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1.
Foot Ankle Surg ; 21(2): 97-102, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25937408

ABSTRACT

BACKGROUND: Arthroscopic arthrodesis is an accepted technique for the ankle but less commonly reported for multiple hindfoot joints. We present a case series review to establish the feasibility of this surgery. METHODS: In a consecutive series of 112 arthroscopic hindfoot arthrodeses, 18 involved decortication and fusion of 2 or 3 hindfoot joints. These were reviewed to identify complications, union rates and improvement over preoperative state. RESULTS: Mean age was 62 (45-78). Mean follow-up was 4.4 (1.75-7.5) years. There were no perioperative complications. The four patients with triple fusions united with good or excellent outcomes. Of 14 patients with subtalar and talonavicular arthrodeses there were three talonavicular non-unions. CONCLUSIONS: Arthroscopic double and triple arthrodeses appear feasible salvage options for pain and deformity. Decortication of all three joints and rotatory correction of deformity is possible whilst preserving the soft tissue envelope. Late adjacent joint pain and arthrosis can occur.


Subject(s)
Arthritis/surgery , Arthrodesis/methods , Foot Joints/surgery , Aged , Arthroscopy , Feasibility Studies , Female , Humans , Male , Middle Aged
2.
Nature ; 444(7122): 1044-6, 2006 Dec 21.
Article in English | MEDLINE | ID: mdl-17183315

ABSTRACT

Gamma-ray bursts (GRBs) are known to come in two duration classes, separated at approximately 2 s. Long-duration bursts originate from star-forming regions in galaxies, have accompanying supernovae when these are near enough to observe and are probably caused by massive-star collapsars. Recent observations show that short-duration bursts originate in regions within their host galaxies that have lower star-formation rates, consistent with binary neutron star or neutron star-black hole mergers. Moreover, although their hosts are predominantly nearby galaxies, no supernovae have been so far associated with short-duration GRBs. Here we report that the bright, nearby GRB 060614 does not fit into either class. Its approximately 102-s duration groups it with long-duration GRBs, while its temporal lag and peak luminosity fall entirely within the short-duration GRB subclass. Moreover, very deep optical observations exclude an accompanying supernova, similar to short-duration GRBs. This combination of a long-duration event without an accompanying supernova poses a challenge to both the collapsar and the merging-neutron-star interpretations and opens the door to a new GRB classification scheme that straddles both long- and short-duration bursts.

3.
Eur J Pharmacol ; 382(3): 217-27, 1999 Oct 15.
Article in English | MEDLINE | ID: mdl-10556673

ABSTRACT

The muscarinic acetylcholine receptor antagonists, atropine and pirenzepine, produced an apparent insurmountable antagonism of muscarinic M(1) receptor-mediated intracellular Ca(2+) mobilization in Chinese hamster ovary (CHO) cells when tested against the agonists carbachol or xanomeline. Each antagonist caused a dextral shift of the agonist concentration-response curves with depression of the maximum response that was incomplete (i.e., saturated) and which varied with the pairs of agonist and antagonist. Equilibrium competition binding assays found no deviation from simple, reversible competitive behavior for either antagonist. The relative rates of dissociation of unlabeled atropine and pirenzepine were also assessed in radioligand kinetic studies and it was found that atropine dissociated from the receptor approximately 8-fold slower than pirenzepine. Numerical dynamic simulations suggested that the insurmountability of antagonism observed in the present study was probably a kinetic artifact related to the measurement of transient responses to a non-equilibrated agonist in the presence of a slowly dissociating antagonist. Importantly, the patterns of antagonism observed included a saturable depression of agonist maximal response, a mode of antagonism that is incompatible with the previously described phenomenon of hemi-equilibrium states. Monte Carlo simulations indicated that reasonable, semi-quantitative estimates of antagonist potency could be determined by a minor modification of standard methods, where equieffective agonist concentrations, rather than EC(50) values, are compared in the absence and presence of antagonist. Application of the latter approach to the functional data yielded estimates of antagonist potency that were in excellent agreement with those derived from the equilibrium binding assays, thus indicating that the present method can be useful for quantifying antagonist potency under non-equilibrium conditions.


