bluuebunny/crossref_metadata_2025 huggingface dataset:
α-Toxin perfusion: a new method for selective impairment of endothelial function in isolated vessels or intact vascular beds
Authors: Laher I. , Thorin-Trescases N. , Ding A. , Laporte R. , Osol G.
Published: April 2011

We describe a method for selectively permeabilizing endothelial ceils, using the membrane pore forming exoprotein Staphylococcus aureus α-toxin. Experiments were performed in rabbit central ear artery or its main side branch under isometric conditions, on the isolated perfused kidney, or in cannulated pressurized renal arteries. In presence of α-toxin, endothelial-dependent vasodilator responses elicited by acetylcholine or A23187 were abolished, whereas the sensitivity of smooth muscle cells to constrictors (norepinephrine, phenylephrine, or KCl) or dilators (sodium nitroprusside) was not affected. The results indicate that restricting the α-toxin to the luminal surface induces selective impairment of vascular endothelial function. This method of eliminating endothelium-dependent vasodilator responses may prove to be useful in the study of endothelial – smooth muscle interactions of isolated small arteries and intact vascular beds.Key words: α-toxin, endothelium, vasodilation, vascular smooth muscle.
Interferon-mediated changes in the expression of CYP1A1 in human B Iymphoblastoid (AHH-1 TK +I−) cells
Authors: Delaporte Enock , Renton Kenneth W. , Cribb Alastair E.
Published: April 2011

The expression of constitutive and inducible cytochrome P450s has been shown to be downregulated by interferon through an unknown pretranslational mechanism that depresses the mRNA encoding P450 apoproteins. To establish an association between gene transcription and P450 apoprotein downregulation by interferon, we studied the effect of recombinant interferon (IFN-α2α) on CYP1A1 in human B lymphoblastoid cell lines. The cHoI cell line expresses inducible native CYP1A1, while the genetically engineered derivative h1A1 v2 expresses a noninducible extrachromosomal vector-derived human CYP1A1 cDNA lacking the CYP1A1 promoter region. We characterized CYP1A1 activity, apoprotein, and mRNA by ethoxyresorafin O-deethylase activity, Western immunoblotting, and Northern blot analysis, respectively. In cHoI cells, following induction with dibenz[a,h]anthracene, interferon depressed CYP1A1 apoprotein and mRNA levels by 55 and 76%, respectively, with no detectable changes in enzyme activity. In h1A1 v2, however, interferon increased CYP1A1 activity, apoprotein, and mRNA. The depression of CYP1A1 mRNA and apoprotein levels incHoI cells, in contrast with the increase observed in h1A1 v2 cells, suggests that nuclear mechanisms are essential for interferon-mediated depression of inducible P450s. From our preliminary results we propose that interferon-mediated downregulation of CYP1A1 may result from inhibition of gene transcription.Key words: cytochrome P450, lymphoblastoid cells, interferon, transcription.
Characterization of nitroglycerine-induced relaxation in human corpus cavernosum smooth muscle: implications to erectile physiology and dysfunction
Authors: Christ George J. , Kim Daniel C. , Taub Harvey C. , Gondré C. Marjorie , Melman Arnold
Published: April 2011

The importance of the nitric oxide – guanylate cyclase – cGMP system in modulating corporal smooth muscle tone and penile erection has been amply demonstrated. The goal of these studies was to evaluate the possibility that age- or disease-related alterations in human corporal smooth muscle responsivity to activation of this pathway might play a role in the etiology of erectile dysfunction. Thus, we utilized a previously described heuristic model to assess the kinetic and steady-state characteristics of relaxation of precontracted isolated corporal tissue strips elicited by nitroglycerine (NTG). Studies were conducted on corporal tissue strips excised from 26 patients with organic erectile dysfunction, and 7 patients with documented erections. For the purposes of statistical analysis the impotent patient population was stratified into two age groups (A, ≤59 years; B, ≥60 years) and further subdivided into two diagnostic categories, diabetic and nondiabetic patients, respectively. In ≈75% of precontracted corporal tissue strips derived from impotent patients (contracted to ≈75% of maximum with phenylephrine), the NTG-induced response was biphasic, consisting of a rapid relaxation response that reached steady state before onset of a more slowly developing regaining of tension, termed the desensitization response. In contrast, a biphasic response was observed much less frequently (≈30%) in corporal tissue strips derived from a potent patient population (p < 0.0001). Statistical analysis revealed significant heterogeneity among corporal tissue strips derived from patients with organic erectile dysfunction, with respect to both the kinetic and steady-state characteristics of the NTG-induced relaxation and desensitization responses. In particular, the maximal rate constant for both NTG-induced relaxation (krelmax; p < 0.01) and desensitization (kdes; p < 0.03) responses was significantly greater in corporal tissue strips excised from diabetic than nondiabetic patients. Furthermore, the EC50for NTG-induced relaxation of precontracted corporal smooth muscle strips from potent patients (≈25 nM) was 0.90 log unit less than that for equivalently contracted corporal smooth muscle strips derived from impotent patients (≈180 nM; p < 0.03). Such observations suggest that alterations in corporal smooth muscle responsivity to activation of the guanylate cyclase – cGMP pathway, per se, may be a characteristic of organic erectile dysfunction. In the absence of compensatory changes in other vasodilatory mechanisms, this may contribute to incomplete corporal smooth muscle relaxation and the etiology of erectile dysfunction in some patients.Key words: nitroglycerine, corporal smooth muscle, relaxation, desensitization, guanylate cyclase.
Similarities between vasoconstrictor- and veratridine-stimulated metabolism in perfused rat hind limb
Authors: Tong Alex CY , Rattigan Stephen , Clark Michael G
Published: April 2011

