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Biomedical subjects

D R Jones

Publications and source records attributed to D R Jones.

At least 91 records · Page 5Linked to original sources
PubMed1999

Reduced ischemia-reperfusion injury with rolipram in rat cadaver lung donors: effect of cyclic adenosine monophosphate.

BACKGROUND: The perfusion of rat lungs retrieved from cadavers with a solution containing isoproterenol has been shown to ameliorate the ischemia-reperfusion injury seen in lungs retrieved after death, and this protective effect parallels increases in tissue cyclic adenosine monophosphate levels. In this study, we investigated the effect of rolipram, a phosphodiesterase inhibitor, on capillary permeability and lung cyclic adenosine monophosphate levels in lungs retrieved from circulation-arrested rats. METHODS: Using an isolated perfused lung circuit, we retrieved lungs from circulation-arrested donor rats either ventilated with 100% oxygen or not ventilated for varying postmortem times. The lungs were reperfused with or without rolipram (2 micromol/L). The capillary filtration coefficient and wet to dry weight ratio, indicators of pulmonary vascular integrity, were determined, and tissue levels of adenine nucleotides and cyclic adenosine monophosphate were measured by high-performance liquid chromatography. RESULTS: The capillary filtration coefficient was significantly reduced in nonventilated cadaver lungs reperfused with rolipram 120 minutes after death (p<0.05). Oxygen ventilation or reperfusion with rolipram had a similar effect on the capillary filtration coefficient. Cyclic adenosine monophosphate levels were significantly higher in rolipram-reperfused lungs retrieved 120 minutes after death in both oxygen-ventilated (p<0.01) and nonventilated (p<0.01) lungs. CONCLUSIONS: In lungs from nonventilated, circulation-arrested donors, reperfusion with rolipram reduces the ischemia-reperfusion injury that may be due to intracellular cyclic adenosine monophosphate. Alteration of perfusate may have an impact on capillary leak caused by antecedent ischemia. Thus, rolipram may be a useful adjunct in the preservation of donor lungs retrieved after death.

Adenine Nucleotides↗
PubMed1999

Co-cultivation of CD4+ and CD8+ human T-cells leads to the appearance of CD4 cells expressing CD8 through de novo synthesis of the CD8 alpha-subunit.

The appearance of a population of dual-staining CD4+CD8+ cells in human T-lymphocyte cultures has been reported by various authors, including our own observation that they are always seen in simple phytohaemagglutinin-stimulated cultures from several different donors. The purpose of the present study was to investigate factors involved in the dual-staining (DS) phenotype, and to clarify some apparent inconsistencies between published observations. Our findings can be summarised as follows. 1. A population of DS CD4+CD8+ cells always appears in PHA-stimulated T-cell cultures if they contain both CD4 and CD8 subsets. The incidence of DS cells is related to PHA concentration, but other factors are involved since DS cells are not seen in PHA-stimulated cultures of purified CD4+ or CD8+ cells. Stimulation with PHA is not a prerequisite since very similar results are seen with ConA. 2. Direct physical contact between CD4+ and CD8+ cells is required for the appearance of the DS phenotype; soluble factors alone, including IL-4, appear nor to be responsible. 3. The DS phenotype in these conditions is always CD4+ cells weakly expressing CD8 and is a consequence of de novo synthesis of the CD8alpha molecule by the CD4+ cells.

Apoptosis↗
PubMed1999

Cardiac responses to first ever submergence in double-crested cormorant chicks (Phalacrocorax auritus).