Subject(s)
Atropine/pharmacology , Calcium/metabolism , Muscarinic Antagonists/pharmacology , Pirenzepine/pharmacology , Receptors, Muscarinic/drug effects , Animals , Artifacts , Binding, Competitive , CHO Cells , Carbachol/pharmacology , Cricetinae , Drug Antagonism , Humans , Intracellular Fluid/metabolism , Kinetics , Models, Biological , Monte Carlo Method , Muscarinic Agonists/pharmacology , Pyridines/pharmacology , Radioligand Assay , Receptor, Muscarinic M1 , Thiadiazoles/pharmacology , Time Factors
4.
J Pharmacol Exp Ther ; 289(3): 1220-8, 1999 Jun.
Article in English | MEDLINE | ID: mdl-10336509

ABSTRACT

Previous findings in our laboratory suggested that the M1 muscarinic acetylcholine receptor (mAChR) agonist xanomeline exhibits a novel mode of interaction that involves persistent binding to and activation of the M1 mAChR, subsequent to extensive washout, as well as a possible insurmountable element. In the present study, we examined this interaction in greater detail, using Chinese hamster ovary cells transfected with the genes for the M1 mAChR and neuronal nitric oxide synthase. Pretreatment of cells with xanomeline, followed by extensive washout, resulted in elevated basal levels of neuronal nitric oxide synthase activity that were suppressed by the antagonists atropine or pirenzepine in a concentration-dependent manner. Analysis of the data yielded estimates of Schild slope factors and pKB values for the antagonists that were consistent with a model of simple competition between these latter agents and the persistently bound form of xanomeline. The ability of the antagonists to produce parallel dextral shifts of the concentration-response curves to carbachol and xanomeline was also investigated. The interaction between xanomeline and pirenzepine appeared to be insurmountable, but this may have been due to an equilibrium artifact. In contrast, the interaction between xanomeline and atropine conformed to a model of competition, indicating that the mode of interaction of free xanomeline at the M1 mAChR is pharmacologically identical with that of the persistently bound form. Radioligand binding studies also showed that the presence of various concentrations of xanomeline had no significant effect on the calculated affinity of atropine or pirenzepine in inhibiting the binding of [3H]N-methylscopolamine. Overall, these findings suggest that the persistent attachment of xanomeline to the M1 mAChR does not prevent this agonist from interacting with the classic binding site in a competitive fashion.


Subject(s)
Muscarinic Agonists/pharmacology , Pyridines/pharmacology , Receptors, Muscarinic/physiology , Thiadiazoles/pharmacology , Animals , Atropine/pharmacology , Binding, Competitive , CHO Cells , Carbachol/pharmacology , Citrulline/metabolism , Cricetinae , Humans , Kinetics , N-Methylscopolamine/pharmacokinetics , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type I , Pirenzepine/pharmacology , Radioligand Assay , Receptor, Muscarinic M1 , Receptors, Muscarinic/genetics , Receptors, Muscarinic/metabolism , Recombinant Proteins/metabolism , Transfection , Tritium
5.
Neurochem Res ; 24(1): 85-94, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9973241

ABSTRACT

Regulation of nitric oxide (NO) formation is critical to ensure maintenance of appropriate cellular concentrations of this labile, signaling molecule. This study investigated the role exogenous and endogenously produced NO have in feeding back to regulate NO synthesis in intact cells. Two NO donors inhibited activation of neuronal NO synthase (nNOS) in response to the muscarinic receptor agonist carbachol in Chinese hamster ovary (CHO) cells stably transfected with the M1 muscarinic receptor and nNOS. The presence of the NO scavenger [2-(4-Carboxyphenyl)-4,4,5,5tetramethylimidazoline-1-oxyl-3-oxide potassium salt] (C-PTIO) potentiated carbachol-induced activation of nNOS in transfected CHO cells. C-PTIO also potentiated nNOS activity in response to the Ca2+ ionophore ionomycin. In contrast, the NO scavenger oxyhemoglobin depressed carbachol- and ionomycin-induced NO formation. These discrepant results suggest that it is unlikely that endogenously produced NO induces feed back inhibition at the level of nNOS activation itself. Exogenous sources of NO inhibited carbachol-induced inositol phosphates formation. However, endogenously produced NO did not appear to feed back to regulate phosphoinositide hydrolysis as there was no difference in [3H]inositol phosphates formation between cells that do or do not express nNOS. There was also no change in carbachol-induced [3H]inositol phosphates formation in the presence or absence of a NOS inhibitor or the NO scavenger C-PTIO. A decrease in the carbachol-mediated transient Ca2+ peak was observed in cells that express nNOS as compared to cells lacking the enzyme, suggesting that endogenous NO might inhibit receptor mediated Ca2+ signaling. This conclusion, however, was not supported by the lack of ability of a NOS inhibitor to modulate carbachol-induced Ca2+ elevations. Taken together, these results highlight differences in the regulation of the nNOS activation cascade by endogenous vs. exogenous sources of NO.