The vasoconstrictors norepinephrine (NE) and angiotensin II (AII) mediate increases in oxygen uptake by the constant flow perfused rat hind limb that are inhibited by quinidine-like membrane-stabilizing effects (involving the interruption of action potential) of ( ±)-propranolol with little effect on vasoconstriction. The membrane labilizer veratridine, 10 µM, which has the capability of maintaining voltage-gated Na+ channels of the plasma membrane in their open state, also increases oxygen uptake but without an increase in pressure. Thus in the present study veratridine was characterized in detail and compared with NE in the same system. Veratridine (3-100 µM) produced a dose-dependent stimulation of oxygen uptake (from 11.8 ± 0.3 to 20.4 ± 0.6 µmol ·h-1 ·g-1 (n = 5), p < 0.0001) and lactate efflux (LE) (from 7.4 ± 0.6 to 23.0 ± 4.7 µmol ·h-1 ·g-1 (n = 5), p < 0.01). These increases were independent of vasoconstriction at low doses ( <= 10 µM). At higher doses of veratridine the accompanying minor vasoconstriction (from 17 ± 1 to 30 ± 2 mmHg (1 mmHg = 133.3 Pa) (n = 5), p < 0.005) was blocked by sodium nitroprusside (NP) while neither oxygen uptake nor LE was greatly affected. Low Na+ perfusions (25 mM) did not affect the vasoconstrictor action of NE but markedly inhibited increases in oxygen uptake and LE due to either veratridine or NE. Veratridine (10 µM) mediated increases in oxygen uptake and LE were blocked by either ( ±)-propranolol (100 µM) or 150 µM quinidine. It is concluded that vasoconstrictors such as NE, which stimulate oxygen uptake in the perfused rat hind limb, do so by a two-stage process involving an essential nitroprusside-sensitive redirection of flow followed by a mechanism involving increased ion movement across skeletal muscle cell membranes, which is blocked by membrane stabilizers. Veratridine achieves a similar increase in oxygen uptake but may do so by directly destabilizing the skeletal muscle cell membrane without the requirement of a redirection of flow.Key words: membrane labilizer, vascular control of muscle metabolism, paracrine substances of vascular origin, Na+ channel.
Effect of chronic NG-nitro-L-arginine methyl ester (L-NAME) on blood pressure and renal function in conscious uninephrectomized spontaneously hypertensive rats
Authors: Reverte María , Flores Olga , Gallego Belén , Lestón Antonio , López-Novoa José Miguel
Published: April 2011

We have studied during 30 days the effect of a low dose of NG-nitro-L-arginine methyl ester (1 mg ·kg-1 ·day-1 in drinking water) in the presence of D- or L-arginine (1 mg ·kg-1 ·day-1 in drinking water) in comparison with D- or L-arginine alone on blood pressure and renal function in conscious uninephrectomized female spontaneously hypertensive rats. At the end of the study, there was a significant increase in systolic blood pressure in the NG-nitro-L-arginine methyl ester + D-arginine group (307 ± 6 mmHg (1 mmHg = 133.3 Pa), n = 14, p << 0.05) in comparison with NG-nitro-L-arginine methyl ester + L-arginine (281 ± 6 mmHg, n = 14), L-arginine (262 ± 5 mmHg, n = 13), and D-arginine (258 ± 7 mmHg, n = 12) groups. There were no changes in diuresis, proteinuria, or sodium and potassium excretion between differently treated animals during this study. These results suggest that in uninephrectomized female spontaneously hypertensive rats, after 1 month blockade of NO synthesis with a low dose of NG-nitro-L-arginine methyl ester, vasculature is under tonic control by NO and it is not correlated with renal dysfunction.Key words: Key words: NG -nitro-L-arginine methyl ester (L-NAME), kidney, hypertension, spontaneously hypertensive rats, renaldysfunction, uninephrectomy.
Electrical conduction within the cerebrovasculature of stroke-prone spontaneously hypertensive rats
Authors: Smeda John S , King Shelley R
Published: April 2011

Alterations in electrical conductivity between smooth muscle cells (SMCs) can alter the spread and effectiveness of electromechanical SMC contraction. We attempted to determine whether alterations in pressure-dependent constriction (PDC) occurring in relation to stroke development within the middle cerebral arteries (MCAs) of Wistar-Kyoto stroke-prone hypertensive rats (SHRsp) were associated with changes in electrical conductivity between the SMCs. Current was injected into nonpressurized MCAs, using a suction electrode. The conducting distance along the length of the MCA where the amplitude of the membrane potential deflection (electrotonic potential) produced by current injection declined to 1/e (length constant) was used to measure conductivity. PDC to a 100 mmHg pressure step was measured with a pressure myograph. A loss of PDC in the MCAs of SHRsp preceded stroke development. Heptanol (4 mM), a gap junction communication inhibitor, reversibly inhibited conductivity and PDC in the MCA of prestroke SHRsp. The ability of heptanol to reversibly inhibit PDC was likely not related to it's ability to alter electrical conduction. The length constant of electrical conduction in the MCAs was about 0.75 mm and didn't differ between MCA sampled from pre- versus post-stroke SHRsp or Sprague-Dawley rats. It was concluded that alterations in electrical conductivity along the MCA could modify the spread of PDC, but such changes do not contribute to the loss of PDC within the MCA of poststroke SHRsp.Key words: membrane potential, electrotonic potential, middle cerebral arteries, myogenic response, gap junction, stroke-prone hypertensive rats.
The study of a physiological significance of prolactin in the regulation of calcium metabolism during pregnancy and lactation in rats
Authors: Lotinun Sutada , Limlomwongse Liangchai , Krishnamra Nateetip
Published: April 2011

