Biomedical subjects
Publications and source records attributed to D A Taylor.
The primary sequence of a cDNA encoding a novel transglutaminase from a human prostate cDNA library is reported. The sequence is compared to other known transglutaminases in a multiple alignment. The deduced peptide sequence is 51% identical to a rat prostate transglutaminase.
Rats were exposed to increasing concentrations of morphine hydrochloride (up to 0.4 mg/ml) in 5% w/v sucrose solution as their sole source of drinking water. Physical dependence was established as determined by the precipitation of withdrawal behaviour following administration of 1 mg/kg IP naloxone hydrochloride on day 23. The choice between either a 5% w/v sucrose solution or a 5% w/v sucrose solution containing 0.4 mg/ml morphine hydrochloride 4 days following withdrawal resulted in rats being categorized into two groups based on their respective consumption of the morphine-containing solution. The amount of morphine solution voluntarily consumed by approximately half the rats were sufficiently high as to lead to a relapse into physical dependence to morphine. The high preference for morphine shown by these rats could not be attributed to the taste of the morphine solution. Naive rats or rats exposed to a 5% w/v sucrose solution for 23 days failed to consume significant quantities of the morphine-containing solution when provided with a choice. The administration of either an IM slow-release formulation of 70.5 mg/kg haloperidol decanoate (= 50 mg/kg haloperidol) or 10 micrograms/kg IP ondansetron hydrochloride daily did not alter morphine ingestion in the high morphine-preferring rats.
Using an in situ isolated salt-perfused rabbit lung preparation, we investigated the functional ontogeny of pulmonary vascular dopamine receptors. In rabbits from 1 to 23 d of age, we measured pulmonary vascular vasodilatory responses to the peripheral vascular dopamine receptor (DA1) agonist, fenoldopam, and sodium nitroprusside during prostaglandin F2 alpha-induced pulmonary vasoconstriction. In separate experiments, the lungs were pretreated with the DA1 receptor blocker, SCH 23390, before prostaglandin F2 alpha, fenoldopam, and sodium nitroprusside. Lungs from rabbits at one of 6 age groups (n = 6-8 per group) were ventilated and perfused. After a stabilization period, prostaglandin F2 alpha was infused into the pulmonary inflow catheter in a concentration range to yield a sustained rise in mean pulmonary artery pressure (4.9 +/- 0.2 mm Hg). Fenoldopam was injected into the pulmonary artery at doses of 0.01, 0.1, 1.0, and 10 micrograms/g after a recovery period, sodium nitroprusside (0.2 micrograms/g) was injected into the pulmonary artery, and the resultant changes in vascular pressure were recorded. Across all age groups, with and without DA1 receptor blockade, sodium nitroprusside-induced vasodilation was similar (-2.7 +/- 0.2 mm Hg) and was considered reference vasodilation. The fenoldopam vasodilation response was considered a percentage of the sodium nitroprusside reference. Response to fenoldopam varied significantly (p < 0.05 by analysis of variance) across the six age groups, with a maximum at 3-5 d of age. Pretreatment with SCH 23390, a selective DA1-blocking agent, significantly attenuated fenoldopam vasodilation in all but the youngest animals (age 0-2 d), in which no blockade effect was noted.(ABSTRACT TRUNCATED AT 250 WORDS)
In the present study, we compared the blood pressure in the SHR-SP and in the spontaneously hypertensive rat (SHR) after dietary administration of fish oil from 4 to 17 weeks of age. The retarding influence of dietary fish oils on the development of hypertension was prominent in the SHR (26 mmHg) and not evident in the SHR-SP (8 mmHg). The enhanced development of blood pressure in both the SHR and the SHR-SP is characterised by an elevated maximum contraction in the mesenteric vascular bed to sympathetic nerve stimulation and to injected noradrenaline. In SHR, but not SHR-SP, this maximum contraction was significantly attenuated by dietary fish oil. Likewise, acetylcholine mediated relaxation of the isolated aorta was enhanced in preparations from the SHR but not the SHR-SP. These physiological changes were also associated with a change in the total n-3 polyunsaturated fatty acids (PUFAs) content in vascular tissue, which were inversely proportional to the prevailing blood pressure values seen in all three strains of rat receiving dietary fish oils. Platelet activated thromboxane production was equally depressed in WKY (Wistar Kyoto), SHR and SHR-SP rats. The results indicate that the blood pressure lowering effect of fish oil when administered during the period of development of hypertension is much greater in the SHR than it is in the SHR-SP. Furthermore the lowering of blood pressure by fish oil administration is related to a restoration of normal vascular contraction and normal vascular relaxation, but not related to a suppression of serum thromboxane production.