Heart rates were recorded from double-crested cormorant chicks during their first ever and subsequent voluntary head submergences and dives, as well as during longer dives made after the chicks were accustomed to diving. Despite variation between chicks, the cardiac response to first ever and subsequent voluntary submergence (head submergences and dives) was similar to the response observed in adult cormorants. Upon submersion the heart rate fell rapidly when pre-submersion heart rate was high (325-350 beats min-1). The heart rate established within the first second of voluntary submergence was between 230 and 285 beats min-1, well above resting heart rate (143 beats min-1). The same initial cardiac response occurred during longer dives performed after the chicks were accustomed to diving. In these dives the heart rate remained at the level established on submersion, unlike the response observed in shallow diving adult cormorants in which the heart rate declined throughout the dive. The heart rate was also monitored in a separate group of chicks in which the first exposure to water was during whole body forced submergence. Again, the observed response was similar to the adult response, although the cardiac response of chicks to forced submergence was more extreme than to voluntary submergence. Our results do not support the hypothesis that learning (by conditioning or habituation) is involved in the cardiac adjustments to voluntary submergence. It is suggested that the initial cardiac adjustments are reflex in nature and this reflex is fully developed by the first submergence event. Although the nature of this reflex pathway is obscure, cessation of breathing before submersion and the close linkage between breathing and heart rate might provide a plausible mechanism.

Age Factors↗
PubMed1999

Why do in utero stem cell transplants sometimes fail?

In utero stem cell transplantation promises a novel therapeutic approach to those genetic disorders that can be diagnosed in early pregnancy and that could lead to either severe disability or death. Available scientific evidence suggests that such procedures could achieve clinically relevant levels of engraftment with donor cells and that the resulting sustained chimerism is potentially long lived. However, the relatively few cases performed so far have not borne out initial hopes, and both the source and the type of cell preparation to be transplanted and the choice of disorders to target with this therapy remain controversial.

Animals↗
PubMed1999

Calcium-independent haemolysis via the lectin pathway of complement activation in the guinea-pig and other species*.

We previously reported that complement-dependent haemolysis of sheep erythrocytes (E) coated with mannan (M) and sensitized with human mannan-binding lectin (MBL) via the lectin pathway in man occurs in Mg-EGTA and requires alternative pathway amplification. Calcium was required for MBL binding to E-M, but once the E-M-MBL intermediate was formed, MBL was retained and haemolysis occurred in the absence of calcium. Comparable or greater lectin pathway haemolysis in the absence of calcium was observed upon incubation of E-M-MBL in guinea-pig, rat, dog and pig sera, and was further investigated in the guinea-pig, in which titres were much higher ( approximately 14-fold) than in man, and in contrast to humans, greater than classical pathway haemolytic activity. As in human serum, no lysis was observed in C4- or C2-deficient guinea-pig serum until purified C4 or C2, respectively, were restored. However, lectin pathway haemolytic activity in the guinea-pig did not require the alternative pathway. Removal (>98%) of factor D activity by three sequential passages through Sephadex G-75, resulting in serum which retained a normal classical pathway but no alternative pathway haemolytic activity, did not reduce the ability of guinea-pig serum to mediate haemolysis via the lectin pathway. Further, the C3-convertase formed via the lectin pathway (E-M-MBL-C4,2) lysed in C2-deficient guinea-pig but not human serum chelated with EDTA, a condition which precludes alternative pathway amplification. Thus, lectin pathway haemolysis occurs efficiently in guinea-pig serum, in the absence of calcium and without requirement for alternative pathway amplification. The guinea-pig provides a model for studying the assembly and haemolytic function of a lectin pathway which contrasts with the lectin pathway of man, and allows for comparisons that may help clarify the role of this pathway in complement biology.

Animals↗
PubMed1999

Biotransformation of alprazolam by members of the human cytochrome P4503A subfamily.