Subject(s)
Carbachol/pharmacology , Nitric Oxide Donors/pharmacology , Nitric Oxide Synthase/metabolism , Nitric Oxide/physiology , Receptors, Muscarinic/physiology , Signal Transduction/physiology , Animals , Benzoates/pharmacology , CHO Cells , Calcium/metabolism , Cricetinae , Enzyme Activation , Feedback , Humans , Imidazoles/pharmacology , Inositol Phosphates/metabolism , Ionomycin/pharmacology , Kinetics , Nitric Oxide/pharmacology , Nitric Oxide Synthase Type I , Nitroprusside/pharmacology , Penicillamine/analogs & derivatives , Penicillamine/pharmacology , Phosphatidylinositols/metabolism , Receptor, Muscarinic M1 , Receptors, Muscarinic/drug effects , Recombinant Proteins/drug effects , Recombinant Proteins/metabolism , S-Nitroso-N-Acetylpenicillamine , Signal Transduction/drug effects , Transfection
6.
J Neurochem ; 71(2): 487-97, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9681438

ABSTRACT

This study shows that activation of M1 muscarinic receptors, when coexpressed in Chinese hamster ovary (CHO)-K1 cells with neuronal nitric oxide (NO) synthase (nNOS), produces early and late phases of elevation of both intracellular Ca2+ concentration and nNOS activity. We examined the relationship between receptor-mediated increases in intracellular Ca2+ concentration and activation of nNOS over both short and long intervals using guanosine 3',5'-cyclic monophosphate (cGMP) formation as a measure of nNOS activity. The rapid phase of nNOS activation was dependent on release of Ca2+ from intracellular stores in both the CHO M1/nNOS transfected cells and in neuroblastoma (N1E-115) cells, in which muscarinic receptors and nNOS are endogenously expressed. Two single point mutations in the M1 muscarinic receptor that have previously been shown to uncouple differentially the receptor from phosphoinositide hydrolysis produced parallel attenuation of the rapid phase of nNOS activation. Characterization of the prolonged phase of nNOS activation was done using the conversion of L-[3H]arginine to L-[3H]citrulline as well as cGMP formation following stimulation of M1 muscarinic receptors for 60 min. Both responses were dependent on influx of extracellular Ca2+ and were accompanied by prolonged formation of NO at functionally effective levels as late as 60 min following receptor activation. Therefore, this study demonstrates for the first time the existence of two mechanistically distinct phases of nNOS activation that are dependent on different sources of Ca2+.


Subject(s)
Calcium/pharmacokinetics , Nitric Oxide Synthase/metabolism , Receptors, Muscarinic/genetics , Receptors, Muscarinic/metabolism , Acetylcholine/metabolism , Animals , CHO Cells , Calcium/metabolism , Carbachol/pharmacology , Chelating Agents/pharmacology , Citrulline/biosynthesis , Citrulline/metabolism , Cricetinae , Cyclic GMP/metabolism , Dinucleoside Phosphates/pharmacology , Egtazic Acid/pharmacology , Enzyme Activation/genetics , Humans , Kinetics , Muscarinic Agonists/pharmacology , Mutagenesis/physiology , Nerve Tissue Proteins/metabolism , Nitric Oxide Synthase Type I , Rats , Receptor, Muscarinic M1 , Signal Transduction/drug effects , Signal Transduction/physiology , Transfection , Tritium
7.
Synapse ; 26(3): 235-42, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9183813

ABSTRACT

This study investigated second messengers formed in response to calcitonin gene-related peptide (CGRP) in primary cultures of neonatal rat spinal cord. CGRP increased the level of cAMP above basal levels (50 pmol/mg protein) over a large range of concentrations. The concentration-response curve had an intermediate plateau at 180 pmol cAMP/mg protein in response to 0.01-0.1 nM CGRP and a maximal plateau of 850 pmol cAMP/mg protein at 300 nM CGRP. The biphasic concentration-response curve (EC50S of 0.7 pM and 22 nM) suggests activation of high- and low-affinity receptors for CGRP. Both neurons and nonneuronal cells contributed to the increase in cAMP formation in response to CGRP. The CGRP receptor blocker, CGRP8-37, inhibited the response to both 1 and 100 nM CGRP, providing additional support for the hypothesis that both high- and low-affinity receptors mediate the formation of cAMP. Only a high concentration of CGRP (1 microM) increased the formation of cGMP, and CGRP had no effect on the formation of inositol phosphates at any of the concentrations tested (0.1-1 microM). These results suggest that CGRP-induced responses in the spinal cord are mediated predominately via the formation of cAMP. The observation that both neurons and nonneuronal cells responded to CGRP indicate that this peptide may have multiple actions in the spinal cord.