Since a pharmacological dose of prolactin has previously been reported to enhance calcium absorption and bone calcium turnover, the role of endogenous prolactin in the regulation of calcium metabolism was investigated in the balance studies of Wistar rats between days 17 and 20 of first (P1) and fourth (P4) pregnancy and between days 12 and 15 of lactation (L). Each group was divided into 3 subgroups: one subgroup was given 0.9% NaCl (control); one was given 0.3 mg bromocriptine/100 g body weight ip twice daily for 3 days (to suppress prolactin secretion); and one was given bromocriptine and 0.25 mg prolactin/100 g body weight sc daily for 3 days. All three groups received 1 mL/100 g body weight of 1.25 mM calcium gluconate containing 2 mCi (1 Ci = 37 GBq) 45Ca daily for 3 days. Compared with the two pregnant controls, the L group had higher food consumption and higher fecal calcium excretion and lower urinary calcium excretion (% intake). Bromocriptine administration increased total calcium excretion from 59% intake to 84 and 66% intake in P1 and P4, respectively, suggesting that endogenous prolactin decreased total calcium excretion. On the other hand, exogenous prolactin had no effect on the calcium balance of P1 but increased the total calcium excretion in P4 from 57 to 66% intake. In contrast, the calcium balance of lactating rats was not altered by suppression of endogenous prolactin secretion or exogenous prolactin. Considering bone 45Ca content as representing bone Ca turnover, a lower value of bone 45Ca content indicated an accelerated bone Ca turnover. It was found that bromocriptine had no effect in P1 but decreased bone Ca turnover rate in the P4 and L groups, indicating an accelerating effect of endogenous prolactin on bone Ca turnover in the P4 and L groups. Exogenous prolactin, on the other hand, decreased bone Ca turnover rate in every group. Muscle Ca turnover was affected by bromocriptine and exogenous prolactin in the same manner as bone 45Ca contents. Interestingly, the biphasic action of prolactin was demonstrated in both calcium absorption and bone calcium turnover. It could be concluded that during pregnancy and lactation, endogenous prolactin increases food consumption, fractional calcium absorption, and bone calcium turnover, apparently to increase calcium availability for fetal development and milk calcium secretion.Key words: bone turnover, bromocriptine, calcium absorption, calcium excretion, calcium retention, prolactin.
Conditioned responses elicited by experimentally produced cues for smoking
Authors: Mucha R F , Pauli P , Angrilli A
Published: April 2011

Several theories of drug-craving postulate that a signal for drug elicits conditioned responses. However, depending on the theory, a drug cue is said to elicit drug similar, drug compensatory, positive motivational, and negative motivational effects. Since animal data alone cannot tease apart the relative importance of different cue-related processes in the addict, we developed and examined a model of drug cues in the human based on a two-sound, differential conditioning procedure using smoking as the reinforcer. After multiple pairings of a sound with smoking, there was a preference for the smoking cue on a conditioned preference test. The acute effects of smoking (increased heart rate, respiration rate, skin conductance level, skin conductance fluctuations, EEG beta power and trapezius EMG, decreased alpha power) were not affected by the smoking cue, although subjects drew more on their cigarette in the presence of the smoking cue than in the presence of a control cue. Moreover, the cue did not change baseline behaviour except for a possible increase in EEG beta power and an increase in trapezius EMG at about the time when smoking should have occurred. The findings confirm the value of experimental models of drug cues in the human for comparing different cue phenomena in the dependent individual. They indicate that an acquired signal for drug in the human may elicit incentive motivational effects and associated preparatory motor responses in addition to possible conditioned tolerance.Key words: drug cues, conditioning, smoking, tolerance, human.
Pharmacological studies on the "orphan" opioid receptor in central and peripheral sites
Authors: Nicholson J R , Paterson S J , Menzies JRW , Corbett A D , McKnight A T
Published: April 2011

We have exploited the availability of the "orphan" opioid receptor (referred to here as ORL1) in its "natural state" to investigate the effect of nociceptin (orphanin FQ), the endogenous agonist for the ORL1receptor in the brain, vas deferens, and myenteric plexus of the small intestine. Nociceptin was a potent agonist in electrically stimulated preparations of vasa deferentia (rat and rabbit) and myenteric plexus (guinea-pig) (IC50ranging from 18 to 31 nM) and susceptible to enzymic cleavage as addition of a cocktail of peptidase inhibitors to the organ bath produced a leftward shift in concentration-response curves (IC50ranging from 2.1 to 4.9 nM). In radioligand binding experiments using brain membranes from rat, rabbit, and guinea-pig, [3H]nociceptin bound a single population of binding sites with high affinity (KDvalues ranging from 0.049 to 0.124 nM) and capacity (Bmaxranging from 143 to 254 fmol ·mg-1protein). However, the response to nociceptin in functional studies and in radioligand binding inhibitory assays was resistant to antagonism/displacement by naloxone and a range of other opioid receptor antagonists, thus displaying a very different pharmacological profile from that of the "classical" opioids. Therefore, we conclude that the effect of nociceptin in these studies is not via an action at µ, delta , or kappa opioid receptors but rather at an orphan opioid receptor, ORL1.Key words: nociceptin, orphanin FQ, vas deferens, myenteric plexus, "orphan" opioid receptor.
The 1997 Stevenson Award Lecture. Cardiac K+channel gating: cloned delayed rectifier mechanisms and drug modulation
Authors: Fedida David , Chen Fred SP , Zhang Xue
Published: April 2011

