Intact Segments Research Articles

Gender differences in the endothelial regulation of α2-adrenoceptor-mediated contraction in the rat aorta

The aim of this study was to determine the possible influence of sex hormones on the contractile responses induced by clonidine, an agonist of α2-adrenoceptors, as well as the endothelial modulation of these responses. For this purpose, thoracic aorta segments from male (control and castrated) and female (in oestrous phase and ovariectomized) rats were used. In intact segments from the four groups of rats, clonidine (0.01-10 µmol/l) induced concentration-dependent contractions, which were increased by the nitric oxide synthase inhibitor Nω-nitro-⌊-arginine methyl ester (0.1 mmol/l) or by endothelium removal, but were reduced by 1 µmol/l yohimbine (an α2-adrenoceptor antagonist) in all animals and by 1 µmol/l indomethacin (a cyclo-oxygenase inhibitor) in control males only. The rank order of the magnitude of the maximal response was: oestrous females > ovariectomized females > control males > castrated males, whereas the sensitivity to clonidine (EC50 value) was similar in all animals. In endothelium-denuded segments, the rank order was: oestrous females = control males > ovariectomized females = castrated males. These results suggest that: (1) the presence of oestrogen or androgen increases the contraction caused by α2-adrenoceptor activation with clonidine; (2) endothelium negatively modulates the response to this agonist in the four groups of rats, due to endothelial NO release (entirely in females and in part in males); (3) androgen also seems to modulate the response by stimulating the release of an endothelial contracting factor, probably a prostanoid; and (4) the endothelium of males has a greater capacity than that of comparable females for negative regulation of the tension generated by the underlying vascular smooth muscle.

Gender differences in the endothelial regulation of α2-adrenoceptor-mediated contraction in the rat aorta

The aim of this study was to determine the possible influence of sex hormones on the contractile responses induced by clonidine, an agonist of alpha2-adrenoceptors, as well as the endothelial modulation of these responses. For this purpose, thoracic aorta segments from male (control and castrated) and female (in oestrous phase and ovariectomized) rats were used. In intact segments from the four groups of rats, clonidine (0.01-10 micromol/l) induced concentration-dependent contractions, which were increased by the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester (0.1 mmol/l) or by endothelium removal, but were reduced by 1 micromol/l yohimbine (an alpha2-adrenoceptor antagonist) in all animals and by 1 micromol/l indomethacin (a cyclo-oxygenase inhibitor) in control males only. The rank order of the magnitude of the maximal response was: oestrous females>ovariectomized females>control males>castrated males, whereas the sensitivity to clonidine (EC50 value) was similar in all animals. In endothelium-denuded segments, the rank order was: oestrous females=control males>ovariectomized females=castrated males. These results suggest that: (1) the presence of oestrogen or androgen increases the contraction caused by alpha2-adrenoceptor activation with clonidine; (2) endothelium negatively modulates the response to this agonist in the four groups of rats, due to endothelial NO release (entirely in females and in part in males); (3) androgen also seems to modulate the response by stimulating the release of an endothelial contracting factor, probably a prostanoid; and (4) the endothelium of males has a greater capacity than that of comparable females for negative regulation of the tension generated by the underlying vascular smooth muscle.

Auxin Stimulates the Synthesis but not the Loosening of Cell Walls in Isolated Outer Tissue of Dark-Grown Squash ( Cucurbita maxima Duch.) Hypocotyls

Indole-3-acetic acid (IAA) promoted elongation and cell wall loosening of outer tissue of intact segments from squash ( Cucurbita maxima Duch.) hypocotyls. On the other hand, when the segment was separated into the outer and inner tissues and then incubated, the isolated outer tissue slightly shrank and did not elongate, while the isolated inner one rapidly elongated. IAA did not affect either growth or the mechanical properties of cell walls of isolated outer and inner tissues. In intact segments, IAA significantly promoted the incorporation of radioactivity from [ 14 C]glucose into cell wall fractions of outer tissue without affecting uptake into the intact segments but it did not promote that of inner tissue. In isolated outer tissue, IAA substantially inhibited the uptake but significantly promoted the distribution of the incorporated radioactivity into cell wall fractions. IAA, however, barely affected the uptake and incorporation of radioactivity into cell wall fractions in isolated inner tissue. These results suggest that IAA primarily stimulates the synthesis of certain cell wall polysaccharides in outer tissue, irrespective of being intact or isolated, but the stimulation to that extent is not directly involved in the processes of cell wall loosening in outer tissue.

