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Biomedical subjects

B J Clark

Publications and source records attributed to B J Clark.

At least 145 records · Page 8Linked to original sources

Computer-aided optimisation of drug enantiomer separation in chiral high-performance liquid chromatography.

The advent of several new column materials for the resolution of chiral compounds in high-performance liquid chromatography has opened up new possibilities for the analysis of drug enantiomers both in the dosage form and in bioanalytical studies. The utility of simplex optimisation, modified simplex and response surface mapping are considered with reference to the antischistosomal drug, oxamniquine, separated on an alpha 1-acid glycoprotein column. The resolution of the enantiomers of three closely related benzodiazepines, temazepam, oxazepam and lorazepam, is attempted on three new column systems: cellulose triacetate, beta-cyclodextrin and the reversed-phase column porous graphitic carbon with beta-cyclodextrin as a mobile phase additive.

Chromatography, High Pressure Liquid↗

Digital techniques for luminescence detection in liquid chromatography with an intensified linear photodiode array.

The use of a rapid-scanning fluorescence detector in liquid chromatography is examined in the context of its potential contribution in pharmaceutical and biomedical analysis. The data generated by an intensified linear photodiode array detector is presented as an isometric plot of (I(em),lambda(em),t) at a defined excitation wavelength. Techniques examined for peak homogeneity assessment include: emission spectral normalisation at points through the chromatographic peak profile, second order differentiation (d(2)I/dt(2)) and the fluorescence emission ratio chromatogram, generated by calculating the ratio of emission intensifies at two defined emission wavelengths, at all points in the time domain elution profile. These techniques are illustrated with reference to some polynuclear aromatic hydrocarbons and to the beta-blocker drug atenolol and its related impurities. Future developments of this new detector technology in LC are also considered.

Journal Article↗

In vivo 31P-NMR spectroscopy of chronically stimulated canine skeletal muscle.

Chronic stimulation converts skeletal muscle of mixed fiber type to a uniform muscle made up of type I, fatigue-resistant fibers. Here, the bioenergetic correlates of fatigue resistance in conditioned canine latissimus dorsi are assessed with in vivo phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy. After chronic electrical stimulation, five dogs underwent 31P-NMR spectroscopic and isometric tension measurements on conditioned and contralateral control muscle during stimulation for 200, 300, 500, and 800 ms of an 1,100-ms duty cycle. With stimulation, phosphocreatine (PCr) fell proportional to the degree of stimulation in both conditioned and control muscle but fell significantly less in conditioned muscle at all but the least intense stimulation period (200 ms). Isometric tension, expressed as a tension time index per gram muscle, was significantly greater in the conditioned muscle at the two longest stimulation periods. The overall small change in PCr and the lack of a plateau in tension observed in the conditioned muscle are similar to that seen in cardiac muscle during increased energy demand. This study indicates that the conditioned muscle's markedly enhanced resistance to fatigue is in part the result of its increased capacity for oxidative phosphorylation.

Animals↗

Pediatric chest pain: a prospective study.

Previous studies of childhood chest pain have been retrospective or considered only limited age groups or referred patients. In this study, all children who were admitted to the emergency department with chest pain were evaluated prospectively. Patients with ill-defined chest pain had ECGs and echocardiograms performed. A total of 407 children were evaluated. The most common causes of the pain were idiopathic (21%) and musculoskeletal (15%). Cardiac problems were found in 4%. Chest pain was acute (of less than 48 hours' duration) in 43% and chronic (of greater than 6 months' duration) in 7%. Pain caused 30% of children to stay out of school and 31% to awaken from sleep. Chest wall tenderness was the most common abnormality. ECGs were obtained in 47%; results of 31/191 were abnormal but only 4/191 ECG abnormalities were related to the diagnosis. Echocardiograms were obtained in 34%; results of 17/139 were abnormal (12/139 showed mitral valve prolapse). Young children are more likely to have cardiorespiratory problems; children older than 12 years of age are more likely to have psychogenic pain. The description and location of the pain and the patient's sex are not related to the diagnosis. Nonorganic disease is related to a family history of heart disease or chest pain or having chronic pain. Organic disease is related to pain of acute onset, abnormal physical examination results, pain that awakens the child from sleep, and the presence of fever. Laboratory tests are rarely helpful in evaluating children with chest pain. Chest pain in children is usually benign. Psychogenic pain and idiopathic pain are less common than previously believed.