K+channels are ubiquitous membrane proteins, which have a central role in the control of cell excitability. In the heart, voltage-gated delayed rectifier K+channels, like Kv1.5, determine repolarization and the cardiac action potential plateau duration. Here we review the broader properties of cloned voltage-gated K+channels with specific reference to the hKv1.5 channel in heart. We discuss the basic structural components of K+channels such as the pore, voltage sensor, and fast inactivation, all of which have been extensively studied. Slow, or C-type, inactivation and the structural features that control pore opening are less well understood, although recent studies have given new insight into these problems. Information about channel transitions that occur prior to opening is provided by gating currents, which reflect charge-carrying transitions between kinetic closed states. By studying modulation of the gating properties of K+channels by cations and with drugs, we can make a more complete interpretation of the state dependence of drug and ion interactions with the channel. In this way we can uncover the detailed mechanisms of action of K+channel blockers such as tetraethylammonium ions and 4-aminopyridine, and antiarrhythmic agents such as nifedipine and quinidine.Key words: potassium channel, Kv1.5, channel gating, inactivation, pore region, gating currents.
Ventilatory and metabolic effects of hypercapnia in conscious rats: AVP V1 receptor block
Authors: Walker Julia KL , Jennings Donald B
Published: April 2011

In conscious dogs, arginine vasopressin (AVP) inhibits an angiotensin II drive to ventilation during air breathing and during acute hypercapnia. To determine whether AVP inhibits respiration in rats, as in dogs, respiration and metabolism were measured in six male Sprague-Dawley rats using a plethysmograph. Rats breathed air, followed by 5% and 6.5% CO2 with or without AVP V1 receptor block. In unblocked experiments, minute ventilation increased to a comparable level during inhalation of both CO2 gas mixtures, resulting in a flattening of the ventilatory response to increased PaCO2. However, oxygen consumption decreased during 6.5% CO2, compared with 5% CO2, so that the ventilatory equivalent for O2 increased in a more linear manner with respect to PaCO2. The main effect of AVP V1 receptor block was to increase mean arterial blood pressure; there was no significant effect of AVP V1 receptor block on respiratory responses. AVP does not inhibit respiration in conscious rats as it does in conscious dogs.Key words: respiratory control, respiratory pattern, inspiratory and expiratory times, ventilatory equivalent for oxygen.
Effect of vasoactive intestinal peptide on pacemaker location and heart rate in the dog atrium
Authors: Pintér Arnold , Nadeau Réginald , Dandan Nazih , Pagé Pierre L
Published: April 2011

Vasoactive intestinal polypeptide (VIP) was either injectedintravenously (300 pmol·kg1) or perfused (1 nmol in 1 min) into the sinusnode artery (SNA) in anesthetized dogs to study its effect on subsidiary atrial pacemakers.Isochronal maps were obtained from 128 unipolar electrograms recorded on the epicardialsurface of both atria in nine animals. When VIP was perfused into the SNA or injectedintravenously, heart rate increased by 29 ± 16% and 12 ± 12%, and blood pressure decreased by16 ± 15 mmHg (1 mmHg = 133.3 Pa) and 24 ± 18 mmHg, respectively. Nosignificant change in heart rate (3 ± 6% decrease) accompanied a similar decrease in bloodpressure after an intravenous sodium nitroprusside perfusion. The perfusion of VIP into the SNAas well as the intravenous injection of VIP induced a shift of the pacemaker site to the region ofBachmanns bundle in a third of the preparations, while the pacemaker remained in the sinusnode area in two thirds. A perfusion of isoproterenol into the SNA produced a similar heart rateincrease (32 ± 14%, NS vs. VIP), and shifted the pacemaker site rostrally within the sinus node inthree of five preparations, or to the region of Bachmanns bundle in two of five preparations.The response to VIP in the location of the pacemaker was significantly different from theresponse to isoproterenol. Repeated perfusions of VIP into the SNA after 10-, 25-, 40-, and60-min intervals produced 2 ± 13% (p < 0.005 vs. the effect of first VIP administration),14 ± 12% (p < 0.05), 10 ± 12% (p < 0.05) and 30 ± 13% (NS) heart rateincreases, respectively, thereby demonstrating a tachyphylactic effect. In conclusion, VIP seemsto exert its positive chronotropic effect directly (probably via specific VIP receptors), althoughthe phenomenon of tachyphylaxis may suggest an indirect sympathomimetic mechanism.Key words: vasoactive intestinal peptide,subsidiary atrial pacemakers, sinus node artery, atrial mapping.
Insulin-sparing effect of hydroxychloroquine in diabetic rats is concentration dependent
Authors: Emami Jaber , Gerstein Hertzel C , Pasutto Franco M , Jamali Fakhreddin
Published: April 2011

To study the effect of hydroxychloroquine (HCQ) on glucose and insulin homeostasis, healthy rats were dosed with160 mg·kg-1·day-1 of HCQ orally, and streptozocin-induced diabetic rats received 80, 120, and 160 mg·kg-1·day-1 of HCQ, while controls received normal saline. Ten days after treatment with HCQ, healthy animals were challenged intravenously with insulin or glucose, while diabetic rats were given only an i.v. injection of insulin. In healthy rats, the areas within and under the glucose concentration - time curve following insulin and glucose challenge were estimated. In diabetic animals, the areas under the curve for both the percent change in serum glucose from baseline (AUG) and the percent change in serum insulin from baseline (AUI) were used as pharmacodynamic end points. In healthy rats, HCQ did not influence fasting serum glucose concentrations or glycemic profiles following i.v. administration of glucose or insulin. In diabetic rats, AUG and AUI were increased dependent on blood HCQ concentrations. The normal homeostatic mechanisms responsible for insulin-glucose regulation may compensate for possible HCQ-induced reduction of insulin metabolism in healthy rats. The HCQ dose- or concentration-effect relationships for glucose and insulin were linear over the range of HCQ concentrations tested. It is concluded that HCQ significantly elevated insulin blood concentration resulting in reduced glucose levels in a concentration-dependent fashion in diabetic rats. HCQ may have therapeutic potential in the treatment of type I and type II diabetes.Key words: hydroxychloroquine, enantiomers, insulin, glucose, diabetes, pharmacokinetics.
Tyrosine kinase inhibitors block the glucocorticoid stimulation of prostaglandin endoperoxide H synthase expression in amnion cells
Authors: Zakar Tamas , Mijovic Jane E , Bhardwaj Damyanti , Olson David M
Published: April 2011