1. To aid in the prediction of drug interactions with alprazolam, the human CYP involved in the 1'- and 4-hydroxylation of alprazolam were characterized using human liver microsomes, expressed enzymes and selective chemical inhibitors. 2. The formation of 4-hydroxyalprazolam and 1'-hydroxyalprazolam at an alprazolam concentration of 62.5 microM were reduced by the prototypic CYP3A inhibitor, troleandomycin (50 microM), by 97 and 9900 respectively. Only microsomes from B-lymphoblastoid cells expressing CYP3A4 were capable of catalysing the 1'- and 4-hydroxylation of alprazolam. 3. The formation rates of 1'-hydroxyalprazolam and 4-hydroxyalprazolam at an alprazolam concentration of 1 mM were significantly correlated (n = 19, r = 0.95, p<0.01) indicating that the same enzyme(s) mediated these biotransformations. A significant (p<0.01) correlation was observed between alprazolam 4- and 1'-hydroxylase activity and CYP3A-mediated midazolam 4-hydroxylase, midazolam 1'-hydroxylase, dextromethorphan N-demethylase and erythromycin N-demethylase activities. 4. In conclusion, in adult human liver the CYP3A subfamily members are the principal enzymes involved in the 1'- and 4-hydroxylation of alprazolam. Thus, clinically significant drug drug interactions between alprazolam and other CYP3A substrates are to be expected.

Alprazolam↗
PubMed1999

A bifunctional regulatory element of the human ApoA-I gene responsive to a distal enhancer.

Promoter elements located up to 2 kb upstream of the apolipoprotein A-I (apoA-I) gene are necessary for apoA-I expression in liver and intestine cells in tissue culture. In transgenic mice, a distal enhancer located between the apoA-IV and apoC-III genes is additionally necessary for tissue-specific expression of apoA-I in liver and intestine. We have identified a previously uncharacterized regulatory element between 746 and 856 nucleotides 5' of the apoA-I transcription start site that differentially affects the expression of apoA-I reporter plasmids in intestine cells dependent on the presence of the distal apolipoprotein enhancer. Deletion of the -856/-746 sequence strongly repressed transcription in the presence of the apolipoprotein enhancer, but in the absence of the enhancer, deletion of the -856/-746 element increased transcription. By contrast, in liver cells, deletion of the -856/-746 element strongly repressed transcription in the presence of the distal enhancer but had no detectable effect on transcription in the absence of the distal enhancer. Electrophoretic mobility shift analysis revealed tissue-specific and sequence-specific protein-DNA complexes formed by the -856/-746 element in intestine, liver, and HeLa cell nuclear extracts. The complexes formed by extracts of intestinal cells differed from those of liver and HeLa cells by their sensitivity to DNase digestion and their pattern of protein footprints. Collectively, the data suggest that the -856/-746 sequence is a composite regulatory element that interacts with multiple proteins and the apolipoprotein distal enhancer to achieve tissue-specific expression of apoA-I.

Apolipoprotein A-I↗
PubMed1999

Conservative management of splenic trauma.

conservative approach to splenic trauma has been practised in many countries. Haemodynamically stable patients who have been carefully assessed clinically and radiographically may safely be treated non-operatively. In those patients who require surgery the spleen may be preserved by splenorrhaphy or partial splenectomy. This approach has been practised at our hospital and we present our experience over seven years to show that expectant treatment of splenic injury following trauma is safe.

Accidents, Traffic↗
PubMed1999

Systematic reviews and meta-analysis: a structured review of the methodological literature.

OBJECTIVE: To systematically review methods for systematic review/meta-analysis in order to identify the different methodological and statistical methods that have been proposed. A summary of the main findings is presented here, with emphasis given to health services research topics. METHODS: A thorough systematic search for methodological papers was carried out using a variety of methods, including the use of electronic databases. Approximately 1000 potentially relevant references were identified, a number of them from education, psychology and sociology. RESULTS: After briefly reviewing the procedural methods required to carry out a review, and the basic statistical methods used to combine study estimates, less established methods are discussed. These include methods for dealing with publication bias, meta-regression, meta-analysis of individual patient data, the synthesis of non-randomized evidence alone and in combination with randomized studies. Bayesian modelling and economic evaluation through meta-analysis. Recommendations for meta-analytical practice are given; these are either distilled from previous guidelines, or constructed where there appears to be a broad consensus across the literature. CONCLUSIONS: It is hoped that this review will provide a consistent and comprehensive, but concise, description of the methods available for synthesizing evidence, that it will promote better quality reviews of the results of health services research and identify specific areas which require methodological development.

