A five-year retrospective study on ascarid infections in dogs in southern Italy.

INTRODUCTION: A 5-year retrospective analysis of ascarid infections (Toxocara canis and Toxascaris leonina) in dogs from southern Italy was performed to update the epidemiological scenario of these parasites and to identify the risk factors which may favour these infections in animals in this study area. A total of 8,149 dogs, referred to our labs for copromicroscopic analysis using the FLOTAC technique, was considered. A sub-sample of 500 faecal samples were analysed also with the Mini-FLOTAC technique. Of the overall dog samples analysed, 9,2 % (95 % CI = 8,6-9,8) resulted positive for T. canis while 0,5 % (95 % CI = 0,4-0,7) resulted positive for T. leonina. Co-infections with T. canis and T. leonina were found in 0,1 % of dogs (95 % CI = 0,0-0,1). The results obtained by the FLOTAC and Mini-FLOTAC examinations showed a nearly perfect k agreement (k = 0,99, P < 0,001) between these two techniques. Chi-square test showed positivity to T. canis and T. leonina significantly (P < 0,001) associated with dogs housed outdoor (i.e., that lived in garden or in kennel). Moreover, the positivity for T. canis was significantly associated (P < 0,001) also with age (i.e., puppies), as shown by the logistic regression. The decreasing overall prevalence both for T. canis and T. leonina during the years of monitoring, showed that, as suggested by the European Scientific Counsel Companion Animal Parasites, the regular diagnosis could contribute to an efficient control of these parasites.

INTRODUCTION: Une analyse rétrospective sur 5 ans des infections à ascaris (Toxocara canis et Toxascaris leonina) chez les chiens du sud de l'Italie a été réalisée afin de mettre à jour le scénario épidémiologique de ces parasites et d'identifier les facteurs de risque pouvant favoriser ces infections chez les animaux de cette zone d'étude. Au total, 8149 chiens ont été analysés dans notre laboratoire avec une analyse copromicroscopique en utilisant la technique FLOTAC. De plus, un sous-échantillon de 500 échantillons fécaux a été analysé avec la technique Mini-FLOTAC. Sur l'ensemble des échantillons fécaux canins analysés, 9,2 % (IC à 95 % = 8,6 à 9,8) se sont révélés positifs pour T. canis tandis que 0,5 % (IC à 95 % = 0,4 à 0,7) ont été positifs pour T. leonina. Des co-infections avec T. canis et T. leonina ont été trouvées chez 0,1 % des chiens (IC à 95 % = 0,0­0,1). Les résultats obtenus par les examens FLOTAC et Mini-FLOTAC ont montré un coefficient Kappa presque parfait (k = 0,99, p < 0,001) entre ces deux techniques. Le test du chi carré a montré une positivité significative quant aux infections à T. canis et T. leonina (P < 0,001) associées à des chiens hébergés à l'extérieur (jardin ou chenil). De plus, la positivité pour T. canis était également significativement associée (P < 0,001) à l'âge (c'est-à-dire aux chiots), comme le montre la régression logistique. La diminution de la prévalence globale au cours de la période de surveillance a montré que le diagnostic régulier pourrait contribuer à un contrôle efficace de ces parasites à la fois pour T. canis et T. leonina, comme suggéré par le the European Scientific Counsel Companion Animal Parasites.

Bradykinin-induced contractions of canine saphenous veins: mediation by B2 receptors and involvement of eicosanoids.

