Shaping Ability of ProTaper Next, WaveOne, and Reciproc in Simulated Root Canals.

INTRODUCTION: The study aimed to compare the shaping and preservation of the original curvature of simulated curved root canals using the following instruments: Reciproc (Rcp), WaveOne (Wo), and the ProTaper Next system (Ptn). MATERIALS AND METHODS: A total of 45 resin blocks with simulated curved root canals were divided into three groups (n = 15), prepared using the Rcp (R25), Wo (25/0.8), and Ptn (X2) instruments. Standardized photographs were taken before and after canal instrumentation. After the superimposition of the images, the amount of resin removed from the curvature's inner and outer walls was measured at six apical levels, at intervals of 1 mm. The canals' angles of curvature before and after instrumentation were subtracted. RESULTS: There were no significant differences between the instruments in terms of the total amount of resin removed of the inner or outer walls of the apical curvature (p > 0.05). The Rcp instruments provided the best resin removed ratios between the walls. The means of the change in angle were as follows: Wo = 2.15°, Ptn = 0.92°, and Rcp = 0.21°. WaveOne caused significantly higher deviations than Rcp. CONCLUSION: All of the instruments demonstrated a tendency to straighten the simulated root canal. Instruments that use rotary movement achieved an effect similar to that of the reciprocating instruments in relation to change in angle. CLINICAL SIGNIFICANCE: Deviations from the original shape of the root canal could have a negative impact on the quality of a filling and consequently on the success of the endodontic treatment.

Growth patterns of Italian local chicken populations.

Predictions of growth are important factors that contribute to the profitability of an operation in poultry production. Modern commercial hybrids have a higher body growth in comparison with the local purebreds. However a niche market for meat and egg poultry production needs to be established using local purebreds to promote biodiversity. The aim of this study was to model the growth response of male and female chickens belonging to 5 local Italian populations: a commercial slow-growing hybrid (Berlanda, B), the Padovana pure breed [2 plumage varieties: silver, argentata (PA) and chamois, camosciata (PC)], and their crosses PC×B and PC×PA. A total of 398 one-day-old birds were reared until 180 d of age under indoor conditions. The linear and 3 nonlinear models (logistic, Gompertz, and Richards) were compared to study the growth patterns of these chicken populations. Significant (P < 0.01) differences were observed among the genotypes for several curve parameters. In males, PC×B showed the lowest age at inflection point, B showed the highest age and BW, whereas PA showed the highest age and the lowest weight. In females, the age at the inflection point did not differ among the groups; B showed the highest weight. All the nonlinear models gave a good fit of male and female data with R(2) ranging from 0.992 and 0.999, but the logistic equation had higher value of root mean square error than the Gompertz and the Richards values. Based on residual sum of squares for both sexes, the Richards model was better (P < 0.05) than the logistic but not superior to the Gompertz. The logistic equation showed an overestimation of initial BW for all the groups and sex. For Italian local chicken populations, the Richards model requires a measure of BW recorded at 90 d or after to obtain a good fit of the asymptotic weight. However, the Gompertz model has the advantage that it requires one less parameter than the Richards model.

Evaluation of an aerosol emitter for mating disruption of Cydia pomonella in Italy.

Some techniques have been developed to disrupt mating (MD) of codling moth (CM) by treating orchards with pheromone. Synthetic pheromone is applied to the crop as a formulation that is designed to protect these generally labile compounds from degradation while gradually releasing pheromone into the atmosphere. In Trentino South Tyrol MD has been adopted successfully (24,500 ha, i.e. 73% of the apple area) to control CM in heavily infested areas; while in areas with low pest pressure, less pesticides are usually applied (2-3 per year) and as a consequence, pheromone mating disruption is not considered economically convenient. Hand applied sealed plastic tubes and plastic ampoules are the two pheromone formulations more widely used. A new pheromone-based control technique, called Puffer, has been recently proposed. Puffers are battery-powered devices that release pheromone from pressurized aerosol cans every 15 minutes for 12 hours or 30 min for 24 hours. During each puff a quantity of 6.95 mg a.i. is emitted. The high release rate of pheromone per puff from aerosol dispensers is thought to compensate for their low application densities (2-2.5 puffer/hectare). Results of three year field trials carried out in Trentino-South Tyrol demonstrated the potential of Puffer as effective tool to control the moth.