Bayes Theorem↗
PubMed1999

Vibroacoustic disease: some forensic aspects.

BACKGROUND: Vibroacoustic disease (VAD) is an insidious environmental entity caused by occupational exposure to large pressure amplitude and low frequency (LPALF) noise (> or = 90 dB SPL, < or = 500 Hz). Significant disabilities may result, and issues of worker's compensation should be openly discussed and settled. Toward this goal, a rigorous review of all available information on this disease was undertaken in order to document the need to define industrial exposure standards, and to settle on the diagnostic procedures that will help distinguish the VAD from other, similar conditions. METHODS: We reviewed the medical files of 236 male Caucasians employed as aircraft technicians who had been diagnosed with VAD and monitored for at least 15 yr. The natural history of the disorder was studied in detail, emphasizing the clinical diagnostic criteria and the outcome of disabilities. RESULTS: Among the 236 cases, 172 (73%) were disabled after an average of 24 yr (SD = 6.9) of occupational exposure. The primary categories of disabilities were neurological (81, 34%), malignant (28, 11.9%), psychiatric (23, 9.7%), cardiovascular (16, 6.8%), and osteoarticular (14, 5.9%). After the onset of industrial exposure to LPALF noise, the minimum time for these disabilities to manifest themselves was 16 yr. The most serious complication was multiple attempted suicide. Such attempts were fortunately rare (5, 2.1%) and successful only once. Echocardiograms showed characteristic changes in pericardial structures, involving proliferation of the extracellular matrix, lack of cilia and five pericardial layers instead of three. This has been studied in pericardial biopsy material obtained during coronary bypass surgery for coronary insufficiency (six cases), and by autopsy (four cases). DISCUSSION: These findings appear to be pathognomonic for the VAD, and the echocardiogram has been confirmed as a fundamental diagnostic tool. The degree of disability due to VAD can be determined from Portuguese national disability tables, which cover almost all of the VAD-induced disabilities. However, these tables do not specify LPALF noise as an occupational hazard, rendering them inapplicable to VAD patients. Also, suicide, a most serious psychiatric consequence, is not covered by these tables. Such situations highlight the absolute necessity to recognize LPALF noise as the cause of VAD, and as an industrial hazard.

Adult↗
PubMed1999

Diltiazem inhibition of cytochrome P-450 3A activity is due to metabolite intermediate complex formation.

Diltiazem (DTZ) N-demethylation occurs by cytochrome P-450 (CYP) 3A based on the following observations: 1) a single enzyme Michaelis-Menten model of metabolite formation, 2) high correlations of DTZ N-demethylation activity to other CYP3A activities, 3) inhibition of DTZ N-demethylation activity by triacetyloleandomycin, and 4) DTZ N-demethylation activity by expressed CYP3A enzymes only. The mean K(m)s for DTZ N-demethylation in human liver microsomes and expressed CYP3A4(+b(5)) were 53 and 16 microM, respectively. A 30-min preincubation of DTZ in expressed CYPs inhibited CYP3A4(+b(5)) by 100%, of which 55% was due to formation of a metabolite intermediate complex (MIC), which is an inactive form of CYP. MIC was observed in human liver microsomes and cDNA-expressed CYP3A only. In experiments to assess simultaneous MIC formation and loss of CYP3A activity, DTZ caused greater than 80% inhibition of midazolam hydroxylation after a 60-min preincubation in human liver microsomes. The rate constants for MIC formation and loss of midazolam hydroxylation activity were equivalent for the line of best fit for both data sets, which illustrates that MIC formation causes the inhibition of CYP3A activity. The mechanistic inhibition was characterized in expressed CYP3A4(+b(5)), which exhibited a concentration-dependent formation of MIC by DTZ (1-100 microM) with an estimated k(inact) of 0.17 min(-1) and K(I) of 2.2 microM. The partition ratio for expressed CYP3A4(+b(5)) was substrate concentration dependent and varied from 13 to 86. This study showed that DTZ inhibition of CYP3A substrate metabolism occurs primarily by MIC formation.