Because alpha-1 adrenoceptor subtypes are distributed differentially in different arteries, experiments were conducted to determine the functional contribution of these subtypes in conduit vs. resistance vessels. Concentration- or dose-response curves for norepinephrine were obtained from aortic rings, superior mesenteric artery rings or the isolated perfused mesenteric vasculature from male Sprague-Dawley rats. Frequency-response curves to transmural electrical stimulation were obtained from the latter two preparations. The effects of 5-methylurapidil (5-MU), an alpha-1a adrenoceptor antagonist, and chloroethyl-clonidine (CEC), an alpha-1b adrenoceptor antagonist, on contractile responses were determined. In artery rings, 5-MU (30 nM) had no effect on the EC50 of norepinephrine but reduced the maximum response of the mesenteric artery rings by nearly 25%. In the perfused mesenteric vasculature, however, 5-MU (30 nM) shifted the ED50 for norepinephrine about 40-fold while reducing the maximum response by 30%. 5-MU depressed the frequency-response curve in the perfused mesenteric vasculature by nearly 80%, but did not alter the response of artery rings. In aorta, pretreatment with CEC (10 microM) shifted the concentration-response curve of norepinephrine by 800-fold without effecting the maximum. In mesenteric artery rings and perfused mesenteric vasculature, CEC reduced the slope and maximum response of both frequency-response and norepinephrine dose-response curves. Responses to norepinephrine (10 microM) in the perfused mesentery were abolished by 5-MU and reduced only 25% by nifedipine. These data suggest that the density or role of alpha-1a adrenoceptors may be greater in resistance vessels than in conduit vessels.
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A highly efficient and reproducible two-step liquid-liquid ion-pair extraction technique for the isolation of morphine from biological samples is described. A rapid normal phase high-performance liquid chromatographic procedure coupled with amperometric electrochemical detection has also been developed for subsequent quantification of morphine. Extraction involves the disruption of brain tissue or plasma in methanol, centrifugation, evaporation and reconstitution in ethyl acetate containing 10 mM di-(2-ethylhexyl) phosphoric acid, a liquid cation-exchanger, and back-extraction into 170 mM orthophosphoric acid. An acidic eluent consisting of acetonitrile-76 mM orthophosphoric acid-ammonia buffer (pH 3.0) (15:85, v/v) in combination with a strong cation-exchange column allows complete separation of morphine and the internal standard, nalbuphine. The limit of detection for morphine is 1.3 ng on-column.
The role of axon-target cell interactions in shaping the prevalence and distribution of synaptic densities was studied in an experimentally-induced aberrant functional projection from the chick cochlear nucleus (nuc. magnocellularis, NM) to the contralateral NM. Contact with an abnormal target appears to induce in the aberrant axons a pattern of presynaptic densities resembling that in normal cochlear nerve endings in NM. NM axon terminals induced similar numbers of postsynaptic densities (PSDs) per unit length of membrane apposition in both their normal and abnormal targets but the longer membrane apposition in the highly invaginated aberrant terminal in NM results in a significantly greater amount of postsynaptic density per ending. These auditory neurons thus appear able to adjust a variety of features to permit assembly and maintenance of a novel functional synapse.
Three primary pigmented pheochromocytomas of the adrenal gland are presented. The pigment in all cases proved to be melanin. Two of the pheochromocytomas were sporadic and histologically typical, except for a focal spindle cell configuration in one. It is believed that the morphologic appearance of these tumors represents divergent differentiation from neural crest, expressing typical pheochromocytoma (polygonal cells) and melanocytic features (melanin pigment).
The observation that adenovirus E1A gene products can inhibit differentiation of skeletal myocytes suggested that E1A may interfere with the activity of myogenic basic helix-loop-helix (bHLH) transcription factors. We have examined the ability of E1A to mediate repression of the muscle-specific creatine kinase (MCK) gene. Both the E1A12S and E1A13S products repressed MCK transcription in a concentration-dependent fashion. In contrast, amino-terminal deletion mutants (d2-36 and d15-35) of E1A12S were defective for repression. E1A12S also repressed expression of a promoter containing a multimer of the MCK high-affinity E box (the consensus site for myogenic bHLH protein binding) that was dependent, in C3H10T1/2 cells, on coexpression of a myogenin bHLH-VP16 fusion protein. A series of coprecipitation experiments with glutathione S-transferase fusion and in vitro-translated proteins demonstrated that E1A12S, but not amino-terminal E1A deletion mutants, could bind to full-length myogenin and E12 and to deletion mutants of myogenin and E12 that spare the bHLH domains. Thus, the bHLH domains of myogenin and E12, and the high-affinity E box, are targets for E1A-mediated repression of the MCK enhancer, and domains of E1A required for repression of muscle-specific gene transcription also mediate binding to bHLH proteins. We conclude that E1A mediates repression of muscle-specific gene transcription through its amino-terminal domain and propose that this may involve a direct physical interaction between E1A and the bHLH region of myogenic determination proteins.