1. Experiments were designed to determine the subtype of kinin-receptors mediating the contraction of venous smooth muscle to bradykinin and to investigate the involvement of metabolites of arachidonic acid in this response. 2. Bradykinin (10(-9) to 10(-6) M) caused concentration-dependent contractions of the canine isolated saphenous vein without endothelium, which were potentiated by indomethacin (10(-5) M, an inhibitor of cyclo-oxygenase). The concentration-response curve was biphasic, reaching an asymptote at 10(-8) M and a secondary maximal response at 10(-6) M. 3. Bradykinin (10(-8) M to 3 x 10(-6) M) caused a three fold stimulation in the release of the vasodilator prostaglandin E2 (PGE2) and a two fold stimulation of that of the vasodilator prostacyclin, measured by the production of 6-keto-PGF1 alpha (its stable breakdown product). 4. Under control conditions, nordihydroguaiaretic acid (NDGA, 10(-5) M), an inhibitor of lipoxygenase, did not affect the response to bradykinin. In the presence of indomethacin (10(-5) M), NDGA reduced contractions to bradykinin, suggesting the involvement of lipoxygenase metabolites in the potentiation evoked by the inhibitor of cyclo-oxygenase. 5. The selective B1 receptor agonist [des-Arg9]-bradykinin, in the concentration-range 10(-6) to 10(-5) M, induced contractions, which were abolished by the B2 receptor antagonist D-Arg-[Hyp3, Thi5, D-Tic7, Oic8]-bradykinin (Hoe 140; 10(-6) M). The selective B1 receptor antagonist [des-Arg9, Leu8]-bradykinin, (10(-7) to 10(-5) M) had no significant effect on bradykinin-induced contractions. 6. The B2 receptor antagonists Hoe 140 (10(-8) to 10(-6) M) and D-Arg [Hyp3, D-Phe7]-bradykinin (10(-7) to 10(-5) M) shifted the concentration-response curve to bradykinin to the right in a concentration-dependent manner. 7. These results indicate that, in the canine saphenous vein, bradykinin causes contraction by activating B2 receptors. This results in the production of metabolites of arachidonic acid, which play a key role in the contraction of canine saphenous venous smooth muscle.

Rilmenidine activates postjunctional alpha 1- and alpha 2-adrenoceptors in the canine saphenous vein.

Experiments were performed to determine the subtypes of alpha-adrenoceptors involved in the contraction induced by rilmenidine in isolated canine cutaneous veins. Rings of saphenous vein (without endothelium) were suspended for the recording of isometric force in physiological salt solution. All experiments were performed in the presence of propranolol (to antagonize beta-adrenoceptors), cocaine (to inhibit neuronal uptake) and hydrocortisone (to inhibit extraneuronal uptake). In the presence of rauwolscine (an alpha 2-adrenergic blocker), rilmenidine caused concentration-dependent contractions which were inhibited by prazosin (nonselective alpha 1-antagonist) and by (+)niguldipine (selective alpha 1A-adrenergic antagonist), but not by (-)niguldipine. After treatment with phenoxybenzamine (to alkylate alpha 1-adrenoceptors), rilmenidine evoked contractions of the canine saphenous vein which were antagonized competitively by rauwolscine. The combination of rauwolscine and prazosin did not abolish contractions evoked by the highest concentrations of rilmenidine. Although binding experiments using 3H-idazoxan suggested the existence of a nonadrenergic binding site (around 20% of the total binding), contractile studies failed to demonstrate their involvement in the increases in tension evoked by rilmenidine. These experiments suggest that the contractions evoked by rilmenidine in isolated canine veins are mediated by both alpha 1A- and alpha 2-adrenoceptors.

Regulation of nitric oxide-like activity by prostanoids in smooth muscle of the canine saphenous vein.

1. Organ bath experiments and measurements of prostanoids were performed to investigate the presence of nitric oxide synthase in venous smooth muscle and its interaction with cyclo-oxygenase. 2. In rings of canine saphenous vein without endothelium, the inhibitor of cyclo-oxygenase, indomethacin (10 microM), induced contraction. NG-nitro-L-arginine (100 microM) (L-NOARG), an inhibitor of nitric oxide synthase did not affect the tone of rings of canine saphenous vein when administered alone. However, in the presence of indomethacin L-NOARG (100 microM) induced further contraction. 3. Similar results were obtained in response to NG-monomethyl-L-arginine (L-NMMA)(300 microM or NG-nitro-L-arginine methylester (L-NAME)(100 microM). 4. When rings of canine saphenous vein without endothelium were contracted with phenylephrine (1 microM) instead of indomethacin, neither L-NOARG or L-NMMA induced further contraction. 5. When rings of canine saphenous vein without endothelium were contracted with noradrenaline (0.3 microM) in the presence of indomethacin (10 microM) plus L-NOARG (100 microM), a relaxation to L-arginine was observed. Transient relaxations to superoxide dismutase (150 u ml-1) were observed in all rings. 6. When rings of saphenous vein without endothelium were incubated with lipopolysaccharide (LPS) (100 micrograms ml-1) or interleukin-1 beta (10 u ml-1) the concentration-contraction curve to noradrenaline was not affected. 7. Rings without endothelium released prostaglandin E2 and prostaglandin I2, as measured by radioimmunoassay. The basal production was abolished by indomethacin and not affected by L-NOARG. 8. These results suggest that when cyclo-oxygenase is inhibited, a nitric oxide synthase activity is revealed in rings of canine saphenous vein without endothelium.

cGMP mediates the desensitization to bradykinin in isolated canine coronary arteries.

