Histamine H3 receptor-mediated inhibition of sympathetically evoked mydriasis in rats.

This study was designed to determine if the histamine H3 receptor agonist R-alpha-methylhistamine would play a role in modulation of sympathetically evoked mydriasis in anesthetized rats, and if so, to ascertain the specific receptor subtype(s) involved. Reproducible frequency-response curves of pupillary dilation were generated by stimulation of the cervical preganglionic sympathetic nerve (1-32 Hz). Systemic administration of R-alpha-methylhistamine (0.3-3.0 mg kg(-1)) produced a dose-related inhibition of the evoked mydriasis. The greatest inhibition was seen at lower frequency levels, with about 43% depression observed at 2 Hz. The specific histamine H3 receptor antagonist, clobenpropit (3.0 mg kg(-1), i.v.), blocked the inhibitory effect of R-alpha-methylhistamine, whereas neither the histamine H2 receptor antagonist, cimetidine (5.0 mg kg(-1), i.v.), nor the histamine H1 receptor antagonist, chlorpheniramine (0.5 mg kg(-1), i.v.), was effective. The histamine H2 receptor agonist, dimaprit (10 mg kg(-1), i.v.), was also without effect on the evoked mydriasis. R-alpha-methylhistamine (3.0 mg kg(-1)) did not inhibit phenylephrine-induced mydriasis. These results support the conclusion that R-alpha-methylhistamine produces inhibition of sympathetically evoked mydriasis via histamine H3 receptor stimulation, presumably by an action on presynaptic histamine H3 receptors.

Sympathetic control of nasal blood flow in the rat mediated by alpha(1)-adrenoceptors.

Experiments were undertaken, using laser-Doppler flowmetry, to determine the nature of adrenoceptors mediating sympathetic nerve evoked nasal vasoconstrictor responses in anesthetized rats. Presence of sympathetic tone was confirmed by a large (330%) increase of nasal blood flow following section of the ipsilateral preganglionic cervical sympathetic nerve. Electrical nerve stimulation produced reproducible, frequency-related nasal vasoconstrictor responses with near maximal response, observed at less than 10 Hz. Evoked nasal vasoconstrictor responses were largely blocked with intravenous treatment with the non-selective alpha-adrenoceptor antagonists, phentolamine (5 mg kg(-1)) and phenoxybenzamine (2 mg kg(-1)), as well as with the selective alpha(1)-adrenoceptor antagonist, prazosin (300 microg kg(-1)). alpha(2)-Adrenoceptor antagonism with rauwolscine (500 microg kg(-1)) potentiated neurally evoked nasal vasoconstriction. Neither atropine (1 mg kg(-1)) nor propranolol (1 mg kg(-1)) altered the evoked responses. Rats with intact cervical sympathetic nerves responded to rauwolscine with a modest constriction. Subsequent prazosin administration produced an increase of nasal blood flow of approximately 275%. These results suggest that the nasal vasculature of the rat is under intense sympathetic tone and that the resulting neurogenic vasoconstriction is mediated exclusively by activation of alpha(1)-adrenoceptors.

Inhibition of human telomerase by nucleotide analogues bearing a hydrophobic group.

Telomerase, which synthesizes telomeric DNA in eukaryotic cells, is classified as a reverse transcriptase. To clarify the susceptibility of telomerase to nucleoside 5'-triphosphates bearing a hydrophobic group on the base moiety, we studied the inhibitory effects of 2',3'-dideoxy-5-styryluridine 5'-triphosphate analogues and 9-(beta-D-arabinofuranosyl)-2-(p-n-butylanilino)purine 5'-triphosphate analogues on telomerase activity using a quantitative 'stretch PCR' assay. 2',3'-Dideoxy-5-styryluridine 5'-triphosphate (StddUTP) showed more potent inhibition than 2',3'-dideoxythymidine 5'-triphosphate (ddTTP). On the other hand, 9-(beta-D-arabinofuranosyl)-2-(p-n-butylphenyl)guanine 5'-triphosphate (BuParaGTP) showed no inhibition, even though 9-(beta-D-arabinofuranosyl)guanine 5'-triphosphate (araGTP) is a potent inhibitor of telomerase. The influence on telomerase of hydrophobic substituents on the base moieties of nucleotides is described.