Acute Disease↗

A new semisynthetic ergot peptide alkaloid: DCN 203-922.

We report the synthesis, stereochemistry and preliminary pharmacological evaluation of DCN 203-922, a novel ergot alkaloid of the cyclol type, which contains in its peptide moiety the uncommon amino acid L-allo-isoleucine.

Animals↗

Cardiac transfer function relating energy metabolism to workload in different species as studied with 31P NMR.

Cardiac metabolism was studied with 31P NMR in 7 dogs and 4 cats to determine whether animals adapted for different life-styles (stalk and sprint vs endurance running) respond to increased work loads (heart rate X blood pressure product) with different high-energy phosphate kinetics. Hearts were exposed via a left lateral thoracotomy under Nembutal anesthesia (40 mg/kg). Two-turned solenoid surface coils were placed on the left ventricles; pacing wires were sutured into the left ventricular apices. The femoral artery and vein were cannulated for blood pressure and arterial blood gas monitoring and fluid and drug infusion, respectively. Animals were placed in a plexiglass holder into a 2.1-T, 31-cm-bore, superconducting magnet. 31P spectra were obtained from the heart using respiratory and electrocardiogram gating. Cardiac work loads were changed by pacing the heart at 4, 4.5, and 5 Hz. Heart rate X blood pressure product "work" was correlated with Pi/PCr ratios. Dog hearts were more resistant than those of cats to changes in Pi/PCr with increasing work load. It is possible that animals adapted to different life-styles may have cardiovascular systems which are metabolically and mechanically adapted for different forms of stress. These differences may be elicited and effectively delineated using in vivo NMR techniques during various physiological interventions, such as pacing. The basis for these differences may be related to cardiac microvasculature or to intrinsic differences in enzyme kinetics. Delineation of these mechanisms may be helpful in the understanding of the physiological basis of cardiac function in health and disease.

Adaptation, Physiological↗

Is the ISA of pindolol beta 2-adrenoceptor selective?

1. Pindolol is a beta-adrenoceptor blocking drug with ISA (also called partial agonist activity). This means that in addition to blocking the effects of beta-adrenoceptor agonists, it produces some stimulation of beta-adrenoceptors. 2. In vitro studies with pindolol show that its maximum stimulant action is similar to that of isoprenaline in tissues possessing mainly beta 2-adrenoceptors, but is negligible in tissues possessing mainly beta 1-adrenoceptors. This suggests selective stimulation of beta 2-adrenoceptors. 3. In man the arteriodilator effects observed after intra-arterially infused pindolol at concentrations within the same range as those producing an antihypertensive effect also suggest a stimulant action on vascular beta 2-adrenoceptors. 4. The fact that pindolol prevents the reduction of resting heart rate and cardiac output observed after drugs lacking ISA at first sight suggests stimulation of cardiac beta 1-adrenoceptors. However, human atria possess not only beta 1- but also beta 2-adrenoceptors, stimulation of which would produce the same effect. 5. Although all beta-adrenoceptor antagonists lower blood pressure, recent experiments have shown that those agents with combined beta 1-adrenoceptor blocking activity and ISA at those receptors are less effective. This observation lends weight to the thesis that pindolol does not stimulate beta 1-adrenoceptors since it lowers blood pressure as effectively as drugs lacking ISA. 6. The evidence available therefore suggests that although pindolol blocks both beta 1- and beta 2-subtypes, it selectively stimulates beta 2-adrenoceptors.

Adrenergic beta-Antagonists↗

Is disordered folate metabolism the basis for the genetic predisposition to neural tube defects?