The relaxation to bradykinin in canine coronary arteries is mediated by endothelium-derived nitric oxide (NO) and hyperpolarizing factor (EDHF). Desensitization to the kinin was induced by incubation of canine coronary arteries with endothelium with 10(-8) M bradykinin for 30 min. After washout, tissues were contracted with prostaglandin F2 alpha, and concentration-relaxation curves to bradykinin were obtained in control and desensitized arteries treated with indomethacin. After desensitization, there was a shift to the right of the concentration-relaxation curves to bradykinin. However, the elevation in guanosine 3',5'-cyclic monophosphate (cGMP) levels evoked by bradykinin was similar in both groups of tissues. The curves to bradykinin obtained in the presence of NG-nitro-L-arginine (an NO synthase inhibitor) were depressed, whereas those obtained in arteries contracted with potassium (to eliminate the EDHF-mediated relaxation) were not affected by the desensitization. Addition of NG-nitro-L-arginine, oxyhemoglobin, or methylene blue before the desensitization procedure preserved, whereas 3-morpholinosydnonimine (SIN-1, a donor of NO) and 8-bromoguanosine 3',5'-cyclic monophosphate impaired, the EDHF-mediated relaxation to bradykinin. Thus the selective impairment of the EDHF-dependent relaxation to bradykinin may be mediated by NO, acting mainly through increased production of cGMP.

Potentiation by trandolaprilat of the endothelium-dependent hyperpolarization induced by bradykinin.

In canine coronary arteries, bradykinin evokes endothelium-dependent relaxations that are mediated by nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF). The converting-enzyme inhibitor trandolaprilat potentiates the endothelium-dependent relaxations evoked by bradykinin in this tissue. The present experiments were designed to determine whether or not facilitated release of EDHF contributes to the augmented response to bradykinin in the presence of trandolaprilat. Organ-chamber studies were performed to measure changes in isometric tension in rings of canine coronary arteries. In the presence of nitro-L-arginine, an inhibitor of NO synthase, trandolaprilat augmented the endothelium-dependent relaxations evoked by bradykinin. These relaxations were not inhibited by the K(+)-channel inhibitors tetraethylammonium, 4-aminopyridine, or glibenclamide, but were abolished in high-potassium solution. The membrane potential in individual smooth-muscle cells of coronary artery was measured by means of glass microelectrodes. Trandolaprilat potentiated the endothelium-dependent hyperpolarizations evoked by a subthreshold concentration of bradykinin, and these endothelium-dependent hyperpolarizations were inhibited by high-potassium solution. These experiments demonstrate that EDHF contributes to the relaxation evoked by bradykinin in the canine coronary artery and that trandolaprilat potentiates the release of this factor. This effect of trandolaprilat may contribute to its vasodilator properties.

Endothelium-dependent effects of converting-enzyme inhibitors.

Angiotensin-converting enzyme (ACE) inhibitors were designed to prevent the vasoconstrictor influence of the activated renin-angiotensin system. However, it has long been suspected that the vasodilator actions of these compounds are not entirely related to inhibition of the generation of angiotensin II. Bradykinin, which is rapidly degraded by ACE, stimulates the release of endothelium-derived vasodilator mediators, including nitric oxide, endothelium-derived hyperpolarizing factor, and prostacyclin. These mediators do not contribute to the vasodilator effect of bradykinin in every arterial bed. However, the prevention by ACE inhibitors of the degradation of bradykinin-induces an augmentation of the production of these substances and thus potentiates the dilatation evoked by the peptide. The existence of a local kallikrein-kinin system in the vascular wall has been demonstrated, and locally generated kinins contribute to the acute vasodilator actions of ACE inhibitors. ACE inhibitors can potentiate endothelium-dependent dilatations evoked by neurohumoral mediators that are not substrates for ACE. Thus, the vasodilator properties of ACE inhibitors not only reflect inhibition of the renin-angiotensin system but also depend on the enhanced production of endothelium-derived mediators.