Human amnion cells in primary culture respond to glucocorticoids in a characteristic fashion by the increased expression of the inducible prostaglandin endoperoxide H synthase isoenzyme, PGHS-2. Since PGHS-2 induction by agonists generally involves tyrosine kinases, we examined the possibility that the glucocorticoid stimulation of PGHS-2 in the amnion cells is tyrosine kinase dependent. PGHS-2 expression was stimulated in confluent, serum-starved amnion cells with dexamethasone, and the effect of the tyrosine kinase inhibitors herbimycin A and tyrphostins AG126, AG1288, and A1 on enzyme activity induction was determined. All four inhibitors blocked the increase of PGHS activity in a concentration-dependent manner with IC50 values of 0.077 ± 0.05, 15.38 ± 5.14, 20.91 ± 3.1, and 29.77 ± 8.21 µM, respectively (mean ± SE, n = 4). Dexamethasone increased (approximately twofold) the tyrosine phosphorylation of 120-, 110-, and 77-kDa proteins in cell extracts, and herbimycin A selectively blocked the phosphorylation of the 110-kDa phosphoprotein. The stimulation of the steady-state level of PGHS-2 mRNA by dexamethasone was also inhibited by herbimycin A. These results suggest that glucocorticoids induce PGHS-2 expression in amnion cells with the involvement of tyrosine kinase(s). The role of tyrosine kinase dependent mechanisms in the control of amnion cell responsiveness to corticosteroids remains to be established.Key words: amnion, glucocorticoid, tyrosine kinase, prostaglandin H synthase.
Are prostaglandins involved in atrial natriuretic peptide mechanisms of cardiovascular control?
Authors: Bandali Karim S , Ackermann Uwe
Published: April 2011

Atrial natriuretic peptide (ANP) can excite cardiac nerve endings and invoke a decrease in arterial blood pressure and a reduction in renal sympathetic nerve activity. Our laboratory has previously demonstrated that this renal depressor reflex was invoked by systemic injection of ANP and not by the direct application of ANP to the epicardium, a major locus for vagal afferents. We now examine whether inhibition of prostaglandin synthesis impairs reflex responses that are normally associated with ANP injections. Renal sympathetic nerve activity, arterial blood pressure, and heart rate were recorded in anesthetized rats. Indomethacin was used to inhibit prostaglandin synthesis through the cyclooxygenase pathway. The ANP-mediated decrease in arterial blood pressure and renal sympathetic nerve activity, observed when prostaglandin synthesis was inhibited, did not differ significantly from the decreases observed in these parameters when prostaglandin synthesis was not inhibited. Heart rate remained unchanged. Our results suggest that the sympatho-inhibitory effects of ANP do not require prostaglandins as intermediary compounds.Key words: sympathetic nervous system, renal nerves, prostaglandins.
Mitochondrial membrane potential regulation is independent of c-fosexpression
Authors: Moorehead Roger A , Singh Gurmit
Published: April 2011

Tumour cells contain mitochondria with elevated membrane potentials compared with normal cells, and thus this feature provides a selective target for destroying tumour cells. To improve mitochondrial-based therapies, a better understanding of the factors involved in regulating mitochondria are required. Since v-fos overexpression has been shown to elevate mitochondrial membrane potentials in rat fibroblasts, we investigated whether the human homologue, c-fos, was also capable of regulating the mitochondrial membrane potential in cells. Rat fibroblasts transfected with the c-fos gene did not accumulate more rhodamine 123 (Rh123) nor did they retain this Rh123 for extended periods of time compared with their parental line. Moreover, there was no difference in survival following dequalinium chloride (Deca) treatment between transfectants and controls. Similarly, reduction of c-fos expression in rat fibroblasts did not significantly alter their mitochondrial membrane potential. In addition, human ovarian carcinoma cells, which overexpress the c-fos gene, did not accumulate more Rh123 nor were they hypersensitive to Deca compared with their parental line. In another human ovarian carcinoma cell line, selection of variants with lower mitochondrial membrane potential did not alter c-fos mRNA or protein levels. These data suggest that alterations in c-fos expression do not regulate the magnitude of the mitochondrial membrane potential.Key words: c-fos, mitochondria, membrane potential, rhodamine 123 (Rh123), lipophilic cations.
Hemodynamic and biochemical effects of 100% oxygen breathing in humans
Authors: Milone Stephen D , Newton Gary E , Parker John D
Published: April 2011

High concentrations of inspired oxygen have been reported to have significant hemodynamic effects that may be related to increased free radical production. If oxygen therapy increases free radical production, it may also modify hemodynamic responses to a nitric oxide donor. Twenty-nine healthy male volunteers were studied using randomized, double-blind, placebo-controlled, crossover designs to determine whether oxygen therapy is associated with hemodynamic and forearm vascular effects. We measured hemodynamic parameters and forearm vascular responses before and 1 h after exposure to 100% oxygen versus medical air. Plasma 8-iso-PGF2α and plasma vitamin C were measured to assess the biochemical effects of oxygen administration. Hemodynamic measurements were also made following the acute administration of sublingual nitroglycerin. Oxygen therapy caused no significant change in blood pressure, plasma 8-iso-PGF2α, or vitamin C. Oxygen did cause a significant reduction in heart rate and forearm blood flow, and an increase in peripheral vascular resistance. Oxygen caused no change in the hemodynamic response to nitroglycerin. Therefore, in healthy young adults, therapy with 100% oxygen does not affect blood pressure, despite causing an increase in vascular resistance, is not associated with evidence of increased free radical injury, and does not affect the hemodynamic responses to nitroglycerin.Key words: oxygen, nitroglycerin, free radicals, blood pressure, heart rate, plethysmography.
Heterogeneity among β-adrenoreceptor blockers in the modulation of energy-dependent uptake of the organic cation amantadine by rat renal tubules
Authors: Stupack Dwayne G , Escobar Miguel R , Carlisle Maxine , Davie Thomas , Sitar Daniel S
Published: April 2011