Vitamin levels were measured in twenty women under 35 years of age with a history of two or more neural tube defect pregnancies. Each index case was compared with a female control matched for age, obstetric history and social class. The mean concentration of red cell folate in the subjects was 178 ng/ml, significantly lower than the mean of 268 ng/ml for the control group (P = 0.005). Red cell folate levels showed a linear relationship with the number of neural tube defect pregnancies, the levels being lowest in women who had had three or four affected offspring. There was no significant difference in serum folate; plasma or white cell vitamin C; plasma vitamin A; thiamine, riboflavine or pyridoxine status; serum vitamin B12; plasma vitamin E; total protein, albumin, transferrin, magnesium, copper or zinc. Diet was assessed by a questionnaire. The dietary intakes of total folate and other vitamins except vitamin A were lower in the subjects than the controls but none of the differences were statistically significant. Regression analysis showed a difference between subjects and controls in the relationship of red cell folate to dietary folate. This study demonstrates an association between susceptibility to offspring with neural tube defects and depressed red cell folate levels which cannot be entirely attributed to a lower dietary intake of folate. It is postulated that one factor predisposing to the occurrence of neural tube defects may be an inherited disorder of folate metabolism.

Adult↗

Caries and treatment patterns in children related to school lunch program eligibility.

This article presents the caries prevalence and treatment patterns of children relative to a classification of family economic status, based on the child's eligibility for the free or reduced-cost school lunch program. Dental caries prevalence is reported for children aged 8-11 years at the beginning of a longitudinal clinical trial in the nonfluoridated city of Tampa, Florida. These children were placed into one of three school lunch subsidy categories--poverty, near-poverty, and nonpoverty--using federal government income eligibility guidelines for the school lunch program. The results support an inverse relation of caries prevalence to economic status, as well as the expected direct relation of family income to treatment needs met. The nonpoverty group exhibited the lowest number of decayed surfaces and had nearly 70 percent of their treatment needs met. The poverty group had significantly higher scores both for total caries experience (DMFS) and decayed surfaces (DS) than either of the other groups. Filled surfaces (FS) scores were not significantly different for any of the groups. This method of classifying children by school lunch program eligibility is considered valid because of the rigorous requirements used in determining family income, as well as the thorough verification procedure that is applied. The method is easy to use once officials have been convinced that confidentiality will be maintained.

Child↗

Is stimulation of prejunctional dopamine receptors an antihypertensive principle?

Bromocriptine and co-dergocrine (Hydergine R) reduce blood pressure and heart rate in experimental animals largely by stimulating DA2 receptors. A stimulant effect on DA1 receptors can be demonstrated in isolated tissues, but this action does not appear to make an important contribution to the depressor response to the two compounds, since effects are abolished by DA2 receptor blockade. Although both compounds are known to penetrate the brain, their antihypertensive effects, and the concomitant reductions in plasma noradrenaline levels produced in hypertensive patients can be prevented by domperidone, confirming a peripheral site of action. A review of the data available concerning the cardiovascular effects of bromocriptine and co-dergocrine in animals and man suggests that a DA2 receptor stimulant is effective in the treatment of hypertension.

Animals↗

Myocardial high energy phosphate metabolism in closed chest dog: creation of an animal model.

A chronic closed chest dog model was developed to study myocardial metabolism with NMR spectroscopy. Cardiac windows were surgically created in 10 dogs by removal of two ribs and accompanying skeletal muscle. Marlex mesh was sewn between the two exposed ribs, and fascia and skin were closed. 31P spectra were obtained using a surface coil placed into the surgically created pouch and using routine NMR pulsing techniques.

Adenosine Triphosphate↗

P NMR evaluation of hypoxic stress in brain of animal models.