A 28-year-old white male presented with a palpable abdominal mass and a grossly elevated beta human chorionic gonadotrophin. He refused investigations to establish the pathology and was treated with cisplatin combination chemotherapy. After an initial fall in his marker, it rose while he was on chemotherapy. The tumour was resected which confirmed the presence of a retroperitoneal germ cell tumour (extra-gonadal). He received further adjuvant chemotherapy and is subsequently clear of disease after being off treatment for more than 5 years. Total resection is not usually considered possible in patients with persistent positive markers after induction chemotherapy. This case shows that surgery should be considered as the results of salvage chemotherapy are poor in these patients.
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Adaptive supersensitivity has been demonstrated previously in the guinea pig vas deferens after chronic treatment with reserpine, postganglionic denervation, or preganglionic denervation. The magnitude of the change in sensitivity was similar regardless of the method of induction; the underlying mechanism was identified as a partial depolarization secondary to reduced activity of the Na+/K+ pump. Experiments were conducted to quantitatively determine whether the identified losses in Na+/K(+)-ATPase activity and [3H]ouabain binding were due to reductions in the levels of specific protein subunits of the sodium pump. Electrophoretic separation and quantification of the abundance of alpha subunit isoforms were accomplished using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and slot blot analysis. Supersensitivity was induced in the guinea pig vas deferens through pretreatment with reserpine (1.0 mg/kg/day x 5 days). The abundance of the alpha 2 subunit isoform was reduced by 41% in tissue homogenates obtained from animals treated with reserpine, compared with untreated controls. In contrast, there was no significant alteration in the alpha 1 subunit isoform (a protein similar in size to that previously identified in vascular smooth muscle as a "truncated" form of the protein). These data suggest that the adaptation of the guinea pig vas deferens after a chronic reduction in net stimulus is mediated through a change in a specific cellular protein. This evidence supports the assignment of the alpha 2 subunit isoform as the specific protein responsible for the development of nonspecific adaptive supersensitivity in the guinea pig vas deferens.
beta-Endorphin and morphine produce an increase in the latency of the tail-flick reflex when administered into the PAG of awake rats. The antinociceptive effect of both opioid agonists was blocked by the sequential local injection of either CTP (D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-NH2), a selective mu opioid receptor antagonist, naltrexone, or beta-endorphin (1-27), a putative epsilon opioid receptor antagonist, with minimal selectivity. When either CTP or naltrexone was used as the antagonist, the dose-inhibition curves generated for beta-endorphin and morphine were not parallel, suggesting the involvement of separate and distinct receptors. Also, synergism occurred when a dose of morphine producing submaximum antinociception was administered simultaneously with either a submaximal or ineffective dose of beta-endorphin. Inhibition of the antinociceptive response to beta-endorphin by mu antagonists and the non-selective antagonism of both beta-endorphin and morphine by beta-endorphin (1-27) suggested that epsilon opioid receptors were not involved. Additionally, a mu/delta opioid receptor complex was not involved, since ICI 174,864 (Allyl2-Tyr-Aib-Aib-Phe-Leu-OH), a selective delta opioid receptor antagonist, did not alter the response to beta-endorphin. Thus, although additional characterization is required, beta-endorphin and morphine appear to act (at least in part) through different opioid receptors, demonstrable using selected mu opioid receptor antagonists.
Using an in situ isolated salt-perfused rat lung preparation, we investigated the pulmonary vascular response to atrial natriuretic peptide (ANP). ANP was infused in graded doses ranging from 0.01 to 100.0 micrograms/kg during prostaglandin F2 alpha-induced pulmonary vasoconstriction. These experiments were repeated after selective dopamine1 (DA1) receptor blockade with SCH 23390 and after catecholamine depletion by reserpine. ANP at doses of 0.01, 0.1, 1.0, 10.0, or 100.0 micrograms/kg was injected into the pulmonary artery (n = 5-7/dose). In the unblocked group ANP infusion resulted in a dose-dependent decrease in the mean pulmonary arterial pressure (PAP) with a maximum effect at 10 micrograms/kg (delta PAP = -3.4 +/- 0.2 mm Hg). In the DA1 blockade groups the ANP dose-response curve was shifted to the right, in a parallel fashion. After catecholamine depletion with reserpine, the ANP dose-response curve was identical to that of the unblocked groups. With the parallel, rightward shift of the ANP dose-response curves by SCH 23390 and no attenuation of ANP effect after catecholamine depletion, it appears that ANP vasoactive properties in the pulmonary vasculature involve an interaction with the vascular DA1 receptors. This observation differs from the ANP-dopaminergic interactions seen in the kidney in which ANP action depends on endogenous dopamine transmission.