Subject(s)
Animals, Newborn/metabolism , Calcitonin Gene-Related Peptide/pharmacology , Second Messenger Systems/physiology , Spinal Cord/metabolism , Animals , Calcitonin Gene-Related Peptide/antagonists & inhibitors , Cells, Cultured , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Inositol Phosphates/metabolism , Neuroglia/drug effects , Neuroglia/metabolism , Neurons/drug effects , Neurons/metabolism , Rats , Rats, Sprague-Dawley , Second Messenger Systems/drug effects , Signal Transduction/drug effects , Signal Transduction/physiology , Spinal Cord/drug effects
8.
Pharmacology ; 55(1): 10-7, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9309796

ABSTRACT

We investigated the coupling of the M2 muscarinic acetylcholine receptors expressed in Chinese hamster ovary cells to activation of neuronal nitric oxide (NO) synthase. Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an indirect measure of the generation of NO in Chinese hamster ovary cells. The muscarinic agonist carbachol induced marked time- and concentration-dependent enhancement of the activity of NO synthase. Activation of neuronal NO synthase by M2 muscarinic receptors was associated with a small increase in the concentration of intracellular Ca2+. These data suggest the presence of alternate mechanisms of activation of neuronal NO synthase which might be operative in the absence of large changes in the concentration of cellular Ca2+. These findings help to understand the mechanisms of activation of NO synthase.


Subject(s)
Calcium/metabolism , Neurons/enzymology , Nitric Oxide Synthase/biosynthesis , Receptors, Muscarinic/metabolism , Animals , CHO Cells/drug effects , CHO Cells/metabolism , Carbachol/pharmacology , Cricetinae , Cyclic GMP/biosynthesis , Cytosol/enzymology , Dose-Response Relationship, Drug , Enzyme Activation , Gene Expression Regulation, Enzymologic/drug effects , Gene Expression Regulation, Enzymologic/genetics , Muscarinic Agonists/pharmacology , Nitric Oxide/biosynthesis , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase/metabolism , Receptor, Muscarinic M2 , Transfection , Tumor Cells, Cultured/metabolism
9.
Brain Res ; 739(1-2): 263-75, 1996 Nov 11.
Article in English | MEDLINE | ID: mdl-8955947

ABSTRACT

Hyperalgesia is a characteristic of inflammation and is mediated, in part, by an increase in the excitability of spinal neurons. Although substance P does not appear to mediate fast synaptic events that underlie nociception in the spinal cord, it may contribute to the hyperalgesia and increased excitability of spinal neurons during inflammation induced by complete Freund's adjuvant. We examined the role of endogenous substance P in changes in the excitability of spinal neurons during adjuvant-induced, peripheral inflammation by determining the effect of a selective NK1 receptor antagonist (RP67580) on the nociceptive flexor reflex in adult rats. Experiments were conducted 2 or 3 days after injection of adjuvant. Animals exhibited moderate thermal hyperalgesia at this time. The flexor reflex was evoked by electrical stimulation of the sural nerve and was recorded in the ipsilateral hamstring muscles. The flexor reflex ipsilateral to the inflamed hindpaw was enhanced approximately two-fold compared to the flexor reflex evoked in untreated animals as determined by the number of potentials and the duration of the reflex. The enhanced reflex in adjuvant-treated animals was most likely due to an increase in the excitability of spinal interneurons because short-latency activity in the hamstring muscles did not differ between untreated animals and adjuvant-treated animals following electrical stimulation of the L5 dorsal root or the nerve innervating the muscle with a stimulus that was 1.3-1.5 times the threshold for excitation of A-fibers. Intrathecal administration of RP67580 (2.3 and 6.8 nmol) attenuated the flexor reflex evoked in adjuvant-treated animals, but had no effect in untreated animals. Intravenous or intraplantar injection of RP67580 (6.8 nmol) did not affect the flexor reflex in adjuvant-treated animals indicating a spinal action of the drug following intrathecal administration. RP68651, the enantiomer of RP67580, was without effect at doses up to 6.8 nmol, indicating that the effects of comparable doses of RP67580 were due to an action of the drug at NK1 receptors. However, intrathecal administration of 23 nmol of both drugs attenuated the reflex in adjuvant-treated and control animals indicating that effects of RP67580 at this dose were not mediated entirely by its action at NK1 receptors. Overall, these data suggest that endogenous substance P has a role in the increased excitability of spinal interneurons observed during persistent inflammation and support the hypothesis that substance P released in the spinal cord contributes to the hyperalgesia that accompanies adjuvant-induced persistent, peripheral inflammation.