Quantification by NMR of hypoxic stress is best afforded by the ratio of phosphocreatine (PCr) to inorganic phosphate (Pi). The thin cranium and sternum of neonates affords ideal conditions for NMR evaluation of impaired oxidative metabolism. Brains of animal models (dog and cat) are best studied with muscles retracted with the rf coil placed on the bare skull. 31P and 1H (lactate) signals are time shared with a doubly tuned coil. Servo-stabilized hypoxia gives simplified steady state analysis. Hypoxic insults of the brain are evaluated by the integral of the deviation of the PCr/Pi ratio from the normoxic values. Generally, a deviation of 1 unit of PCr/Pi for an interval of 3 hr will lead to a severe metabolic damage and recovery from hypoxia is often 30-fold delayed. Comprehensive monitoring of energy metabolism (PCr/Pi), respiratory chain redox states, intracellular pH and lactate accumulation allow the calculation of integrated pH and lactate "insults" and thus offer new insights on the effects of hypoxia on brain oxidative metabolism.

Animals↗

Multiple controls of oxidative metabolism in living tissues as studied by phosphorus magnetic resonance.

Three types of metabolic control of oxidative metabolism are observed in the various tissues that have been studied by phosphorous magnetic resonance spectroscopy. The principal control of oxidative metabolism in skeletal muscle is by ADP (or Pi/phosphocreatine). This conclusion is based upon studies of arm muscles of humans during steady-state exercise. A work-cost (Vm vs. Pi/phosphocreatine) relationship follows a Michaelis-Menten rectangular hyperbola, where Km values from 0.5 to 0.6 and Vmax values from 50 to 200 (at nearly constant pH) are found in linearized plots of the equation V/Vmax = 1/(1 + 0.6 phosphocreatine/Pi) where V is work level (which is equal to the velocity of the enzymatic reaction) and Vmax is the maximal work capacity that is a measure of the enzyme activity (E) of oxidative metabolism. Adaptation to exercise enhances the slope of the work-cost relationship and causes large changes in Vmax or E. A second metabolic control may enhance the slope of the work-cost relationship but not Vmax. For example, the initiation of exercise can lead to an improved characteristic that can be explained by 2-fold increased substrate delivery, for example, increased oxygen delivery by microcirculatory control. Cardiac tissue of the adult dog affords an example of optimal endurance performance adaptation and exhibits the steepest work-cost relationship observed and is attributed to a coordinated control of substrate delivery that may involve Ca2+ and inorganic phosphate control of NADH; control of O2 delivery may also be involved. The calculated work-cost relationship is similar to that observed in the beagle heart. The theoretical curve illustrates that the liability of multiple controls is a sharp break point in metabolic control at the end of the multiple control range--a possible cause of instability of cardiac performance at high V/Vmax.

Acidosis↗

Phosphorus magnetic resonance spectroscopy studies of the role of mitochondria in the disease process.

The incisive detection of bioenergetic insufficiency in an organ of known workload by P MRS is noninvasive and nondestructive, and in some cases the portion of the organ involved can be determined, particularly if both PCr and ATP are depleted. The fractional loss of ATP and hence the relative volumes of viable and "metabolically dead" tissue are thereby evaluated. In addition, the value of P MRS in following a therapy complements its value in diagnosis as this has been demonstrated in cases followed over 6 months to three years. The fact that deficiencies of the enzymes and substrates of oxidative metabolism can be detected by P MRS affords a global overview of energy metabolism that can be a key to rapid diagnosis. The distinction of the enzyme and/or substrate deficiency, while not directly indicated by steady state P MRS, can be identified by use of the "Crossover Theorem" and its impact upon blood and tissue levels of substrates (including oxygen). In the case of neonatal systemic hypoxia, there is no doubt about which of the equations applies, and similarly in metabolic disease, a glutaric acid urea is a direct consequence of the crossover response of metabolism and signifies that an enzyme deficiency may be involved. Furthermore, the clinical danger of a high Pi/PCr value is clarified by our observations, both from the animal models and from the theory, the high clues; i.e. 2 and over, suggest work stresses near the capability of oxidative metabolism and imminent failure of the negative feedback afforded by metabolic regulation, particularly ADP control of oxidative metabolism. This control is lost because of the fall of phosphocreatine to the point where creatine kinase is no longer in equilibrium, leading to the loss of ATP and its conversion to large amounts of ADP and its breakdown products. ATP then stimulates glycolysis and results in a massive lactic acidosis. At the same time, the low thermodynamic capability of glycolytic metabolism is unable to prevent irreversible ion disequilibration, water movements, edema, and eventually rupture of the cell membrane. The pathway of resynthesis of ATP is then tortuous, particularly as AMP is deaminated and adenosine is converted eventually to hypoxanthine. Thus, NMR reports that metabolic control is operating in the region where homeostasis of biochemical parameters is feasible. It further reports regions where the metabolic control is susceptible to failure and most aggressive clinical care is required.