Quality of organic eggs of hybrid and Italian breed hens.

The quality of eggs laid by 2 hybrid and 2 Italian dual-purpose genotypes of hens reared under organic farming system was studied. Hens belonging to Hy-Line Brown (brown eggshell, HLB), Hy-Line White (white eggshell, HLW), Ermellinata di Rovigo (brown eggshell, E), and Robusta maculata (brown eggshell, R) genotypes were reared from 24 to 43 wk of age. The trial was carried out from July to December, with environmental temperature ranging from 25°C (±5°C, summer) to 13°C (±7°C, autumn). The HLB eggs were heavier (P < 0.01) than HLW (62.9 vs. 60.4 g), and R eggs were heavier (P < 0.01) than E (56.5 vs. 54.4 g). The albumen weight differed (P < 0.01) among HLB, HLW, R, and E (40.7, 38.3, 32.7, 34.1 g, respectively). The E and R yolk weights were similar (16.2 g) and higher (P < 0.01) than hybrids; HLW yolk was higher (P < 0.05) than HLB (15.8 vs. 15.5 g). The HLB showed the highest (P < 0.01, 6.74 g) shell weight and E had the lowest (P < 0.01; 5.43 g). The yolk cholesterol content was higher (P < 0.01) in the Italian eggs than in the hybrids (258 vs. 219 mg/yolk). The HLB yolk had the lowest (P < 0.01) saturated fatty acids (33.8 vs. 34.9%), and R yolks showed the lowest (P < 0.01) monounsaturated fatty acids (36.3 vs. 38.0%) and the highest (P < 0.01) polyunsaturated fatty acids (28.7 vs. 27.4%) than the other groups. The HLW yolk showed the highest (P < 0.01) n-6/n-3 ratio (13.7) in comparison to the other 3 groups (12.8). During 21 d of storage (at 21°C and 62% RH), the E eggs showed the lowest (P < 0.01) quality (albumen height, 4.93 vs. 5.56 mm; Haugh units, 71 vs. 74). A sensory profile of boiled eggs showed differences (P < 0.05) in odor and flavor sensations and in certain yolk and albumen texture properties according to genotype. The quality of organic eggs from different genotypes differs in relation to the strain but also the interaction with the environmental conditions has to be considered.

Chemical composition of meat and egg yolk of hybrid and Italian breed hens reared using an organic production system.

A trial was done to study the chemical composition of meat and egg yolk of laying hens belonging to different genotypes reared using an organic production system. Two commercial hybrid hens (Hy-Line Brown and Hy-Line White 36) and 2 Italian dual-purpose breed hens (Ermellinata di Rovigo and Robusta Maculata) were reared from 25 to 44 wk of age. During the experimental period (July to December), the environmental temperature decreased from about 25 to 13 degrees C and the RH increased from 62 to 73%. The photoperiod was 16L:8D. Meat and egg yolks of 44-wk-old hens were analyzed. The Italian breeds showed higher meat production and lower egg production when compared with the hybrids. Genotype affected protein, lipid, and ash content of breast and thigh meat. Hy-Line White and Ermellinata di Rovigo showed the lowest (P<0.01) breast lipid and Hy-Line White had the lowest (P<0.01) thigh lipid. Hy-Line White showed higher (P<0.05) breast and thigh cholesterol. Genotype affected egg yolk cholesterol, which was higher (P<0.01) in Italian breeds than in hybrids. The fatty acid profiles of meat and yolk were significantly affected by genotype. The hybrid and Robusta Maculata meat and egg fatty acid composition may be considered to be more favorable to human health. The results indicate that the ability of laying hens to incorporate fatty acids is genetically dependent and a possible interaction between strain and environmental conditions may exist for the absorption and utilization of dietary components.

Environmental risk assessment of the anionic surfactant sodium lauryl ether sulphate in site-specific conditions arising from mechanized tunnelling.