In vitro propagation of Gladiolus hybridus Hort.: Synergistic effect of heat shock and sucrose on morphogenesis

Three cultivars (cvs.) of Gladiolus hybridus Hort., namely ‘Her Majesty’, ‘Aldebaran’ and ‘Bright Eye’ were successfully micropropagated. The cultures were established using intact cormels or segments of cormels and inflorescence axes on Murashige and Skoog (1962; MS) medium. The response depended on media supplements; both callus formation or direct induction of shoot buds was observed. Shoot differentiation from callus could be obtained on MS medium containing 1.0 μM BA (6–benzyladenine) and 10.0 μM NAA (α-naphthalene acetic acid) in all three cultivars. The same could be achieved by giving a heat shock (HS; 50 °C, 1h) to callus cultures (in case of ‘Her Majesty’ and ‘Aldebaran’ only) maintained on the basal medium. In these two cultivars, high sucrose concentration (0.232, 0.290 or 0.348 M) also favoured growth and proliferation of shoot cultures on a plant growth regulator-free medium at 20 °C in comparison to the cultures kept at 25 °C. On the other hand, shoot cultures maintained on the basal medium at 25 °C containing normal (0.058 M, i.e., 2.0%, w/v) sucrose concentration responded similar to those maintained at 20 °C on a high sucrose medium; reduced response was observed on normal sucrose containing medium at 20 °C. Heat shock enhanced shoot proliferation in the cultures maintained on basal medium, but induced prolific rooting in shoot cultures, within 5 days of HS, on high sucrose (optimum 0.232 M) medium. While the number of roots increased at higher sucrose concentrations in the medium in case of cvs. ‘Her Majesty’ and ‘Aldebaran’, the same was found to be independent of sucrose concentration in cv. ‘Bright Eye’. Generally the rooted plants produced on high sucrose (0.232 M) medium in comparison to medium with normal sucrose concentration showed better survival (ca. 90% as against 40%) in the soil.

Contractile responses elicited by hydrogen peroxide in aorta from normotensive and hypertensive rats. Endothelial modulation and mechanism involved.

The present study analyses the influence of hypertension and endothelium on the effect induced by hydrogen peroxide (H2O2) on basal tone in aortic segments from normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) of 6-month-old, as well as the possible mechanisms involved. Single (1 mM) or cumulative (100 nM-10 mM) concentrations of H2O2 produced a transient contraction or a concentration-dependent increase of basal tone, respectively, in segments from WKY and SHR. In both cases, the contractions were higher in intact segments from hypertensive than from normotensive rats, and increased by endothelium removal in both strains. Catalase (1000 u ml(-1), a H2O2 scavenger) abolished the contraction elicited by 1 mM H2O2 in both strains. Superoxide dismutase (SOD, 150 u ml(-1)) and dimethylsulphoxide (DMSO, 7 mM), scavengers of superoxide anions and hydroxyl radicals, respectively, did not alter H2O2-induced contractions in intact segments from both strains. However, L-NG-nitroarginine methyl ester (L-NAME, 100 microM, a nitric oxide synthase inhibitor) increased the response to H2O2 in normotensive rats, although the increase was less than that produced by endothelium removal. Incubation of segments with 1 mM H2O2 for 15 min and subsequent washout reduced the contractile responses induced by 75 mM KCl in intact segments from SHR and in endothelium-denuded segments from both strains; this effect being prevented by catalase (1000 u ml(-1)). Indomethacin (10 microM, a cyclo-oxygenase inhibitor) and SQ 29,548 (10 microM, a prostaglandin H2/thromboxane A2 receptor antagonist) practically abolished the contractions elicited by H2O2 in normotensive and hypertensive rats. We conclude that: (1) the oxidant stress induced by H2O2 produces contractions mediated by generation of a product of the cyclo-oxygenase pathway, prostaglandin H2 or more probably thromboxane A2, in normotensive and hypertensive rats; (2) oxygen-derived free radicals are not involved in the effect of H2O2; (3) in normotensive rats, endothelium protects against H2O2-mediated injury to contractile machinery, determined by the impairment of KCl-induced contractions; and (4) endothelial nitric oxide has a protective role on the contractile effect induced by H2O2, that is lost in hypertension.

