of the anhidrotic horses, the shape of the sweat rate against time curve was different compared to controls. Volume of sweat produced was significantly correlated with skin temperature on the neck of controls and anhidrotic horses and on the gluteal region of controls, but not anhidrotic horses. Plasma total protein and electrolyte concentrations were not different between groups. There were significant differences in sweat electrolyte concentrations between controls and anhidrotic horses. These differences were reduced when sweat electrolytes were expressed per g of total protein, and no differences existed when expressed as g/m2. This study has provided insight into the response of anhidrotic horses to beta 2 adrenergic stimulation and may be a useful technique to investigate this condition.

Animals↗

PubMed1999

The oral contraceptive pill: a revolution for sportswomen?

OBJECTIVES: To determine the effects of the oral contraceptive pill (OCP) on skeletal health, soft tissue injury, and performance in female athletes. METHODS: A literature review was performed using literature retrieval methods to locate relevant studies. RESULTS: Most female athletes primarily choose to use the OCP for contraceptive purposes, but cycle manipulation and control of premenstrual symptoms are secondary advantages of its use. The effect of the OCP on bone density in normally menstruating women is unclear, with some studies reporting no effect, others a positive effect, and some even a negative effect. The OCP is often prescribed for the treatment of menstrual disturbances in female athletes, and improvements in bone density may result. Whether the OCP influences the risk of stress fracture and soft tissue injuries is not clear from research to date. Effects of the OCP on performance are particularly relevant for elite sportswomen. Although a reduction in Vo2MAX has been reported in some studies, this may not necessarily translate to impaired performance in the field. Moreover, some studies claim that the OCP may well enhance performance by reducing premenstrual symptoms and menstrual blood loss. A fear of weight gain with the use of the OCP is not well founded, as population studies report no effect on weight, particularly with the lower dose pills currently available. CONCLUSIONS: Overall, the advantages of the pill for sportswomen would appear to outweigh any potential disadvantages. Nevertheless, there is individual variation in response to the OCP and these should be taken into account and monitored in the clinical situation. Women should be counselled as to the range of potential benefits and disadvantages in order to make an informed decision based on individual circumstances.

Athletic Injuries↗

PubMed1999

Social Security fixes--beware of conjurers!

Among the many proposals for reform, there is one that offers the best chance of truly solving Social Security's financial problems, these authors believe. That proposal would increase savings by requiring workers to invest an additional 2% of their covered wages in individual accounts. At retirement, 75% of the money would go toward buying the current level of Social Security benefits, and 25% would be given to the individual as an "extra" pension. Individuals, not the government, would control investment of these accounts.

Actuarial Analysis↗

PubMed1999

Improved percutaneous slipped capital femoral epiphysis treatment: continuous biplanar fluoroscopy and proper guide wire selection.

In 1990, a report was published outlining a technique of percutaneous fixation of slipped capital femoral epiphyses dramatically diminishing the morbidity associated with the open technique. Technical difficulties are still encountered with the morbidly obese child and the percutaneous technique. Two fluoroscopic units used continuously during the technique facilitate placement of the guide wire in a more acceptable anatomic location. Guide wire stiffness was also measured and used in preoperative planning. Ten hips were treated with this technique, and a prospective analysis of the surgical time and fluoroscopy time was done. Using the new technique, we found a significant reduction in surgical and fluoroscopic times.

Bone Screws↗

PubMed1999

My good friend Roe.

Explore the source record for details and available documents.

Abortion, Induced↗

PubMed1998

A site for CD4 binding in the beta 1 domain of the MHC class II protein HLA-DR1.

Using a lymphocyte binding assay, we have previously demonstrated that the CD4 protein can mediate cell adhesion by direct interaction with MHC class II molecules. In this report, we have used this assay to test whether synthetic peptides, corresponding to DR beta sequences, could inhibit CD4-class II adhesion. A peptide derived from sequences within the beta1 domain (DR beta 41-55), as well as two peptides derived from sequences within the beta 2 domain (DR beta 121-135 and DR beta 141-155), were shown to inhibit CD4-class II adhesion. Inasmuch as a site for CD4 binding in the beta 2 domain had been previously documented, these studies were designed to investigate the role of the beta 1 domain as an additional site of interaction with CD4. Sixteen site-specific mutations were engineered within the beta1 domain of DR beta 1*0101. Several mutations were shown to disrupt CD4-dependent T cell activation. Based on these results, we propose a model for the molecular interaction of CD4 with MHC class II proteins in which both the beta 1 and beta 2 domains of class II interact with the two amino-terminal Ig-like domains of CD4.

Amino Acid Sequence↗

PubMed1998

A structural basis for recognition of A.T and T.A base pairs in the minor groove of B-DNA.

Polyamide dimers containing three types of aromatic rings-pyrrole, imidazole, and hydroxypyrrole-afford a small-molecule recognition code that discriminates among all four Watson-Crick base pairs in the minor groove. The crystal structure of a specific polyamide dimer-DNA complex establishes the structural basis for distinguishing T.A from A.T base pairs. Specificity for the T.A base pair is achieved by means of distinct hydrogen bonds between pairs of substituted pyrroles on the ligand and the O2 of thymine and N3 of adenine. In addition, shape-selective recognition of an asymmetric cleft between the thymine-O2 and the adenine-C2 was observed. Although hitherto similarities among the base pairs in the minor groove have been emphasized, the structure illustrates differences that allow specific minor groove recognition.

Adenine↗

PubMed1998

Analysis of the structural requirements for lysosomal membrane targeting using transferrin receptor chimeras.