Chronic treatment with reserpine (0.1 mg/kg/day x 7 days) leads to the development of adaptive supersensitivity of ventricular myocardium of guinea pigs. The compensatory increase in sensitivity is associated with a small increase in beta-adrenoreceptor number. However, sensitivity is increased to a number of agonists that do not interact with beta-adrenoceptors. An evaluation of the role of both adenylyl cyclase and guanine nucleotide-binding regulatory proteins in the development of adaptive supersensitivity was carried out using crude membrane fragments from untreated control and chronically reserpine-treated guinea pigs. Quantitative analysis of Gs alpha and Gi protein concentrations was accomplished using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Chronic treatment with reserpine reduced basal levels of adenylyl cyclase activity by nearly 60%. The reduced activity was not the result of a loss of endogenous norepinephrine, because incubation of tissues in the presence of propranolol did not alter the basal level of adenylyl cyclase activity. Incubation in the presence of guanylylimido diphosphate (10(-5) M) also significantly reduced basal adenylyl cyclase activity, by nearly 70%. Chronic treatment with reserpine failed to significantly alter the activation of adenylyl cyclase by isoproterenol, impromidine, NaF, or forskolin. These data suggest that chronic treatment with reserpine does not alter agonist-induced activation of adenylyl cyclase. Furthermore, analysis of Gs alpha and Gi indicated that chronic treatment with reserpine did not affect the levels of these regulatory proteins in ventricular myocardial membranes. The data indicate that the enhanced sensitivity of guinea pig ventricular myocardium is not the result of an alteration in adenylyl cyclase activity or in the concentration of guanine nucleotide regulatory proteins. Therefore, the enhanced responsiveness to widely diverse agonists must be due to an alteration in cellular function beyond the level of adenylyl cyclase.
Low-dose, chronic administration of the synthetic glucocorticoid, dexamethasone, to male CD-I rats in the drinking water or by osmotic minipumps implanted subcutaneously after supralethal irradiation of the kidneys (20 Gy) was studied. The effectiveness of treatment with dexamethasone in the drinking water at concentrations of 23 micrograms/l to 188 micrograms/l and treatment times varying from 33 to 166 days was evaluated. At monthly intervals kidney function was assayed by measuring the clearance of 51Cr-ethylenediaminetetraacetic acid. All dexamethasone treatment regimens increased survival times significantly and delayed the development of kidney dysfunction. The most effective combination of concentration of dexamethasone in drinking water and treatment interval after irradiation with respect to survival was 94 micrograms/l and 88 days. However, a slightly longer survival and a better functional result was obtained if an equivalent amount of dexamethasone was administered by minipumps. Shielding the adrenal glands during kidney irradiation did not prolong survival. This shows that the beneficial effect of dexamethasone is not due to compensation for reduced adrenal glucocorticoid production resulting from concomitant exposures of these glands during kidney irradiation.
Chronic treatment of guinea pigs with morphine produces subsensitivity (tolerance) of the longitudinal smooth muscle-myenteric plexus preparation to a variety of inhibitory agonists (e.g., mu opioid, alpha adrenoceptor and adenosine receptor agonists) and supersensitivity (dependence) to a variety of excitatory agonists (e.g., nicotine, 5-hydroxytryptamine and potassium ions). The present investigation was to determine if these changes in sensitivity could be related to changes in electrical properties of the S and AH neurons in the myenteric plexus. S neurons from morphine-implanted animals were significantly depolarized (7 mV) relative to those from placebo-implanted animals, whereas the membrane potential of AH neurons was unchanged. Approximately 60% of S neurons were hyperpolarized by morphine. In this subset of neurons, membranes were significantly depolarized but the threshold was unchanged in morphine-implanted animals. This means that resting potentials of S neurons from tolerant preparations are closer to threshold. The hyperpolarization produced by morphine (0.1 microM) was similar in preparations from morphine- and placebo-implanted animals. Thus, the partially depolarized state of S neurons in the myenteric plexus is the cause of the subsensitivity and supersensitivity to agonists and can explain both tolerance and dependence. Changes in opioid receptors or their coupling to potassium channels do not appear to contribute to tolerance in the longitudinal smooth muscle-myenteric plexus.