Adenosine Triphosphate↗

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

Saccharomyces cerevisiae contains two genes, CIT1 and CIT2, encoding functional citrate synthase (K.-S. Kim, M. S. Rosenkrantz, and L. Guarente, Mol. Cell. Biol. 6:1936-1942, 1986). We show here that CIT2 encodes a nonmitochondrial form of citrate synthase. The DNA sequence of CIT2 presented provides a possible explanation for why the CIT2 product, unlike the CIT1 product, fails to be imported into mitochondria. While the products of these two genes are highly homologous, they diverge strikingly at their amino termini. The amino terminus of the CIT1 primary translation product extends 39 residues beyond the amino termini of Escherichia coli and porcine citrate synthases. This extension consists of a typical mitochondrial targeting motif. The amino terminus of the CIT2 primary translation product extends 20 residues beyond the amino termini of the E. coli and porcine enzymes. The CIT2-encoded extension is not homologous to that of CIT1, resulting in a nonmitochondrial localization of the product. The CIT2-encoded extension, however, does bear certain similarities to mitochondrial targeting sequences. The possible role of this sequence in targeting this CIT2 product to a nonmitochondrial organelle is discussed.

Amino Acid Sequence↗

Diagnosis and therapeutic evaluation of a pediatric case of cardiomyopathy using phosphorus-31 nuclear magnetic resonance spectroscopy.

An 8 month old girl presented with undiagnosed non-anatomic congenital cardiomyopathy with massive cardiomegaly on chest X-ray film. Her older sibling had died suddenly at 6 months of age from what appeared to be a similar abnormality. Utilizing phosphorus-31 nuclear magnetic resonance (P-31 NMR) surface coil spectroscopy, a metabolic disorder was demonstrated in both her myocardium and skeletal muscle, revealing a phosphocreatine (PCr) to inorganic phosphate (Pi) ratio of half of that for a normal control infant. Manipulation of serum substrate availability indicated that medium chain triglycerides alone did not improve myocardial metabolism, but that intravenous glucose or oral carbohydrate loading raised the myocardial PCr/Pi ratio from 1.0 +/- 0.05 to 1.8 +/- 0.1 (p less than 0.01) without significantly affecting the PCr/Pi value of her resting skeletal muscle. This study demonstrates the feasibility of using P-31 nuclear magnetic resonance to evaluate the biochemistry of the human myocardium in vivo and to diagnose a metabolic abnormality. Phosphorus-31 nuclear magnetic resonance can thus be used to optimize therapy for human disease.

Cardiomegaly↗

Control of oxidative metabolism and oxygen delivery in human skeletal muscle: a steady-state analysis of the work/energy cost transfer function.

The concept of transfer function for organ performance (work output vs. biochemical input) is developed for skeletal and cardiac muscle under steady-state exercise conditions. For metabolic control by the ADP concentration, the transfer function approximates a Michaelis-Menten hyperbola. Variation of the work identifies metabolic operating points on the transfer function corresponding to ADP concentrations or to a ratio of inorganic phosphate to phosphocreatine that can be determined by phosphorus nuclear magnetic resonance. This operating point is characterized by the fraction (V/Vmax) of maximal activity of oxidative metabolism in the steady state. This quantity appears to be useful in predicting the degree to which metabolic homeostasis is effective; poorly controlled metabolic states can readily be identified and are used in the diagnosis and therapy of metabolic disease in the organs of neonates and adults.

Adenosine Diphosphate↗