Effect of age on the vasorelaxation elicited by cromakalim. Role of K + channels and cyclic GMP

The objective of this study was to investigate whether age induces changes on vasodilator response induced by cromakalim, an ATP-sensitive K + (K ATP) channel opener, as well as the underlying mechanism involved in this possible alteration. For this purpose, aortic segments from young (3–5 months) and old (3 years) rabbits were used, which were precontracted with noradrenaline (NA, 1 μM). The vasodilator response induced by cromakalim (0.01–100 μM) was reduced in intact segments from old rabbits, and endothelium removal reduced and did not modify this effect in young and old animals, respectively. In both groups of animals, glibenclamide (10 μM), a blocker of K ATP channels, significantly reduced the response elicited by cromakalim, which was not modified by the large conductance Ca 2+-activated K + channel blocker charybdotoxin (ChTX, 0.4 μM). Acetylcholine (ACh, 10 μM) and sodium nitroprusside (SNP, 100 μM) induced a lesser vasodilator effect in aortic segments from old compared with young rabbits. In segments precontracted with NA, 10 μM ACh or 100 μM SNP similarly increased cGMP levels in both groups of animals. However, basal cGMP level was reduced in segments from old rabbits. Incubation with 8-bromo-cGMP (100 μM) increased the response induced by cromakalim in both groups of animals, reaching similar maximum values in young and old rabbits. The response induced by cromakalim plus 8-bromo-cGMP was markedly decreased by glibenclamide and unmodified by ChTx in both types of animals. These results suggest that aging decreases the vasodilator response to cromakalim, mechanism in which appears to be involved the maintained low cGMP levels observed in old rabbits, and that this messenger modulates the degree of K ATP channel activation.

Oxidation of NADH by intact segments of soybean hypocotyls and stimulation by 2,4-D

Intact sections of soybean cut from regions of cell elongation of hypocotyls of etiolated soybean seedlings oxidized externally supplied NADH (NADH is an impermeant substrate). The oxidation of NADH by 1-cm intact sections was stimulated by the plant growth factor 2,4-dichlorophenoxyacetic acid (2,4-D). The optimum concentration of 2,4-D for stimulation was about 1 μM. Stimulations also were given by the naturally occurring 2,4-D analog, indole-3-acetic acid (IAA), but not by the growth inactive 2,4-D analog 2,3-dichlorophenoxyacetic acid (2,3-D). The findings confirm studies comparing inside-out and right side-out vesicles that show the 2,4-D-stimulated NADH oxidase to be located at the external cell surface. Since plant cells are unlikely to encounter NADH at their external cell surface, functions of the oxidase in reactions other than oxidation of NADH are discussed.

Fracture behaviour and pullout analysis in ceramic matrix composites using m0dified Rosen's model

A modified Rosen's model is proposed to predict the ultimate tensile load and pullout in ceramic matrix composite materials. The model assumes a Weibull distribution for the fibre strength and that the matrix is already cracked. The solid is discretised in fibre segments between matrix cracks. After fracture of one fibre segment, the load is redistributed among the intact neighbouring segments. This in turn produces further fibre failures and subsequent load transfer. Then, the load is increased on the testpiece until new segments are fractured, and the procedure is iterated until one fracture path isfound. The model is applied to a silicon carbide fibre/calcium aluminium silicate (SiCf/CAS) material and compared with experimental results. It is concluded that upper and lower bounds can be obtained using different load transfer assumptions.

Leafroll Virus Is Common in Cultivated American Grapevines in Western New York.