Subject(s)
Motor Neurons/physiology , Neuritis/physiopathology , Nociceptors/physiology , Receptors, Neurokinin-1/physiology , Reflex, Stretch/physiology , Spinal Cord/physiology , Afferent Pathways/drug effects , Afferent Pathways/physiology , Animals , Chronic Disease , Electric Stimulation , Electromyography , Hyperalgesia/physiopathology , Male , Motor Neurons/drug effects , Neuromuscular Junction/drug effects , Neuromuscular Junction/physiology , Nociceptors/drug effects , Rats , Rats, Sprague-Dawley , Receptors, Neurokinin-1/drug effects , Reflex, Stretch/drug effects , Spinal Cord/cytology , Spinal Cord/drug effects , Sural Nerve/physiology
10.
Neuroscience ; 68(3): 855-65, 1995 Oct.
Article in English | MEDLINE | ID: mdl-8577379

ABSTRACT

The naturally occurring tachykinins, substance P, neurokinin A and neurokinin B, induce the formation of inositol phosphates or cAMP in a variety of tissues but their effects on neurons have not been resolved. We used primary cultures of neonatal rat spinal cord to determine whether neurokinin receptors mediate changes in these second messengers in spinal neurons. We found that substance P, neurokinin A and neurokinin B induced the formation of inositol phosphates in a concentration-dependent manner with similar potencies (EC50S: 3.6, 5.7 and 21.3 nM, respectively), but at concentrations tested (0.1-1.0 microM) these peptides had no effect on cAMP levels. All three tachykinins induced the formation of inositol phosphates predominately by activation of neurokinin1 receptors. CP-96,345 and WIN 51,708, neurokinin1 receptor antagonists, attenuated the response to substance P, neurokinin A and neurokinin B. GR 103,537, a neurokinin2 receptor antagonist, had no effect on the responses induced by any of the tachykinins. Furthermore, the selective neurokinin1 receptor agonist, GR-73632, induced the formation of inositol phosphates in a concentration-dependent manner, whereas the selective neurokinin2 receptor agonist, GR-64349, generated inositol phosphates only at the highest concentration tested (10 microM). Senktide, a neurokinin3 receptor agonist, did not induce the formation of inositol phosphates at any of the concentrations tested (0.01-10 microM). Inositol phosphate formation appeared to be due to a direct effect of the tachykinins on neuronal neurokinin1 receptors. These results suggest that biological responses in spinal neurons following activation of neurokinin1 receptors are mediated mainly by the hydrolysis of phosphoinositol 4,5-bisphosphate to form inositol 1,4,5-trisphosphate and diacylglycerol. It remains to be determined which of these second messengers mediates the increased neuronal excitability and depolarization that occurs in response to substance P.


Subject(s)
Cyclic AMP/metabolism , Inositol Phosphates/biosynthesis , Neurons/metabolism , Receptors, Neurokinin-1/drug effects , Receptors, Neurokinin-2/drug effects , Spinal Cord/metabolism , Tachykinins/pharmacology , Animals , Animals, Newborn , Colforsin/pharmacology , Female , Neurokinin-1 Receptor Antagonists , Neurons/drug effects , Pregnancy , Rats , Rats, Sprague-Dawley , Receptors, Neurokinin-1/agonists , Receptors, Neurokinin-2/agonists , Receptors, Neurokinin-2/antagonists & inhibitors , Spinal Cord/cytology , Spinal Cord/drug effects , Synaptic Transmission/drug effects , Type C Phospholipases/metabolism
11.
J Pharmacol Exp Ther ; 261(3): 1206-12, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1376357

ABSTRACT

The neuropeptide substance P (SP) produces transient elevations in short-circuit current (Isc), a measure of active ion transport, across sheets of small intestinal mucosae from several animal species, but the ionic basis of this action remains unknown. The aim of this study was to test the hypothesis that SP promotes electrogenic anion secretion in the porcine proximal jejunum, an intestinal segment analogous to the human upper small intestine. Sheets of jejunal mucosa with attached submucosa responded to serosal (S), but not luminal (L) addition of 0.1 microM SP with a transient increase in Isc that was reduced in tissues pretreated with the Na(+)-K(+)-Cl- cotransport inhibitor bumetanide (10 microM) or bathed in media lacking Cl- or HCO3- ions. SP produced biphasic effects on transepithelial Na+ and Cl- fluxes; it initially stimulated a L-directed Na+ secretory flux during a 5-min period in which peptide-induced Isc elevations were maximum. The return of the Isc to base-line levels was temporally associated with an increase in L-directed Cl- transport. Both effects of SP were absent in tissues either pretreated with the neuronal conduction blocker tetrodotoxin (0.1 microM) or bathed in HCO3(-)-deficient media. Bumetanide abolished the Na+ secretory actions of SP, but did not affect peptide-induced Cl- secretion. pH-Stat titration experiments revealed that mucosal sheets alkalinized the L bathing medium at a rate twice that of the S medium. SP simultaneously increased and suppressed L and S alkalinization, respectively; this effect presumably represents HCO3- secretion.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Bicarbonates/metabolism , Jejunum/metabolism , Sodium/metabolism , Substance P/pharmacology , Animals , Biological Transport, Active/drug effects , Bumetanide/pharmacology , Chlorides/metabolism , Female , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Jejunum/drug effects , Male , Sensory Receptor Cells/drug effects , Sensory Receptor Cells/metabolism , Swine
12.
J Pharmacol Exp Ther ; 261(3): 1213-21, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1376358