Sodium lauryl ether sulphate (SLES) is the anionic surfactant commonly utilized as the main synthetic chemical component in most foaming agents used in mechanized tunnelling. This produces huge amounts of soil debris which can contain residual concentrations of SLES. The absence of environmental quality standards for soil and water and the limited information about SLES persistence in real excavated soils do not facilitate any re-use of soil debris as by-products. The environmental risk assessment (ERA) of foaming agents containing SLES can be a valid tool for this purpose. In this study, an ERA analysis of SLES in 12 commercial formulations (cf) used for tunnelling excavation was performed. Various soils from different tunnel excavation sites were conditioned with the selected foaming agents containing SLES. Predicted or measured environmental concentrations (PECs, MECs) were determined and then compared with the Predicted No Effect Concentrations (PNECs) for both the terrestrial and aquatic compartments. The results indicate a reduction of the potential risk over time for these ecosystems, with differences depending on both the commercial foaming formulation and the spoil material characteristics. However, because potential threats to the natural environment cannot be excluded, some risk management and mitigation actions are discussed.

Spatial-temporal analysis and risk characterisation of pesticides in Alpine glacial streams.

We analysed the spatial and temporal distribution of a selection of pesticides in Alpine glaciers used on the Po Plain in Northern Italy, near the Alps. By analysing a 102-m ice core taken from the Lys Glacier (Monte Rosa massif, Italy), we highlight historical contamination from the insecticide chlorpyrifos and the herbicide terbuthylazine, confirming the role of alpine glaciers as temporal sinks. In addition, we collected meltwater samples from six glaciers distributed along the Alpine Arc during the summer of 2016, which showed widespread contamination by pesticides. Overall, chlorpyrifos and terbuthylazine dominated the contaminant fingerprint of all of the studied glaciers, with contamination peaks occurring at the beginning of the melting season. This highlights the importance of the medium-range atmospheric transport of these pesticides in connection with agricultural practices in the areas beneath the Italian Alps, where they are widely applied. The release of pesticides in meltwater can lead to potential risks to the aquatic ecosystems of headwater streams, as we demonstrate for chlorpyrifos. This suggests that the medium-range atmospheric transport of pesticides should be considered as part of regulations to protect the water quality of these pristine environments.

Transport Properties of Li-TFSI Water-in-Salt Electrolytes.

Water-in-salts are a new family of electrolytes that may allow the development of aqueous Li-ion batteries. They have a structure that is reminiscent of ionic liquids, and they are characterized by a high concentration of ionic species. In this work, we study their transport properties and how they evolve with concentration by using molecular dynamic simulations. We first focus on the choice of the force field. By comparing the simulated viscosities and self-diffusion coefficients with experimental measurements, we select a set of parameters that reproduces well the transport properties. We then use the selected force field to study in detail the variations of the self and collective diffusivities of all the species as well as the transport number of the lithium ion. We show that correlations between ions and water play an important role over the whole concentration range. In the water-in-salt regime, the anions form a percolating network that reduces the cation-anion correlations and leads to rather large values for the transport number compared to other standard electrolytes.

Effect of genotype on slaughtering performance and meat physical and sensory characteristics of organic laying hens.

Slaughtering yields and some meat physical and sensorial parameters of laying hens reared under organic system production were studied. The hens belonged to both Italian dual-purpose breeds [Ermellinata di Rovigo (ER; brown eggshell) and Robusta Maculata (RM; brown eggshell)] as well as hybrid genotypes [Hy Line White 36 (white eggshell) and Hy Line Brown (brown eggshell)]. The birds were reared under organic farming system production from 24 to 44 wk of age, when they were slaughtered. They were reared throughout summer and autumn, and the temperature ranged from about 28 to 3 degrees C. Local breeds presented higher (P < 0.01) live BW and dressing percentage compared with hybrids. The RM and ER carcasses had (P < 0.01) the highest breast and leg (thigh and drumstick) percentage, respectively. The muscle-bone ratio of the Hy Line White 36 drumstick was lower (P < 0.05) than the RM ratio, whereas the other groups were intermediate. The ER breast presented the highest (P < 0.01) lightness value and the lowest (P < 0.01) final pH value compared with the other 3 groups. The breast meat significantly differed according to genotype for almost all the studied sensorial parameters (adhesivity, fibrousness, chewiness, solubility, juiciness, tenderness, shear resistance), with the exception of aroma and odor intensity. In the thigh, genotype significantly affected aroma, adhesivity, fibrousness, solubility, tenderness, and shear resistance.