Eight representative β-adrenoreceptor blocking drugs, acebutolol, atenolol, labetalol, metoprolol, nadolol, pindolol, propranolol, and timolol, were studied in vitro with respect to their potential to block energy-dependent uptake of [3H]amantadine into proximal and distal rat renal tubule fragments in the presence and absence of bicarbonate. Five of the eight β-adrenoreceptor blockers showed a dose-dependent inhibition of renal tubule accumulation of amantadine: labetalol, metoprolol, pindolol, propranolol, and timolol. Labetalol was the only β-adrenoreceptor blocker with greater inhibitory potency in phosphate-based buffer than in bicarbonate-based buffer. Propranolol was the most potent inhibitor of renal tubule amantadine accumulation with IC50values of 15 ± 10 and 31 ± 11 µM for proximal and distal tubule fragments, respectively, in a bicarbonate-based buffer environment. Inhibition in a phosphate-based buffer was less potent only in proximal tubules, with an IC50of 76 ± 30 µM. Kinetic studies of propranolol inhibition of amantadine uptake were consistent with both uncompetitive and competitive inhibition mechanisms in bicarbonate-based buffer in both proximal and distal renal tubule segments. There was no chiral preference between the R and S forms of propranolol for the inhibitory effects observed. These data suggest that there is potential for selection among the β-adrenoreceptor blocking drugs to minimize or restrict the inhibition of amantadine energy-dependent uptake at the organic cation ion uptake sites characterized by amantadine in the presence and absence of bicarbonate.Key words: organic cation transport, kidney, amantadine, β-adrenoreceptor blockers, bicarbonate.
Lack of mechanism-based inactivation of rat hepatic microsomal cytochromes P450 by doxorubicin
Authors: Re Johnny Di , Lee Chunja , Riddick David S
Published: April 2011

Administration of the antineoplastic doxorubicin to rodents causes depression of hepatic cytochrome P450 (CYP) dependent biotransformation, an effect that has been partially attributed to the ability of doxorubicin to stimulate microsomal lipid peroxidation. Since doxorubicin can be bioactivated by the CYP/NADPH-CYP reductase system to products that bind covalently to microsomal protein, we hypothesized that doxorubicin functions as a mechanism-based inactivator of hepatic microsomal CYPs and (or) NADPH-CYP reductase under conditions in which doxorubicin-stimulated NADPH-dependent lipid peroxidation is minimized. In vitro studies were conducted with hepatic microsomes isolated from untreated and phenobarbital-treated male rats. Unlike the positive control carbon tetrachloride, doxorubicin (10 µM) did not stimulate NADPH-dependent lipid peroxidation in microsomal incubations containing EDTA (1.5 mM). Doxorubicin did not cause NADPH-dependent loss of microsomal CYP, heme, or steroid hydroxylation activities selective for CYP2A, CYP2B, CYP2C11, and CYP3A. The positive control 1-aminobenzotriazole caused marked NADPH-dependent decreases in all of these parameters. Neither doxorubicin nor 1-aminobenzotriazole caused NADPH-dependent loss of NADPH-CYP reductase activity, and neither compound altered the immunoreactive protein levels of CYP2B, CYP2C11, CYP3A, and NADPH-CYP reductase. These results indicate that a pharmacologically relevant concentration of doxorubicin does not cause direct mechanism-based inactivation of hepatic microsomal CYPs or NADPH-CYP reductase, suggesting that the ability of doxorubicin to depress hepatic CYP-mediated biotransformation in vivo is due to lipid peroxidation mediated heme destruction, altered heme metabolism, and (or) decreased expression of selected CYP enzymes.Key words: doxorubicin, cytochrome P450, mechanism-based inactivation, lipid peroxidation.
Bezafibrate, an anti-hypertriglyceridemic drug, attenuates vascular hyperresponsiveness and elevated blood pressure in fructose-induced hypertensive rats
Authors: Si Xiaochen , Webb R Clinton , Richey Joyce M
Published: April 2011

A high fructose diet induces hypertension, hyperinsulinemia - insulin resistance, and hypertriglyceridemia (syndrome X). In this study, we investigated the role of an abnormal lipid profile in mediating fructose-induced hypertension. We hypothesized that bezafibrate, a lipid-lowering drug, would reduce elevated blood pressure and inhibit increased vascular reactivity in fructose-fed rats. Male rats were placed on four different diets: group 1 was fed standard chow (n = 6); group 2 was fed 60% fructose (n = 5); group 3 was fed fructose plus bezafibrate (30 mg·kg-1·day-1; drinking water; n = 5); and group 4 was fed standard chow plus bezafibrate (n = 6). In addition, the direct effects of very low density lipoprotein (VLDL) on vascular reactivity were examined. Bezafibrate treatment lowered blood pressure, free fatty acids, and triglycerides in the fructose-fed group, suggesting that lipid abnormalities play a role in the elevation of blood pressure in the fructose-induced hypertensive rat. Aortae from fructose-fed rats were hyperresponsive to the calcium channel agonist Bay K 8644, which was normalized with bezafibrate treatment. Incubation of aortae in a VLDL medium resulted in increased responsiveness to Bay K 8644, lending further support to lipid abnormalities altering vascular reactivity. An altered lipid profile evidenced by elevated triglycerides and free fatty acids is causally related to the development of high blood pressure and increased vascular reactivity in the fructose-induced hypertensive rat.Key words: Sprague-Dawley rats, hypertriglyceridemia, free fatty acids, vascular reactivity, aortae.
Nonselective cationic currents activated by acetylcholine in swine tracheal smooth muscle cells
Authors: Yamashita Toshikazu , Kokubun Shinichiro
Published: April 2011