American grapevines (Vitis labrusca L. 'Niagara'; Vitis × labruscana L. H. Bailey 'Concord' and 'Catawba'; V. labrusca × V. riparia Michx. 'Elvira') from 24 vineyards in the New York portion of the Lake Erie production region (>13,000 ha cultivated) were tested to explore a possible relationship between virus infection and an unexplained fruit set malady in the district. One-year-old cane segments were collected 4 to 6 weeks before budbreak from 65 individual vines, which previously had been identified as malady positive or negative. Preparations from bark scrapings were tested for the presence of double-stranded (ds) RNA and for fan leaf degeneration virus, tobacco streak virus, and grapevine leafroll associated closterovirus-3 (GLRaV-3) by enzyme-linked immunosorbent assay (ELISA). Mechanical transmission of other potential viruses to Chenopodium quinoa was attempted with sap extracted from young shoots forced from intact segments of sampled canes. GLRaV-3 was detected in 17 (26%) of the sampled vines from eight (33%) of the vineyards, but there was no apparent relationship between infected vines and the fruit set malady. Vines of all four cultivars were infected. dsRNA was detected in all 17 samples positive for GLRaV-3 plus four additional samples. No other viruses were detected. Near harvest, nine vines (from two vineyards) previously testing positive for GLRaV-3 were examined and retested; all nine tested positive again, although none showed any overt symptoms of viral infection. This is believed to be the first report of GLRaV-3 from American grape vineyards in New York. The source of these infections is unknown: all vines were self rooted, the individual vineyards had been planted independently at different times, and V. vinifera and its hybrids are rare in the district. Wild grapevines (primarily V. riparia) are abundant in the region, although it has been reported that leafroll disease does not occur naturally in wild North American grapes (1). Nevertheless, our results indicate that cultivated American grapevines can be common reservoirs of GLRaV-3, and furthermore suggest the need to reassess the possibility that wild grapes also may serve as reservoirs of the virus. Trials are currently underway to determine possible effects of GLRaV-3 on cv. Concord, the most widely planted variety in the region. Reference: (1) A. C. Goheen. 1988. Leafroll. Page 52 in: Compendium of Grape Diseases. R. C. Pearson and A. C. Goheen, eds. American Phytopathological Society, St. Paul, MN.

Dopamine inhibits vasopressin-dependent cAMP production in the rat cortical collecting duct.

Dopamine inhibits Na+ and water reabsorption in the rat cortical collecting duct (CCD) in the presence of arginine vasopressin (AVP). This inhibition appears to involve the D4 dopamine receptor isoform, which inhibits cAMP production; however, the D1A receptor, which stimulates cAMP production, is also expressed in the CCD. To discriminate between these opposing effects, we measured cAMP production in intact CCD segments. The basal rate of cAMP production ranged from 6.5 to 10 fmol/mm of tubule length over a 7-min incubation period, and it was unaffected by either dopamine or the D1A-specific agonist fenoldopam. AVP increased cAMP production to the range of 85-153 fmol . mm-1 . 7 min-1. Whereas neither 0.1 nor 1.0 microM fenoldopam affected AVP-dependent cAMP production, dopamine reduced it in a dose-dependent manner, achieving a maximum inhibition of 50% at 10 microM. This effect was reversed by the D4 receptor antagonist clozapine but not by pimozide or spiperone (antagonists of D2 and D3 receptors) or by calphostin C or chelerythrine (inhibitors of protein kinase C). We conclude that dopamine inhibits transepithelial Na+ transport and osmotic water permeability in the presence of AVP by inhibition of cAMP production, which is mediated by the D4 receptor isoform linked via the inhibitory G protein Gi.

Relaxant effects of propofol on human omental arteries and veins

We have investigated the relaxant effects of propofol on smooth muscle tension in human omental arteries and veins to determine if endothelium-related mechanisms are involved. Isolated vessel segments were precontracted with endothelin-1 and propofol was added cumulatively (10(-7)-10(-4) mol litre-1). In both artery and vein segments, propofol induced relaxation, which was not dependent on an intact endothelium. Relaxation was reduced when the extracellular K+ concentration was increased and in the presence of tetraethylammonium chloride (TEA). In intact segments, N-nitro-L-arginine methyl ester (L-NAME), indomethacin, glibenclamide, 4-aminopyridine, clotrimazole and atropine did not affect the concentration-response curve of propofol. This indicates that propofol relaxes human omental arteries and veins in an endothelium independent manner, and that hyperpolarization caused by activation of the K+ channel, BKCa, may be involved.

Association of leaf rust and powdery mildew resistance in a recombinant derived from a Hordeum vulgare × Hordeum bulbosum hybrid

AbstractWhile studying powdery mildew resistance in a recombinant line (code 81882) derived from a Hordeum vulgare (cv. ‘Vada’) ×Hordeum bulbosum hybrid, a low infection type of resistance to leaf rust was observed. To determine the mode of inheritance of the leaf rust resistance and whether there was linkage between the two resistances, F2 and F3 progenies from crosses between 81882 and ‘Vada’ were inoculated with the leaf rust and powdery mildew pathogens. Southern blots were prepared using restricted DNA extracted from leaves of 82 F2 plants and four chromosome 2HS sequences were hybridized with the blots to define the length of the introgression. The leaf rust resistance appears to be inherited as a single dominant gene on chromosome 2HS, which co‐segregates with the powdery mildew resistance. There was an almost complete association between the resistances and the respective molecular markers, but it is likely that the strong linkage results from the frequent inheritance of the introgressed H. bulbosum DNA as an intact segment of chromatin with only low levels of recombination within the segment.