Home and office uroflowmetry for evaluation of LUTS from benign prostatic enlargement.

A group of 107 patients with lower urinary tract symptoms (LUTS) from benign prostatic enlargement (BPE) participated to the HOUSE Study (Home and Office Uroflowmetry Specific Evaluation). Patients received routine investigation, consisting of medical history taking, physical examination including digital rectal examination, prostate-specific antigen (PSA), assessment of symptoms listed both on the International Prostate Symptom Score and on ICS-male questionnaire. We examined the results of uroflowmetry evaluation in this population; data were analysed to observe if any circadian changes of parameters obtained with home uroflowmetry could be detected. We searched a correlation between Q(max), Q(ave) and ICS-benign prostatic hyperplasia symptom score: a significantly inverse correlation was found only for Q(max), confirming Q(max) as a reliable parameter to quantify subjective symptoms. When examining the multiple flow curves recorded in the same patient with home uroflowmetry, voided volume and flow time had usually higher values during night-time: the existence of circadian changes of uroflowmetry parameters in patients with LUTS from BPE was confirmed, and lower values of average and maximum flow rates during sleep hours were recorded in the same patient. In conclusion, when evaluating the natural history or treatment outcome of individual patients or group of patients in clinical trials for evaluation of BPE and LUTS, an assessment including multiple measurements may be useful and of value.

Exercise induces myonuclear ubiquitination and apoptosis in dystrophin-deficient muscle of mice.

Apoptosis plays a major role in several diseases, including viral infections, autoimmune diseases, cancer, cardiac infarct, and neurological disorders. To investigate the role of apoptosis in muscular dystrophy, dystrophin-deficient (mdx) mice were subjected to spontaneous exercise and skeletal muscles were analyzed for apoptosis and ubiquitin. The increase of apoptotic myonuclei after exercise was detected by TUNEL, by electron microscopy, and by DNA analyses for high molecular weight and for ladder fragments. Expression of ubiquitin correlated with exercise and with positive myonuclei for apoptosis. Biochemical analysis confirmed a high level of ubiquitination both in sarcoplasmic and myofibrillar proteins. Muscles from sedentary congenit control mice (C57B ) were negative for apoptosis, while after exercise some nuclei were positive. We also revealed that normal myoblasts committed to apoptosis in vitro showed an increased expression of ubiquitin. Western blot for bcl-2, FasL, and BAG1 showed a significant decrease of bcl-2 product only in mdx mice after exercise. Thus, spontaneous exercise results in the increase of ubiquitin expression and in the reduction of bcl-2 tightly related to programmed cell death in mdx mice. These findings confirm that DNA fragmentation, absent in muscles of sedentary normal mice but present in mdx mice at rest, dramatically increases after exercise, shedding new light on exercise-induced muscle damage and on its progression in dystrophinopathies.

Apoptosis, DNA damage and ubiquitin expression in normal and mdx muscle fibers after exercise.

The current view indicates that after eccentric exercise myofibers are mechanically damaged and therefore an inflammatory and necrotic process occurs. In the present paper we examine the possibility that apoptosis plays a role in normal and dystrophin-deficient muscles after running. We analysed for apoptosis normal and dystrophin-deficient mouse muscles after a night of spontaneous wheel-running followed by two days of rest. Terminal deoxynucleotidyl transferase-mediated end-labeling of DNA in nuclei in tissue sections and gel electrophoresis of extracted DNA showed the presence of fragmented DNA. Furthermore, ubiquitin, a protein whose appearance is related to apoptosis, increased in muscles of both dystrophic and normal runner mice. The present findings which confirm that DNA damage is absent in muscles of sedentary mice but present in muscles of runner mice offer a new hypothesis on early events of muscle damage.

Regulation of plasma aldosterone levels by metoclopramide: a reappraisal of its mechanism from dopaminergic antagonism to serotonergic agonism.

It has been thought, since the late 1970s, that dopamine exerts a tonic suppression of plasma aldosterone levels in human subjects. This action, however, had not been established directly using dopamine and dopamine mimetic drugs, which do not, in fact, affect the aldosterone levels. Rather, the conclusion was arrived at indirectly, based on the increase in aldosterone levels seen with dopamine receptor blockers; metoclopramide in particular, considered at the time of its discovery in the 1960s to be a new generation dopamine antagonist. However, metoclopramide is not a pure drug and in fact, shows intermediate affinity at certain serotonin receptor subtypes. Studies have been recently carried out in human subjects on the role of serotonergic transmission in mediating the metoclopramide as an aldosterone secretagogue effect. Here we briefly review this work and attempt to reassess the action of metoclopramide as an aldosterone secretagogue, from dopamine D2 antagonism to serotonin 5-HT4 partial agonism.