ABSTRACT

Neurokinin (NK) peptides such as substance P (SP) may modulate epithelial ion transport in the small intestine. The present study was undertaken to examine the pharmacological mechanisms by which SP and its endogenous homologs NKA and NKB affect active electrolyte transport in the mucosa of porcine jejunum. Neurokinins and NK agonist analogs increased short circuit current, a measure of active ion transport, across sheets of jejunal mucosa-submucosa with the order of potency: SP greater than [beta-Ala8]NKA4-10 greater than or equal to [Sar9,Met(O2)11]SP greater than NKA = Arg-NKB greater than NKB after their addition to the serosal aspect of tissues (SP EC50 = 11 nM). Epithelial responses to SP or NKA underwent rapid autotachyphylaxis and unidirectional cross-tachyphylaxis after repeated peptide administration. The neuronal conduction blocker tetrodotoxin significantly reduced NK efficacy. SP activity was significantly reduced in tissues pretreated with the muscarinic cholinoceptor blocker atropine or the eicosanoid synthesis inhibitor eicosa-5,8,11,14-tetraynoic acid. NK peptides increased contractility in longitudinally oriented strips of jejunal smooth muscle with an order of potency: [Sar9,Met(O2)11]SP greater than SP greater than Arg-NKB = [beta-Ala8]NKA4-10 greater than or equal to NKB = NKA (SP EC50 = 11 nM). SP-induced contractions were reduced by 70 to 80% in tissues pretreated with atropine or the neuronal Ca++ channel blocker, omega-conotoxin. [125I]Bolton-Hunter-substance P (BHSP) bound specifically to a single population of sites in slide-mounted sections of mucosa-submucosa and smooth muscle with Kd = 0.3 and 0.1 nM and Bmax = 18 and 31 fmol/mg protein, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Jejunum/drug effects , Neurokinin A/pharmacology , Neurokinin B/pharmacology , Receptors, Neurotransmitter/drug effects , Substance P/pharmacology , Animals , Autoradiography , Binding Sites , Biological Transport, Active , Female , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Jejunum/metabolism , Male , Muscle Contraction/drug effects , Neurokinin A/metabolism , Neurokinin B/metabolism , Receptors, Neurokinin-2 , Substance P/metabolism , Succinimides/metabolism , Swine
13.
Calcif Tissue Int ; 47(6): 383-7, 1990 Dec.
Article in English | MEDLINE | ID: mdl-2292082

ABSTRACT

Female Sprague-Dawley rats (n = 72), 6 months old, underwent either sham operation, oophorectomy, adrenalectomy, or combined oophorectomy and adrenalectomy (O&A). They were all maintained on normal saline ad libitum and 20 g/day 1.1% calcium chow. Nine weeks after operation, the trabecular bone volume of the distal femoral shaft was significantly lower (P less than 0.001) in the adrenalectomized (11.1%), oophorectomized (7.0%), and O&A (8.3%) animals than in sham-operated animals (19.8%). Eighteen weeks after operation, the trabecular bone volume in O&A animals had fallen to a mean of 3.8% (sham 17.0%), and the length of the femur had increased to 38.8 mm after O&A (sham 36.8 mm, P less than 0.01). O&A animals treated with 0.35 mg/kg/week nandrolone decanoate from 9 weeks postoperatively onward, had twice the femoral trabecular bone volume of untreated animals at 18 weeks (P less than 0.05). By contrast, no significant differences were found in vertebral body trabecular bone between any groups, including groups receiving treatment with androgens. We have found that, by 9 weeks after operation, adrenalectomy alone causes significant loss of metaphyseal trabecular bone, similar to the progressive loss seen after oophorectomy.