Cl- channels in basolateral renal medullary membranes. XII. Anti-rbClC-Ka antibody blocks MTAL Cl- channels.

Cl- channels in the medullary thick ascending limb (MTAL) studied by either patch-clamp technique or reconstitution into lipid bilayers are activated by increases in intracellular Cl- concentrations. rbClC-Ka, a ClC Cl- channel, may represent this channel. We therefore evaluated the role of rbClC-Ka in transcellular MTAL Cl- transport in two separate ways. First, an antibody was raised against a fusion protein containing a 153-amino acid fragment of rbClC-Ka. Immunostaining of rabbit kidney sections with the antibody was localized to basolateral regions of MTAL and cortical thick ascending limb (CTAL) segments and also to the cytoplasm of intercalated cells in the cortical collecting duct. Second, Cl- uptake and efflux were measured in suspensions of mouse MTAL segments. Cl- uptake was bumetanide sensitive and was stimulated by treatment with a combination of vasopressin + forskolin + dibutyryl adenosine 3',5-cyclic monophosphate (DBcAMP). Cl- efflux was also increased significantly by vasopressin + forskolin + DBcAMP from 114 +/- 20 to 196 +/- 36 nmol.mg protein-1.45 s-1 (P = 0.003). Cl- efflux was inhibited by the Cl- channel blocker diphenylamine-2-carboxylate (154 +/- 26 vs. 70 +/- 21 nmol.mg protein-1.45 s-1, P = 0.003). An anti-rbClC-Ka antibody, which inhibits the activity of MTAL Cl- channels in lipid bilayers, reduced Cl- efflux from intact MTAL segments (154 +/- 28 vs. 53 +/- 14 nmol.mg protein-1.45 s-1, P = 0.02). These results support the view that rbClC-Ka is the basolateral membrane Cl- channel that mediates vasopressin-stimulated net Cl- transport in the MTAL segment.

Wire fixation techniques of the cervical facets.

The changes in the biomechanical responses of the cervical spine altered by multilevel laminectomy to various facet wiring techniques were evaluated. To determine the effectiveness of various proposed techniques of cervical facet wiring used to offer rigid internal fixation after multilevel laminectomy. Eight human cadaveric spine segments from C2-11 underwent combined flexion-compression loading. After testing intact and three-level laminectomy (C4-C6) preparations, two techniques of facet wiring fixation were evaluated in an identical manner. Force, displacement, and kinematics data at every level of the column were obtained. The mean stiffness of the intact column was significantly greater than the mean stiffness for laminectomized specimens. Individual facet wiring to the bone graft and through the spinous process below the laminectomy failed to restore stiffness to the laminectomized preparations, whereas the Luque rectangle method restored the stiffness to that found in the intact column. The increases in segmental and overall sagittal rotations resulting from multilevel laminectomy were not decreased significantly by the individual facet wiring technique, but the Luque rectangle technique demonstrated a reduction of sagittal rotations compared with laminectomy without fixation. The significant increases in total column flexibility and segmental flexural rotations after multilevel laminectomy were not corrected by techniques that depend on individual facet wires secured to an overlying strut, including wiring to the inferior intact segment. Crosslinking of the facet wire fixation above and below the laminectomized segments, as exemplified by the Luque rectangle technique, restored column stiffness and reduced segmental sagittal rotations.

Alpha 1-adrenoceptor subtypes in rat aorta and mesenteric small arteries are preserved during left ventricular dysfunction post-myocardial infarction.