Identification of serotonin 5-HT4 recognition sites in the porcine caudate nucleus by radioligand binding.

Specific binding for the serotonin 5-HT4 receptor (5-HT4R) radioligand [3H]GR 113808 was identified in pig caudate nucleus and characterized by serotonin subtype selective drugs. Binding was inhibited by serotonin and by synthetic indoles, benzamides and benzimidazolones known to characterize the 5-HT4R in functional tests. Rank order of potency of 5-HT4R antagonists was: GR 125487 (Ki, 0.19 nM) > GR 113808 >> SC 53606 > SDZ 205,557 > RS 235971/190 > DAU 6285 > tropisetron > DAU 6215. GR 125487 and GR 113808 were highly selective with respect to the 5-HT3 receptor (5-HT3R). Rank order of potency of 5-HT4R agonists was: SC 53116 (Ki, 21 nM) > BIMU 1 > cisapride > BIMU 8 > serotonin > renzapride > S-zacopride > metoclopramide > R-zacopride > 5-methoxytryptamine >> 5-carboxamidotryptamine. BIMU 8, renzapride, metoclopramide and the zacopride enantiomers gave shallow competition curves. The agonists were substantially less selective than the antagonists with respect to the 5-HT3R. With only two exceptions, SCH 23390 and metergoline, which bound with sub-microM affinity to the 5-HT4R, binding was not inhibited by compounds selective for other G-protein-coupled or channel-gated receptors. Highly significant correlations in affinities of compounds for 5-HT4R in caudata of pigs, guinea pigs and humans were found suggesting no difference among mammalian species.

The role of GABAA receptor function in peristaltic activity of the guinea-pig ileum: a comparative study with bicuculline, SR 95531 and picrotoxinin.

1. The peristaltic activity of the guinea-pig ileum was studied in the absence and in the presence of the blockade of GABAA receptors. 2. Bicuculline (1-30 microM), improved at the highest concentrations the efficiency of peristalsis by enhancing the frequency of propulsive contractions and the amount of fluid ejected per unit of time. 3. Neither SR 95531 (0.3-10 microM), a novel GABAA receptor antagonist, which competitively antagonized 3-aminopropane sulphonic acid induced contractions in myenteric plexus-longitudinal muscle preparations (pA2 value: 6.47), nor picrotoxinin (1-30 microM) modified peristaltic parameters or influenced the potentiating effect of bicuculline on peristaltic activity. 4. In myenteric plexus-longitudinal muscle preparations, bicuculline (1-30 microM) enhanced the amplitude of electrically-induced cholinergic contractions without modifying submaximal contractions to applied acetylcholine. SR 95531 and picrotoxinin had no effect on twitch amplitude. In the presence of each of these compounds, bicuculline retained its potentiating effect. 5. The results obtained with SR 95531 and picrotoxinin question the view that GABAA receptors may exert a critical role in intestinal propulsion by modulating the activity of nerve pathways subserving peristalsis. Bicuculline potentiates the peristaltic activity of the ileum probably via a facilitatory effect on enteric cholinergic transmission that is independent of GABAA receptor blockade.

5-Hydroxytryptamine receptors that facilitate excitatory neuromuscular transmission in the guinea-pig isolated detrusor muscle.