Subject(s)
Adrenalectomy , Osteoporosis/etiology , Ovariectomy , Adrenal Glands/metabolism , Adrenal Glands/physiology , Adrenal Glands/surgery , Animals , Bone Density/drug effects , Bone Diseases, Metabolic/etiology , Bone Diseases, Metabolic/metabolism , Bone and Bones/drug effects , Bone and Bones/metabolism , Bone and Bones/physiology , Female , Minerals/metabolism , Nandrolone/pharmacology , Osteoporosis/metabolism , Osteoporosis/physiopathology , Ovary/metabolism , Ovary/physiology , Ovary/surgery , Rats , Rats, Inbred Strains
14.
Peptides ; 11(4): 779-87, 1990.
Article in English | MEDLINE | ID: mdl-2172944

ABSTRACT

The biochemical characteristics of specific binding sites for gastrin releasing peptide were examined in a preparation of the mucosa and submucosa of the porcine jejunum. Iodinated gastrin releasing peptide bound to sites in this preparation with an affinity that was similar to its potency in stimulating anion transport in the epithelium. The relative order of potency of peptide analogues of gastrin releasing peptide in contracting longitudinal smooth muscle was similar to their order of potency in altering anion transport across the mucosa. Autoradiographic studies showed that specific binding sites for [125I]gastrin releasing peptide were associated with the epithelium of the mucosa as well as the myenteric plexus. Gastrin releasing peptide-like immunoreactive axons paralleled the distribution of specific binding sites for the peptide. These data support a physiological role for gastrin releasing peptide in the control of intestinal ion secretion and motor function.


Subject(s)
Jejunum/chemistry , Peptides , Receptors, Neurotransmitter/analysis , Amino Acid Sequence , Animals , Autoradiography , Biological Assay , Female , Gastrin-Releasing Peptide , Immunohistochemistry , Male , Molecular Sequence Data , Muscle, Smooth/chemistry , Radioligand Assay , Receptors, Bombesin , Sequence Homology, Nucleic Acid , Swine
15.
Peptides ; 11(4): 713-8, 1990.
Article in English | MEDLINE | ID: mdl-1700398

ABSTRACT

The tridecapeptide neurotensin (NT) and its C-terminal homologs, including xenopsin (XP) and neuromedin N (NM-N), reduced the amplitude of spontaneous contractions in longitudinal smooth muscle strips from the porcine distal jejunum in vitro. The rank order of potency (IC50 in nM) was XP (0.1) greater than NT (0.9) approximately avian XP (1.0) greater than NM-N (1.6), which could not be explained on the basis of differential peptide degradation. Tachyphylaxis and cross-tachyphylaxis were observed after repeated NT and XP addition to muscle strips. The action of NT was mimicked by norepinephrine (NE), but not by opioid peptides, somatostatin, or vasoactive intestinal peptide. NE was nearly 100-fold less potent than NT and did not produce a state of tachyphylaxis to NT. The effects of NT and NE were unaltered by the neuronal conduction blocker tetrodotoxin (70 nM). However, the actions of NE, unlike those of NT, were reduced by the alpha-adrenoceptor blocker phentolamine (70 nM), the K(+)-channel blocker apamin (7 nM) and the Ca2(+)-channel blocker verapamil (0.7 microM). These results suggest that NT and related peptides, through a nonadrenergic mechanism, interact with smooth muscle receptors to modulate jejunoileal motor function in the pig.


Subject(s)
Muscle, Smooth/drug effects , Neurotensin/analogs & derivatives , Neurotensin/pharmacology , Animals , Biological Assay , Calcium/pharmacology , In Vitro Techniques , Ion Channels/drug effects , Jejunum/drug effects , Muscle Contraction/drug effects , Norepinephrine/pharmacology , Swine , Tachyphylaxis
16.
Regul Pept ; 29(1): 31-47, 1990 Jun.
Article in English | MEDLINE | ID: mdl-2167504

ABSTRACT

Neuropeptide Y (NPY) and its homolog, peptide YY, are present respectively in neurons and endocrine cells within the mammalian small intestine. In this study, we examined the actions of NPY on ion transport in the porcine distal jejunum mucosa-submucosa in vitro. Peptide YY and NPY were equieffective in producing rapid and sustained decreases in basal short-circuit current (Isc), a bioelectrical measure of active ion transport, eliciting half-maximal decreases at respective serosal concentrations of 0.8 and 30 nmol/l. NPY-induced changes in Isc were due to increased mucosa-to-serosa and net Cl fluxes and were not affected by the absence of extracellular HCO3 ions. NPY activity was correlated with the magnitude of the basal Isc and appeared to depend on the spontaneous production of eicosanoids. The peptide also decreased Isc stimulated by forskolin and 8-bromo-cyclic AMP, but the ionic bases for this effect were complex and differed from those determined under basal conditions. NPY attenuated increases in Isc produced by electrical stimulation of enteric neurons with an IC50 = 5 nmol/l. The actions of the peptide on basal and cyclic AMP-induced ion transport were abolished by the neuronal conduction blocker tetrodotoxin, but not by the opiate antagonist naloxone. The alpha-adrenoceptor blocker phentolamine diminished the effects of NPY on basal, but not cyclic AMP-induced Isc. These results indicate that NPY is capable of modulating NaCl transport in the porcine jejunal mucosa under several different conditions. Furthermore, the effects of the peptide are mediated in part through noradrenergic nerves as well as enteric neurons of unknown chemical identity.