In heart failure, homologous downregulation of beta-adrenoceptors contributes to impaired adrenergic responsiveness of the myocardium. We evaluated alpha 1-adrenoceptors (alpha 1-AR) in a sparsely innervated and a densely innervated peripheral artery in an experimental model of left ventricular dysfunction post-myocardial infarction. [3H]Prazosin binding was determined in arterial segments of Wistar-Kyoto rats (WKY), and of Wistar rats 5 weeks after myocardial infarction (MI) or sham operation (SHAM). In the thoracic aorta (TAO) of WKY, specific prazosin binding was: (i) prevented by the irreversible alpha 1B-AR and relatively selective alpha 1D-AR antagonist, chloroethylclonidine (CEC); (ii) displaced with low affinity (pKi 6.25) by the alpha 1A-AR selective ligand, (+)-niguldipine; and (iii) displaced with both high (pKi 10.4) and low (pKi 7.37) affinity by the alpha 1D-AR antagonist, BMY 7378. In mesenteric small arteries (MSA) of WKY, prazosin binding was: (i) reduced 50% by CEC; (ii) displaced in a biphasic fashion by (+)-niguldipine (pKi 8.60 and pKi 6.22); and (iii) displaced by BMY 7378 with low affinity only (pKi 6.86). Also in TAO of SHAM. prazosin binding was prevented by CEC, but neither 30 nM (+)-niguldipine nor 1 nM BMY 7378 affected it. In MSA of SHAM, prazosin binding was virtually abolished in the presence of 30 nM (+)-niguldipine and was not reduced by 1 nM BMY 7378. In TAO and MSA of MI, compared to SHAM, the density of binding sites tended to be increased rather than decreased and neither the affinity for the ligand nor the effects of alpha 1-AR subtype selective tools were significantly modified. These findings indicate that: (i) radioligand binding can be applied in intact arterial segments to quantify and characterize alpha 1-AR; (ii) although differences seem to exist between rat strains, alpha 1B-AR and alpha 1D-AR predominate in rat thoracic aorta and alpha 1A-AR and alpha 1B-AR in mesenteric small arteries; and (iii) alpha 1-AR density is not reduced in the poorly innervated aorta and the densely innervated mesenteric small arteries of rats with heart failure due to myocardial infarction.

Localization of the spinal network associated with generation of hindlimb locomotion in the neonatal rat and organization of its transverse coupling system.

The segmental organization of the hindlimb locomotor pattern generators and the coordination of rhythmic motor activity were studied in isolated spinal cords of the neonatal rat. All lumbar segments and many thoracic and sacral segments of the cord exhibited an alternating left-right rhythm in the presence of serotonin (5-HT) and N-methyl-D-aspartate (NDMA). Other thoracic segments exhibited a synchronized left-right rhythm or an irregular bursting activity. Transection of the cord at the thoracolumbar or lumbosacral junction abolished the rhythmicity of nonlumbar segments and had no affect on the rhythmicity of lumbar segments. A fast alternating rhythm persisted in rostral lumbar segments after transection of the cord at mid-L3. A much slower alternating rhythm was found in the detached caudal lumbar segments after elevation of the NMDA concentration. These findings suggest that neurogenesis of hindlimb locomotion is not restricted to L1/L2, and that the lumbar pattern generators exhibited rostrocaudal specialization. An alternating left-right rhythm persisted in lumbar cords of midsagittally split preparations that were kept with either L1, L2, L3, or L4 as the only bilaterally intact segment. An alternating rhythm persisted also in preparations that were midsagittally split up to T13-T12, or down to L4. Extension of these lesions led to a bilaterally synchronous rhythm or to left-right independent rhythms in the lumbar cord. These results indicated that the transverse coupling system in the caudal-thoracic and lumbar segments in specialized and that left-right alternation in the lumbar cord can be carried out by the cross connectivity, which is relayed at least through the T12-L4 segments. Bath application of the glycine receptor antagonist strychnine, or the gamma-aminobutyric acid-A (GABAA) receptor blocker bicuculline, induced in the presence of NMDA and 5-HT a bilaterally synchronous rhythm in any intact or detached segment of the cord and in midsagittally split preparations with few bilaterally intact upper thoracic or lower sacral segments. A strychnine-resistant left-right alternating rhythm was found in the presence of 5-HT and NMDA in preparations that were treated with the non-NMDA receptor blocker 6-cyano-7-nitroquinoxaline (CNQX) before and during the application of strychnine. Subsequent washout of CNQX immediately induced a bilateral synchronous rhythm. These results suggest that the phase relation between the hemicords during the rhythm is determined by a dynamic interplay between the excitatory and inhibitory cross connectivity, and that this interplay can be modulated experimentally. Local application of strychnine to L2 kept bilaterally intact in midsagittally split preparations perturbed but did not completely block the alternating pattern of the rhythm induced by 5-HT and NMDA. Local application of bicuculline under the same conditions prolonged the cycle time and had no effect on left-right alternation. These results, together with those described above, suggest that left-right alternation is mediated mainly by strychnine-sensitive glycine receptors with possible contribution of strychnine-resistant glycine receptors and/or GABAA receptors.