Membrane currents in isolated swine tracheal smooth muscle cells were investigated using a pipette solution containing BAPTA-Ca2+ buffer and Cs+ as the major cation. With a pipette solution containing 100 nM free Ca2+, acetylcholine (ACh; 1-100 µM), in a concentration-dependent manner, activated a current without inducing shortening of cells, although neither 1 mM histamine nor 1 µM leukotriene D4 activated the current (n = 7, n is the number of cells). The effect of 100 µM ACh was suppressed by pretreatment with 100 µM atropine (n = 6) or intracellular application of preactivated pertussis toxin at a concentration of 0.1 µg·mL-1 (n = 8). Genistein (0.1-100 µM), in a concentration-dependent manner, suppressed the activation of the inward current by 100 µM ACh, whereas it did not significantly suppress that of the outward current (n = 6-8). With a pipette solution containing 50 nM free Ca2+, outward current, but not inward current, was activated by 100 µM ACh (n = 10). When the pipette solution had free Ca2+ concentrations greater than 50 nM, the inward current together with the outward current was activated. The ratio between the amplitude of the inward and outward currents was significantly increased as the free Ca2+ concentration in the pipette solution increased. The steady-state activation curve of the ACh-activated current with the 50 nM free Ca2+ pipette solution was fitted by a single Boltzmann distribution (Vh = +69.8 mV, k = -11.9 mV, n = 10). The activation time constant became smaller as the membrane potential was more depolarized (164.3 ± 5.9 ms at +40 mV to 92.4 ± 6.3 ms at +120 mV, n = 10). The reversal potential was not significantly changed by reducing extracellular Cl- concentration to one-tenth of the control (n = 8), suggesting that the current is a nonselective cationic current. These results suggest that ACh activates an outward nonselective cationic current via pertussis toxin-sensitive G-protein(s) coupled with muscarinic receptors. Involvement of genistein-sensitive tyrosine kinase in the activation process of the current is unlikely.Key words: tracheal smooth muscle, nonselective cationic current, acetylcholine, Ca2+ dependency, genistein sensitivity.
Astrocyte-induced modulation of synaptic transmission
Authors: Araque Alfonso , Sanzgiri Rita P , Parpura Vladimir , Haydon Philip G
Published: April 2011

The idea that astrocytes simply provide structural and trophic support to neurons has been challenged by recent evidence demonstrating that astrocytes exhibit a form of excitability and communication based on intracellular Ca2+ variations and intercellular Ca2+ waves, which can be initiated by neuronal activity. These astrocyte Ca2+ variations have now been shown to induce glutamate-dependent Ca2+ elevations and slow inward currents in neurons. More recently, it has been demonstrated that synaptic transmission between cultured hippocampal neurons can be directly modulated by astrocytes. We have reported that astrocyte stimulation can increase the frequency of miniature synaptic currents. Furthermore, we also have demonstrated that an elevation in the intracellular Ca2+ in astrocytes induces a reduction in both excitatory and inhibitory evoked synaptic transmission through the activation of selective presynaptic metabotropic glutamate receptors.Key words: astrocyte-neuron signaling, glutamate receptors, calcium waves, neuronal electrical activity, synaptic transmission.
Hydrolysis of long-chain,n-3 fatty acid enriched chylomicrons by cardiac lipoprotein lipase
Authors: Levy Ryna , Herzberg Gene R
Published: April 2011

The hydrolysis of chylomicrons enriched in long-chain n-3 fatty acids by cardiac lipoprotein lipase was studied. In 60 min, 24.8% of the triacylglycerol fatty acids were released as free fatty acids. The fatty acids were hydrolyzed at different rates. DHA (docosahexaenoic acid, 22:6n-3) and EPA (eicosapentaenoic acid, 20:5n-3) were released at rates significantly less than average. Stearic acid (18:0), 20:1n-9, and alpha-linolenic acid (18:3n-3) were released significantly faster than average. There was no relationship between the rate of release of a fatty acid and the number of carbons or the number of double bonds. Lipoprotein lipase selectively hydrolyzes the fatty acids of chylomicron triacylglycerols. This selectively will result in remnants that are relatively depleted in 18:0, 20:1, and 18:3 and relatively enriched in 20:5 and 22:6.Key words: lipoprotein lipase, chylomicrons, fish oil, eicosapentaenoic acid, docosahexaenoic acid.
Dopamine D3 receptor binding by D3 agonist 7-OH-DPAT (7-hydroxy-dipropylaminotetralin) and antipsychotic drugs measured ex vivo by quantitative autoradiography
Authors: Kaichi Yasusuke , Nonaka Ryo-ichi , Hagino Yoko , Watanabe Masayuki
Published: April 2011