The sorting of membrane proteins to the lysosome requires tyrosine- or dileucine-based targeting signals. Recycling receptors have similar signals, yet these proteins seldom enter the latter stages of the endocytic pathway. To determine how lysosomal and internalization signals differ, we prepared chimeric molecules consisting of the cytoplasmic tails of CD3 gamma-chain, lysosomal acid phosphatase, and lysosomal-associated membrane glycoprotein-1, each fused to the transmembrane and extracellular domains of the transferrin receptor (TR). Each chimera was expressed on the cell surface and rapidly internalized. Metabolic pulse-chase experiments showed that the CD3 gamma-chain and lysosomal acid phosphatase chimeras, unlike the lysosomal-associated membrane glycoprotein chimera, were rapidly degraded in a post-Golgi compartment following normal glycosylation. Transplantation of signals from CD3 gamma-chain and lysosomal acid phosphatase into the TR cytoplasmic tail in place of the native signal, Y20TRF23, indicated that each signal was sufficient to promote endocytosis but not lysosomal targeting of the resulting mutant. Transplantation of two CD3 signals at specific sites in the TR cytoplasmic tail or a single tyrosine-based signal in a truncated TR tail, however, was sufficient to promote lysosomal targeting. Our results therefore suggest that the relative position of the signal within the cytoplasmic tail is a critical feature that distinguishes lysosomal targeting signals from internalization signals.

Acid Phosphatase↗

PubMed1998

The reduction potential of cytochrome b5 is modulated by its exposed heme edge.

When the reduction potential of cytochrome b5 is measured with the aid of several different surface-modified electrodes that function on the basis of electrostatic interactions with the protein, the resultant values have been consistently more positive (40-100 mV) than the reduction potentials measured with potentiometric methods. In this paper, we report that the heme edge containing the exposed heme propionate, a heme methyl, and a heme vinyl, and which constitutes part of the surface of cytochrome b5, modulates its reduction potential. The positive shifts observed in the voltammetric measurements appear to originate from the formation of a complex between cytochrome b5 and the modified electrode surface which (a) neutralizes the charge on the heme propionate located on the exposed heme edge and (b) lowers the dielectric of the exposed heme microenvironment by excluding water from the complex interface, factors which result in the destabilization of the positive charge on the ferric heme with respect to the neutral ferrous heme. The observed positive shift, which is induced by complexation at the electrode surface, may indicate that similar shifts in the reduction potential of cytochrome b5 occur when it forms a complex with physiological partners, prior to electron transfer. The effect of the value of the dielectric constant on the reduction potential of cytochrome b5 was corroborated by preparing the V45L/V61L double mutant whose reduction potential was measured to be 50 mV more negative than the value measured for the wild type protein. The negative shift in the reduction potential of the mutant protein was explained by the increased accessibility of water to the heme binding site, as observed in its X-ray crystal structure.

Amino Acid Substitution↗

PubMed1998

Measurement of autolysis in submerged batch cultures of Penicillium chrysogenum.

The process of cellular autolysis was studied in an industrial strain of Penicillium chrysogenum by a range of methods, including assessment of biomass decline, NH+4 release, changes in culture apparent viscosity, and by means of a quantitative assessment of changes in micromorphology using a computerized image analysis system. The pattern of total intracellular proteolytic and beta-1, 3-glucanolytic activity in the culture was also examined. The overall aim was to identify a suitable method, or methods, for examining the extent of autolysis in fungal cultures. Autolysis was studied in submerged batch processes, where DOT was maintained above 40% saturation (non-O2-limited), and, under O2-limited conditions. Both N and O2 limitation promoted extensive culture autolysis. Image analysis techniques were perhaps the most sensitive method of assessing the progress of autolysis in the culture. Autolytic regions within some hyphae were apparent even during growth phase, but became much more widespread as the process proceeded. The early stages of autolysis involved continued energy source consumption, increased carbon dioxide evolution rate, degradation of penicillin, and decreased broth filterability. Later stages involved widespread mycelial fragmentation, with some regrowth (cryptic growth) occurring in non-O2-limited cultures. Intracellular proteolytic activity showed two peaks, one during the growth phase, and the other during autolysis. Autolysis was also associated with a distinct peak in beta-1,3-glucanolytic activity, indicating that degradation of cell wall matrix polymers may be occurring during autolysis in this strain of P. chrysogenum.

Biomass↗

PubMed1998

Recognition of the four Watson-Crick base pairs in the DNA minor groove by synthetic ligands.

The design of synthetic ligands that read the information stored in the DNA double helix has been a long-standing goal at the interface of chemistry and biology. Cell-permeable small molecules that target predetermined DNA sequences offer a potential approach for the regulation of gene expression. Oligodeoxynucleotides that recognize the major groove of double-helical DNA via triple-helix formation bind to a broad range of sequences with high affinity and specificity. Although oligonucleotides and their analogues have been shown to interfere with gene expression, the triple-helix approach is limited to recognition of purines and suffers from poor cellular uptake. The subsequent development of pairing rules for minor-groove binding polyamides containing pyrrole (Py) and imidazole (Im) amino acids offers a second code to control sequence specificity. An Im/Py pair distinguishes G x C from C x G and both of these from A x T/T x A base pairs. A Py/Py pair specifies A,T from G,C but does not distinguish AT from T x A. To break this degeneracy, we have added a new aromatic amino acid, 3-hydroxypyrrole (Hp), to the repertoire to test for pairings that discriminate A x T from T x A. We find that replacement of a single hydrogen atom with a hydroxy group in a Hp/Py pairing regulates affinity and specificity by an order of magnitude. By incorporation of this third amino acid, hydroxypyrrole-imidazole-pyrrole polyamides form four ring-pairings (Im/Py, Py/Im, Hp/Py and Py/Hp) which distinguish all four Watson-Crick base pairs in the minor groove of DNA.

Amino Acids↗