Source of ATP, ADP, AMP and adenosine released from isolated rat caudal artery exposed to noradrenaline.

Purines of ATP, ADP, AMP and adenosine released from rat caudal artery with and without endothelium and the isolated smooth muscle and endothelial cells were examined, in order to determine the source. Treatment of intact segments of caudal arteries with noradrenaline (10 microM) for 3 min induced a large release of ATP, ADP, AMP and adenosine. However, if the artery segments had been denuded of their endothelial lining, noradrenraline induced only a slight release of purines. Endothelial cells in primary culture prepared from caudal arteries, when exposed to noradrenaline for 3 min released large amounts of purines, whereas vascular smooth muscle cells prepared similarly and passaged endothelial cells did not release purines upon exposure to noradrenaline. These results indicate that, of smooth muscle and endothelial cells of the vascular wall, only intact endothelial cells react to alpha-adrenoceptor stimulation by releasing adenine nucleotides and adenosine.

Doberman's Ear Tip Plasty

The techniques of nasal-tip correction include preservation of an intact caudal segment of cartilage as a complete strip and vertical interruption of the cartilaginous arch, with or without preservation of the underlying vestibular skin, as an interrupted strip. This technique can be performed with either the nondelivery (transcartilaginous, retrograde) or the delivery approach. The “Doberman's ear” technique has both the advantages of the complete strip technique with the delivery approach and the suture stabilization of the columellar pillar as in the Goldman tip, preserving the underlying vestibular skin as in the “butterfly” (Ponti) operation.

Calcium channel blockers modify jejunal uptake of D-galactose in rabbits.

Calcium channel blockers modify the intestinal uptake of lipids. This study was undertaken to test the hypothesis that two different types of calcium channel blockers influence the uptake of D-galactose, a sugar absorbed by the sodium-dependent glucose transporter (SGLT1) in the intestinal brush border membrane. Nisoldipine (1 mg/kg/day) or verapamil (4 mg/kg/day) were given by mouth to New Zealand white rabbits for three weeks, and then the rates of uptake of varying concentrations (2-64 mM) of galactose were examined in an in vitro preparation of jejunum using the incorporation of 14C-labeled substrate into intact tissue segments. The maximal transport capacities (Vmax) for D-galactose were increased in animals given nisoldipine or verapamil, as compared to controls. The value of the apparent Michaelis constant Km* for D-galactose was higher with nisoldipine group and lower with verapamil, than in controls. The apparent passive permeability (Pd*) of D-galactose was estimated from the uptake of L-glucose: Pd* was lower with nisoldipine and higher with verapamil, as compared to controls. The effect of these drugs on sugar uptake is not due to differences in the animals' food intake, body weight gain, or mucosal surface area. Thus, the two different classes of calcium channel blockers, the dihydropyridine nisoldipine and the phenylalkylamine verapamil, have different effects on the K(m)* and Pd*, but not on the Vmax of D-galactose uptake.

Differential membrane protein phosphorylation in bovine retinal rod outer segment disk membranes as a function of disk age.

The outer segment portion of photoreceptor rod cells is composed of a stacked array of disk membranes. Newly formed disks are found at the base of the rod outer segment (ROS) and are relatively high in membrane cholesterol. Older disks are found at the apical tip of the ROS and are low in membrane cholesterol. Disk membranes were separated based on their membrane cholesterol content and the extent of membrane protein phosphorylation determined. Light induced phosphorylation of ROS disk membrane proteins was investigated using magic angle spinning 31P NMR. When intact rod outer segment preparations were stimulated by light, in the presence of endogenously available kinases, membrane proteins located in disks at the base of the ROS were more heavily phosphorylated than those at the tip. SDS-gel electrophoresis of the phosphorylated disk membranes subpopulations identified a phosphoprotein species with a molecular weight of approximately 68-72 kDa that was more heavily phosphorylated in newly formed disks than in old disks. The identity of this phosphoprotein is presently under investigation. When the phosphorylation reaction was carried out in isolated disk membrane preparations with exogenously added co-factors and kinases, there was no preferential protein phosphorylation. Taken collectively, these results suggest that within the ROS there is a protein phosphorylation gradient that maybe indicative of co-factor or kinase heterogeneity.