1. In isolated detrusor strips from the guinea-pig urinary bladder, contractile responses to electrical field stimulation were mostly mediated by neurally released acetylcholine (ACh) and adenosine 5'-triphosphate (ATP). 2. 5-Hydroxytryptamine (5-HT) produced a concentration-dependent increase in the amplitude of stimulated detrusor strip contractions. The 5-HT concentration-response curve showed a biphasic profile: the high potency phase was obtained at sub-micromolar concentrations (10-300 nM), while the low potency phase in the range 1-30 microM. The maximum response of the first phase was 30% of the total 5-HT response. 3. Like 5-HT, the 5-HT3 receptor agonist, 2-methyl-5-hydroxytryptamine (2-methyl-5-HT: 0.3-100 microM), the 5-HT2 receptor agonist, (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI: 30 nM-3 microM) and the 5-HT4 receptor agonist, 5-methoxytryptamine (5-MeOT: 0.1-30 microM) potentiated, though with lower potency, detrusor contractions. The resulting concentration-response curves were monophasic in nature. 2-Methyl-5-HT had a maximum effect comparable to that of 5-HT. By contrast, the maximal effects of DOI and 5-MeOT were only 20% and 30% of that elicited by 30 microM 5-HT, respectively. 4. The 5-HT3 receptor antagonist, granisetron (0.3 microM) had no effect on the high potency phase, but caused a rightward parallel shift of the low potency phase of the 5-HT curve (pKB = 7.3). Granisetron(0.3 microM) antagonized with comparable affinity (pKB = 7.1) 5-HT-induced responses after pharmacological isolation of 5-HT3 receptors with the 5-HT1/5-HT2 receptor antagonist, methiothepin (0.3 microM) and the 5-HT4 receptor antagonist, GR 125487 (30 nM). Granisetron (0.1, 0.3 and 1 microM) competitively antagonized the potentiating effect of 2-methyl-5-HT with an estimated pA2 of 7.3.5. Methiothepin (0.3 microM) and the 5-HT2A receptor antagonist, ketanserin (0.3 microM) produced a slight inhibition of the first phase of the 5-HT curve. In the presence of ketanserin, an equimolar concentration of methiothepin was ineffective in further reducing the effect of 5-HT. Similarly, the 5-HT4 receptor antagonist, GR 125487 (30 nM) slightly inhibited the first phase of the 5-HT curve. Conversely, this phase was suppressed when detrusor strips were coincubated with ketanserin (or methiothepin) and GR125487.6. In a separate set of experiments, the interactions of 5-HT with either the purinergic or cholinergic components of excitatory neuromuscular transmission were investigated. In the presence of hyoscine(1 microM), 5-HT was mostly effective at sub-micromolar concentrations, while in the presence of the P2-purinoceptor antagonist, suramin (300 microM), 5-HT-induced potentiation was mainly obtained with micromolar concentrations.7. Thus, in electrically stimulated detrusor strips from guinea-pig, 5-HT potentiated excitatory neuromuscular transmission by activating at least three separate neural 5-HT receptors. These include the 5-HT2A and 5-HT4 receptors, which mediate the 5-HT high potency phase mainly by activation of purinergic transmission. On the other hand, the potentiating effect caused by micromolar concentrations of 5-HT mostly involves cholinergic transmission and is mediated by the 5-HT3 receptors.

The influence of neuronal 5-hydroxytryptamine receptor antagonists on non-cholinergic ganglionic transmission in the guinea-pig enteric excitatory reflex.

A partitioned bath made it possible to separate the site of recording of the ascending excitatory reflex of the ileal circular muscle (oral compartment) from the site of reflex induction (caudal compartment), evoked by inflating an intraluminal balloon. In the caudal compartment, blockade of cholinergic ganglionic transmission by hexamethonium (100 microM) and hyoscine (0.3 microM) caused an approximately 65% reduction in the amplitude of reflex contractions, suggesting that the remaining response was mediated by non-cholinergic transmission near the distension site. This non-cholinergic component of ganglionic transmission was insensitive to the action of methiothepin (1 microM), ondansetron (1 microM), tropisetron (1.5 microM), DAU 6285 (1 microM) and renzapride (1 microM), agents that antagonize the action of 5-hydroxytryptamine (5-HT) at neural 5-HT1-like, 5-HT3, 5-HT4 and putative 5-HT1P receptors. These findings suggest that the neural pathways subserving non-cholinergic ganglionic transmission in the ascending excitatory reflex in the guinea-pig ileum do not involve 5-HT as neurotransmitter.

An in vitro study of the relationship between GABA receptor function and propulsive motility in the distal colon of the rabbit.