Adult↗
PubMed1999

Diabetes and the role of inositol-containing lipids in insulin signaling.

Among metabolic diseases, diabetes is considered one of the most prevalent throughout the world. Currently, statistics show that over 10% of the world's aged population (60 years and older) suffers from diabetes. As a consequence, it consumes a considerable proportion of world health expenditure. This review considers both past and current research into the molecular basis of insulin resistance found in type II diabetes and focuses on the role of inositol-containing phospholipid metabolism. It has been firmly established that the activation of phosphatidylinositol 3-kinase (PI3-K) is important for the propagation of the metabolic actions of insulin. In addition to the 3-phosphorylated phosphatidylinositols formed via the action of PI3-K, the glycosyl-phosphatidylinositol/inositol phosphoglycan (GPI/IPG) signaling component is also strongly implicated in mediating numerous metabolic actions of insulin. Although all the elements within the type II diabetes phenotype have not been fully defined, it has been proposed that defects in insulin transmembrane signaling through malfunction of inositol-containing phospholipid metabolism and absenteeism of the generation of phospholipid-derived second messengers may be associated with the appearance of the type II diabetic phenotype. Pharmaceutical approaches using synthetically produced IPG analogues, which themselves mimic insulin's actions, alone or in combination with other drugs, may lead the way toward introducing alternative therapies for type II diabetes in the coming years.

Aged↗
PubMed1998

Interleukin-2 stimulates a late increase in phosphatidic acid production in the absence of phospholipase D activation.

The signal transduction pathways involving phospholipid metabolism during T-cell proliferation remain partly undefined. Herein we show that interleukin-2 caused a late (> 12 h) rise in the intracellular phosphatidic acid content of CTLL-2 cells which was a consequence of the activation of the enzyme diacylglycerol kinase. No activation of phospholipase D was observed at similar times. Incubation of the cells with a recognized diacylglycerol kinase a isoform inhibitor, R59499, prior to interleukin-2 stimulation was able to block cell cycle entry, diacyglycerol kinase activation and phosphatidic acid accumulation. In contrast, when R59499 was added 3 h after interleukin-2, few or no observable effects on the above three parameters were noticed. These results suggest that the early signaling employed by IL-2 involving the alpha isoform of diacylglycerol kinase is sufficient to control the late increase in phosphatidic acid and that phosphatidic acid is a mitogenic agent in T-cells.

3T3 Cells↗
PubMed1998

Phospholipase cleavage of glycosylphosphatidylinositol reconstituted in liposomal membranes.

Glycosylphosphatidylinositol (GPI) purified from rat liver lipids was incorporated into lipid bilayers of defined compositions, in the form of large unilamellar vesicles. The GPI concentration in the bilayers was kept constant at 25 mole%, whereas the remaining lipids being phosphatidylcholine, phosphastidylethanolamine, sphingomyelin and/or cholesterol were varied. The resulting liposomes consisted of spherical vesicles, approximately 100 nm in diameter, that could keep their aqueous contents separated from the extravesicular medium. When these liposomes were treated with either Bacillus cereus phosphatidylinositol-phospholipase C, Trypanosoma brucei GPI-phospholipase C, or bovine serum GPI-phospholipase D, GPI was hydrolyzed at different rates, depending on the enzyme and the bilayer lipid composition. These observations open the way to biophysical and biochemical studies of enzymic GPI cleavage under defined conditions. Extensive GPI hydrolysis was observed in certain cases that could allow the use of these systems for the preparation of inositol phosphoglycans, proposed second messengers of a wide variety of hormones, cytokines and growth factors.

Animals↗