Subject(s)
Cyclic AMP/metabolism , Electrolytes/metabolism , Jejunum/drug effects , Neurons/drug effects , Neuropeptide Y/pharmacology , Animals , Chlorides/metabolism , Colforsin/pharmacology , Electric Stimulation , Female , In Vitro Techniques , Intestinal Mucosa/drug effects , Intestinal Mucosa/innervation , Intestinal Mucosa/metabolism , Jejunum/innervation , Jejunum/metabolism , Male , Membrane Potentials/drug effects , Neurons/physiology , Sodium/metabolism , Swine , Tetrodotoxin/pharmacology
17.
Peptides ; 11(3): 497-500, 1990.
Article in English | MEDLINE | ID: mdl-1696375

ABSTRACT

The enteric neuropeptide galanin (GAL) increased the amplitude of spontaneous contractions in longitudinally oriented muscle strips and inhibited short-circuit current (Isc) elevations induced by transmural electrical stimulation (ES) of mucosal sheets from porcine jejunum in vitro. GAL-induced contractions (GAL EC50 = 9 nmol/l) were maximally 25% of those elicited by 10 mumol/l carbamylcholine and remained unaffected by atropine, tetrodotoxin, or tachyphylaxis to substance P. The presynaptic Ca2+ channel blocker, omega-conotoxin (0.1 mumol/l), inhibited GAL-induced contractions by 66%. GAL attenuated mucosal Isc elevations induced by ES with an IC50 = 13 nmol/l and at 0.1 mumol/l produced rapid decreases in basal Isc averaging 8 +/- 2 microA cm-1 in 77% of tissues examined. The alpha-adrenoceptor blocker phentolamine or the opiate antagonist naloxone did not alter tissue Isc responses to GAL. These results suggest that GAL modulates neuronal activity linked to secretomotor function in the porcine small intestine.


Subject(s)
Intestinal Mucosa/metabolism , Jejunum/physiology , Muscle, Smooth/physiology , Neuropeptides/physiology , Peptides/physiology , Animals , Biological Transport/physiology , Electric Stimulation , Female , Galanin , Ions , Male , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Swine
18.
Regul Pept ; 28(1): 11-22, 1990 Mar 27.
Article in English | MEDLINE | ID: mdl-2158127

ABSTRACT

Canine neurotensin (NT) and neuromedin N (NMN) were isolated from extracts of ileal mucosa using radioimmunoassay for detection. The structures determined were consistent with those predicted by earlier cDNA work. The molar ratio of NT to NMN was ca. 7, suggesting that the NT/NMN precursor, which contains one copy of each peptide, undergoes complex posttranslational processing or that other NT-precursors lacking NMN exist. In addition to NT, small quantities of NT6-13 and NT2-13 were obtained. Native and synthetic preparations of these peptides were indistinguishable in a radioreceptor assay employing rat brain membranes and 125I-labeled NT; NT6-13 was ca. 8-times more potent than NT and NMN was about one-sixth as potent as NT. NT6-13 was also ca. 10 times more potent than NT in inhibiting spontaneous contractile activity in longitudinally-oriented smooth muscle strips of porcine jejunum. Preparations of intestinal N-cells as well as N-cell vesicles also appeared to contain NT2-13 and NT6-13; however, it is not yet clear whether these peptides are utilized physiologically or simply represent metabolites of NT. These results suggest that further work on the processing of NT precursor and on biologic abilities of partial sequences of NT could be fruitful.


Subject(s)
Ileum/analysis , Intestinal Mucosa/analysis , Jejunum/analysis , Neurotensin/analysis , Peptide Fragments/analysis , Amino Acid Sequence , Animals , Dogs , Female , Guinea Pigs , Jejunum/drug effects , Molecular Sequence Data , Muscle Contraction/drug effects , Neurotensin/analogs & derivatives , Neurotensin/isolation & purification , Peptide Fragments/isolation & purification , Pregnancy , Receptors, Neurotensin , Receptors, Neurotransmitter/analysis , Swine
19.
Analyst ; 91(82): 297-305, 1966 May.
Article in English | MEDLINE | ID: mdl-5931546
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