1. The effects of gamma-aminobutyric acid (GABA), 3-aminopropane sulphonic acid (3-APS) and baclofen on spontaneous, electrically-induced and propulsive motility were investigated in rabbit distal colon. 2. In unstimulated longitudinal (LMPs) and circular muscle strip preparations (CMPs) 3-APS (10-200 microM) and GABA caused a clear-cut relaxation susceptible to desensitization. Baclofen (10-200 microM) caused relaxation in a minority (30%) of preparations. The 3-APS response was sensitive to tetrodotoxin (TTX; 1 microM), SR 95531 (a novel competitive GABAA-receptor antagonist) (10 microM), picrotoxinin (30 microM), and insensitive to hyoscine (1 microM) and to a combination of prazosin (1 microM) and propranolol (1 microM). The baclofen response was antagonized by 5-aminovaleric acid (DAVA, 500 microM), TTX and hyoscine and resistant to GABAA-receptor and adrenoceptor blockade. GABAA-receptors were therefore associated with non-adrenergic non-cholinergic (NANC) inhibitory nerve activation while GABAB-receptors were involved in depression of cholinergic tone of smooth muscle. GABA (10-200 microM) elicited both above mentioned effects. 3. In LMPs, baclofen (10-200 microM) dose-dependently inhibited submaximal responses to both cholinergic and NANC inhibitory nerve stimulation. This effect was resistant to SR 95531 and picrotoxinin and prevented by DAVA and baclofen desensitization. GABA (10-200 microM) mimicked the action of baclofen. GABA inhibitory effects persisted in the presence of GABAA-receptor blockade. 4. In segments of distal colon, GABA and baclofen (1-200 microM), but not 3-APS (1-200 microM), dose-dependently decreased the velocity of propulsion of an intraluminally-distended balloon. This effect was antagonized by DAVA and GABA or baclofen desensitization and resistant to SR 95531 and picrotoxinin. These antagonists per se had no effect on propulsion. In preparations in which propulsion was slowed by hyoscine (1 microM), baclofen caused no consistent further depression of propulsive activity. 5. Our results show that GABAA- and GABAB-receptors are present in rabbit colon. GABAA-receptor stimulation activates NANC inhibitory nerves without apparently affecting propulsion. GABAB-receptors are associated with a reduction of neural (mainly cholinergic) activity subserving muscular tone and peristalsis and appear to be located on both cholinergic and NANC inhibitory nerves. However, the persisting propulsive activity during suppression of GABAA- and GABAB-receptor function suggests that GABA in enteric neurones is not crucial for the neural circuitry subserving colonic peristalsis in this species.

Characterization of the 5-HT receptor potentiating neuromuscular cholinergic transmission in strips of human isolated detrusor muscle.

In human isolated detrusor strips, submaximal contractile responses evoked by electrical stimulation were resistant to hexamethonium (30 microM) and abolished by tetrodotoxin (0.6 microM) and hyoscine (1 microM), indicating the activation of postganglionic cholinergic nerves. In methysergide (1 microM) and ondansetron (3 microM) pretreated tissues, 5-hydroxytryptamine (5-HT) (0.3 nM-1 microM) caused a concentration-dependent increase in the amplitude of contractions (pEC50 = 8.1), which was antagonized by the selective 5-HT4 receptor antagonist GR 113808 (3, 10 and 30 nM) in a competitive manner. Schild analysis yielded a pA2 estimate of 8.9, a value comparable to that reported for GR 113808 in other animal and human peripheral tissues (8.8-9.7). Our findings indicate that neuromuscular cholinergic transmission in human isolated detrusor muscle is facilitated by neural 5-HT receptors belonging to the 5-HT4 subtype. The human urinary bladder can thus be regarded as an additional site in which 5-HT4 receptors are distributed.

Inhibitory action of capsaicin on cholinergic responses induced by GABAA agonists in the guinea-pig ileum.

Pretreatment of the guinea-pig ileum with capsaicin resulted consistently in depression of the neurogenic cholinergic contractions induced by the GABAA receptor agonists 3-aminopropane sulphonic acid (3-APS) and muscimol. Since capsaicin acts mainly by releasing and depleting substance P from its stores in intestinal nerves, it is likely that substance P plays a role in the response caused by GABAA-mimetic compounds, On the whole, our results suggest that excitatory responses to 3-APS and muscimol result from both direct and indirect activation of intrinsic intestinal cholinergic neurons innervating smooth muscle cells.