Sympathetic vasodilation in the rat anterior choroid mediated by beta(1)-adrenoceptors.

Electrical stimulation of the preganglionic superior cervical nerve produced a frequency-dependent vasoconstrictor response in the anterior choroidal blood vessels of the eye of anesthetized rats. Systemic administration of phentolamine (5 mg kg(-1)) reversed the vasoconstriction to a vasodilator response. This sympathetic-evoked vasodilation was not antagonized by inhibition of nitric oxide synthase with N(G)-nitro-L-arginine methyl ester (L-NAME) (20 mg kg(-1)) or by inhibition of cyclo-oxygenase with indomethacin (20 mg kg(-1)). Intravenous administration of propranolol (1 mg kg(-1)), as well as selective beta(1)-adrenoceptor antagonists atenolol (3 mg kg(-1)), timolol (0.3 mg kg(-1)), and betaxolol (0.1 mg kg(-1)), totally abolished the sympathetic nerve evoked ocular vasodilation. In contrast, the selective beta(2)-adrenoceptor antagonist, ICI-118, 551 ((+/-)-1-[2, 3-(Dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl)amino]-2- butanol) (0.3 mg kg(-1), i.v.), was without effect. These results support the conclusion that the residual sympathetic ocular vasodilation observed in the rat anterior choroid after alpha-adrenoceptor blockade is mediated exclusively by neurogenic release of norepinephrine acting on vascular beta(1)-adrenoceptors.

Sympathetic vasoconstriction in the rat anterior choroid is mediated by alpha1-adrenoceptors.

These experiments were undertaken in an attempt to use laser-Doppler flowmetry to measure anterior choroidal blood flow in the anesthetized rat and to study the mechanism by which sympathetic nerve stimulation might produce vasoconstriction in this vascular bed. Electrical stimulation of the preganglionic cervical sympathetic nerve produced reproducible, frequency-related ocular vasoconstrictor responses with maximal vasoconstriction seen at about 32 Hz. Ocular vasoconstrictor responses were blocked by intravenous treatment with the nonselective alpha-adrenoceptor antagonists, phentolamine (5 mg kg(-1)) and phenoxybenzamine (2 mg kg(-1)), as well as with the selective alpha1-adrenoceptor antagonist, prazosin (0.3 mg kg(-1)). In contrast, the selective alpha2-adrenoceptor blocker, rauwolscine (0.5 mg kg(-1)), only potentiated the vasoconstriction. Neither intravenous atropine (1 mg kg(-1)) nor propranolol (1 mg kg(-1)) altered the magnitude of neurally evoked vasoconstriction. These results demonstrate the usefulness of laser-Doppler flowmetry in studies of the rat anterior choroidal circulation and suggest that adrenergic neurogenic vasoconstriction in the anterior segment of the rat eye is mediated almost exclusively by alpha1-adrenoceptor mechanisms.

Use of microbial electrodes for observation of microbiological nitrogen elimination process.

Microbial electrodes for the determination of ammonia and the estimation of biochemical oxygen demand (BOD) were applied to the nitrogen elimination process. The dimensions of the nitrification and the denitrification vessels were 170 and 70 L, respectively. The wastewater used for the experiment was obtained from a fermentation factory and adjusted to 470-530 mg/L of total Kjeldahl nitrogen and 1700-3000 mg/L of BOD. The ammonia electrode was assembled with a membrane containing nitrifying bacteria and an oxygen probe. The BOD electrode was similarly constructed, except it used a membrane containing the yeast, Trichosporon cutaneum. A flow system was employed for the automatic measurement of samples every 30 min. The nitrification and denitrification rates of the activated sludge were measured to investigate the optimum conditions and evaluate the capacity of the plant. The various data obtained by the microbial electrodes allowed us to inspect the situation of the plant and estimate control parameters such as nitrogen and BOD loadings. The average removals of ammonia nitrogen and total Kjeldahl nitrogen were 96% and 89%, respectively, during the experiment for period of 2 weeks.