Because the dopamine D3 receptor is primarily expressed in regions of the limbic system of brain, it was proposed that it may represent a target for antipsychotic drugs that is free of extrapyramidal side effects. An ex vivo receptor binding technique employing [3H]7-OH-DPAT was used to evaluate in vivo occupancy of dopamine D3 receptors in the rat nucleus accumbens by selective D3 agonist 7-OH-DPAT (7-hydroxy-dipropylaminotetralin) and various antipsychotic drugs. With an ID50 value of 0.07 mg/kg, the selective D3 agonist (+)-7-OH-DPAT had the most potent inhibitory effect on ex vivo binding of [3H]7-OH-DPAT among all drugs tested. Clinical doses of phenothiazine drugs, such as chlorpromazine and levomepromazine, induce binding to D3 receptors in vivo, while atypical antipsychotic drugs, such as clozapine, pimozide, and sulpiride, are very weak in inhibiting ex vivo binding of [3H]7-OH-DPAT, indicating that the role of D3 receptors as targets of antipsychotic drugs free of extapyramidal side effects may not be important.Key words: dopamine D3 receptor, ex vivo autoradiography, antipsychotic drugs, 7-OH-DPAT (7-hydroxy-dipropylaminotetralin).
Three different vasoactive responses of rat tail artery to nicotine
Authors: Wang Rui , Wang Zunzhe
Published: April 2011

The vasoactive effects of nicotine on isolated rat tail artery tissues were studied. Nicotine transiently contracted rat tail artery tissues (EC50, 55.6 ± 2 µM) in an extracellular Ca2+ dependent and endothelium-independent fashion. The blockade of alpha1-adrenoceptors, but not alpha2-adrenoceptors or P2X purinoceptors, inhibited the nicotine-induced contraction by 38 ± 7% (p < 0.05). Nicotine (1 mM) depolarized membrane by 13 ± 3 mV, but did not affect L-type Ca2+ channel currents, of the isolated rat tail artery smooth muscle cells. The phenylephrine-precontracted tail artery tissues were relaxed by nicotine (EC50, 0.90 ± 0.31 mM), which was significantly inhibited after the blockade of nicotinic receptors. Simultaneous removal of phenylephrine and nicotine, after a complete relaxation of the phenylephrine-precontracted tail artery strips was achieved by nicotine at accumulated concentrations (>=10 mM), triggered a Ca2+-dependent rebound long-lasting vasoconstriction (n = 20). This rebound contraction was abolished in the absence of calcium or in the presence of tetracaine in the bath solution. Pretreatment of vascular tissues with a nicotinic receptor antagonist did not affect the nicotine-induced vasoconstriction or nicotine withdrawal induced rebound contraction. The elucidation of the triphasic vascular effects of nicotine and the underlying mechanisms is important for a better understanding of the complex vascular actions of nicotine.Key words: nicotine, smokeless tobacco, vascular smooth muscles, contraction, relaxation.
FMRFamides exert a unique modulation of rodent pancreatic polypeptide sensitive neuropeptide Y (NPY) receptors
Authors: Parker Steven L , Parker Michael S
Published: April 2011

FMRFamide and related peptides (RFamides) were found to inhibit the association binding of iodinated human pancreatic polypeptide ([125I]hPP) to Y5-like neuropeptide Y (NPY) receptor in rodent tissues. An allosteric regulation of the activity of the rodent kidney PP-sensitive neuropeptide Y (NPY) receptor by RFamides was indicated by potency decrease with particle concentration in the inhibition of the association binding of125I-labeled human pancreatic polypeptide (hPP) by RFamides at rabbit kidney membranes. The competition by C-terminal hexapeptide of hPP (LTRPRY.NH2) did not show such affinity change. The steady-state binding of hPP showed little sensitivity to any of the RFamides tested. The Y1-selective binding of [125I][Leu31,Pro34]hPYY (at 2 nM hPP) was much less sensitive to RFamides than the binding of [125I]hPP, albeit with some differences across tissue or cell types. The binding of Y2-selective agonist125I-labeled human peptide YY (3-36) was quite insensitive to RFamides. The presence of a unique component in the inhibition of hPP binding by RFamides was further indicated by a degree of antagonism with phospholipase C inhibitor U-73122, and by an only limited cooperation with a N5-amiloride compound, and with alkylator chloroethylclonidine. Change of the chirality of individual residues in the FMRFamide molecule produced a significant reduction of inhibitory potency only with D-Phe in the C-terminal position. Substitution of the (C-3) L-Met by L-Leu greatly increased the inhibitory potency of RFamides relative to otherwise identical congeners. RFamides could act both as ligands of membrane neighbors of the PP receptor, and as competitors of Y5-like NPY receptor epitopes that accommodate the C-terminal aspects of agonist peptides.Key words: Y1receptor, Y2receptor, Y5receptor, RFamide, allosteric interaction, hydrophobic pocket, amino acid chirality.
The trade-off between foraging activity and predation risk for Rana temporaria in different food environments
Authors: Eklöv Peter , Halvarsson Charlotta
Published: April 2011

We evaluated the trade-off between foraging activity and predation risk for larvae of an anuran species presented with different types of food resources. In the laboratory we examined the effects of benthic and phytoplankton resources and the two types combined on the activity and mortality of larvae of the common frog, Rana temporaria, exposed to predatory larva, Dytiscus marginalis. Predation mortality of tadpoles increased with the duration of the experiment and was highest in the presence of the phytoplankton resource alone. This was explained by a decrease in prey activity in the benthic- and combined-resource treatments when the predator was present, whereas in the phytoplankton treatment, the activity of the tadpoles were similar in the presence or absence of the predator. In the presence of the predator, prey mainly used the bottom of the aquarium in the benthic- and combined-resource treatments. In contrast, in the phytoplankton treatment they used the water column more than in the other treatments. In the presence of the predator, the prey had a lower consumption rate in the phytoplankton treatment than in the other treatments. Predator activity did not change among treatments. The results suggest that the trade-off between foraging activity and predation mortality for anuran larvae is strongly affected by the types of food resources present in the environment.