Neurogenic cutaneous vasodilation in the cat forepaw.

The present experiments were undertaken to determine, using Laser Doppler flowmetry, if elimination of efferent constrictor mechanisms would unmask cutaneous vasodilator responses following preganglionic sympathetic nerve stimulation in the forepaw of anesthetized cats. We also addressed the question of a potential causal relationship between neurally evoked vasodilator and sudomotor responses. Three separate anti-adrenergic regimens were utilized: (1) acute guanethidine administration (1-2 mg/kg); (2) chronic monoamine depletion with reserpine (5 mg/kg) and alpha-methyl-para-tyrosine (2 x 300 mg/kg); and (3) alpha-adrenoceptor blockade with prazosin (300 micrograms/kg) and yohimbine (0.5 mg/kg). Guanethidine treatment produced a significant depression of basal cutaneous blood flow whereas alpha-adrenoceptor blockade did not. In all three groups, stimulation of the preganglionic thoracic sympathetic nerve trunk produced intensity-dependent increases of digital skin blood flow along with near-maximal sympathetic-cholinergic sudomotor (electrodermal) responses recorded simultaneously from the same paw. Vasodilator responses were not altered by intravenous propranolol (1 mg/kg) or atropine (1 mg/kg); however, evoked sudomotor responses were totally blocked by atropine. Low doses (1.5 mg/kg i.v.) of hexamethonium selectively abolished the cutaneous vasodilator responses but not concomitantly evoked sudomotor responses. These results demonstrate, using direct measurements of blood flow, that cutaneous digital vasodilation can be measured in cats following removal of vasoconstrictor mechanisms either pre- or postjunctionally. Neither muscarinic nor beta-adrenoceptor mechanisms appear to be involved. These experiments also suggest that cutaneous vasodilation is not a consequence of concomitant sudomotor activation.

Neural activation of alpha-2 adrenoceptors in cat cutaneous vasculature.

The present study was designed to assess the relative contribution of postjunctional alpha-1 and alpha-2 adrenoceptors in neurally evoked cutaneous vasoconstrictor responses in anesthetized cats. Preganglionic stimulation of the thoracic sympathetic nerve trunk produced an intensity-related decrease of digital skin blood flow as measured by laser-Doppler flowmetry. Sympathetic-cholinergic sudomotor (electrodermal) responses were recorded simultaneously as an additional index of neuronal activation. Vasoconstrictor responses were not altered by pretreatment with i.v. propranolol (1 mg/kg) or atropine (1 mg/kg) and were refractory to low doses (2 mg/kg) of hexamethonium. As expected, atropine abolished evoked skin potential responses which were also sensitive only to higher doses (20 mg/kg) of hexamethonium. Pretreatment with either phentolamine (1 mg/kg i.v.), yohimbine (0.5 mg/kg i.v.) or prazosin (0.3 mg/kg i.v.) produced significant reduction of sympathetic vasoconstriction with blockade by phentolamine and yohimbine being far greater than that seen with prazosin. In animals previously given prazosin (0.3 mg/kg), subsequent administration of yohimbine (0.5 mg/kg) almost totally blocked the remaining evoked cutaneous vasoconstriction. These results demonstrate that both alpha-1 and alpha-2 adrenoceptors are present in the cutaneous bed of the cat with the predominant effect mediated by innervated alpha-2 adrenoceptors.

Suppression of clonidine-induced vasoconstriction by cooling.

Using the cannula inserting method, we investigated whether vascular responses to norepinephrine, phenylephrine, clonidine, tyramine and KCl were altered by cooling (37 degrees C to 27 degrees C) in isolated canine ear arteries. Vasoconstrictor responses to norepinephrine, phenylephrine and tyramine were slightly depressed or unchanged, whereas those to clonidine and KCl were significantly suppressed by cooling. It is suggested that activation of Ca channels via alpha-2 adrenoceptors may be depressed by cooling in dog ear arteries.