The sydnonimine C87-3754 evokes endothelium-independent relaxations and prevents endothelium-dependent contractions in blood vessels of the dog.

Experiments were designed to compare the relaxing activities of the new sydnonimine C87-3754 with SIN-1 in arteries and veins of the dog, and to determine whether C87-3754 can prevent endothelium-dependent contractions. Rings of coronary and femoral arteries, and saphenous veins were suspended in organ chambers for the measurement of changes in isometric tension. SIN-1 and C87-3754 evoked concentration-dependent relaxations in all rings of blood vessels contracted with a submaximal concentration of either prostaglandin F2 alpha, endothelin-1, phenylephrine, or norepinephrine. In both arteries and veins, the concentration-relaxation curves to C87-3754 were shifted significantly to the right (by two to three logarithmic units) of that to SIN-1. The presence of endothelium significantly inhibited the relaxations to SIN-1 but did not affect those to C87-3754. The treatment of coronary arteries with methylene blue or oxyhemoglobin significantly impaired the relaxation to SIN-1 and C87-3754. Neither C87-3754 nor its prodrug pirsidomine (CAS 936) affected the membrane potential in coronary arteries. The endothelium-dependent contractions evoked by nitro L-arginine, arachidonic acid, and the calcium ionophore A23187 in basilar arteries of the dog were inhibited by C87-3754. These results indicate that the sydnonimine C87-3754 is a dilator of both arterial and venous smooth muscle, and can prevent endothelium-mediated contractions in cerebral arteries of the dog. The inhibition of vascular tone is likely to involve the activation of soluble guanylate cyclase, causing enhanced production of cyclic guanosine monophosphate in the smooth muscle without a change in membrane potential.

Calmidazolium, a calmodulin inhibitor, inhibits endothelium-dependent relaxations resistant to nitro-L-arginine in the canine coronary artery.

1. The role of calmodulin in endothelium-dependent relaxations in the canine coronary artery, was investigated by use of the inhibitor of calmodulin, calmidazolium. 2. The endothelium-dependent relaxations to adenosine diphosphate (ADP) and nebivolol, a beta-adrenoceptor antagonist, in control solution, and to bradykinin in high potassium solution (to inhibit endothelium-dependent hyperpolarization), were abolished by nitro-L-arginine (30 microM), an inhibitor of nitro oxide-synthase. Calmidazolium (10 microM) did not inhibit these relaxations. 3. Calmidazolium did not affect the endothelium-independent relaxations to SIN-1, an exogenous donor of nitric oxide (NO). 4. The relaxations to bradykinin and to the calcium ionophore A23187 in control solution were inhibited to a small extent by calmidazolium (10 microM). 5. Bradykinin and A23187 induced relaxations in the presence of nitro-L-arginine (30 microM) that were abolished by calmidazolium (10 microM) but not affected by glibenclamide (10 microM), an inhibitor of ATP-sensitive K+ channels. 6. The endothelium-independent relaxations to lemakalim, an ATP-sensitive K+ channel opener, were not affected by calmidazolium (10 microM) but were inhibited by glibenclamide (10 microM). 7. These results suggest that calmidazolium does not inhibit the endothelium-dependent relaxations due to endothelium-derived NO in the canine coronary artery but inhibits either the production of endothelium-derived hyperpolarizing factor (EDHF) from endothelial cells or its effects on vascular smooth muscle cells. Furthermore these results suggest that EDHF contributes to endothelium-dependent relaxations in the canine coronary artery.

Heterogeneous distribution of endothelium-dependent relaxations resistant to NG-nitro-L-arginine in rats.

Endothelium-dependent relaxations that are resistant to inhibitors of nitric oxide synthase probably are mediated by endothelium-dependent hyperpolarization of the vascular smooth muscle. Experiments were performed to examine the distribution of this type of relaxation along the arterial tree of the rat by measuring changes in isometric force. Acetylcholine induced concentration- and endothelium-dependent relaxations in aortas and in pulmonary, common iliac, femoral, mesenteric, and renal arteries contracted with phenylephrine. In the presence of NG-nitro-L-arginine, the cumulative administration of acetylcholine induced relaxations only in the femoral, mesenteric, and renal arteries. The calcium ionophore A23187 relaxed mesenteric arteries contracted with phenylephrine in a concentration- and endothelium-dependent manner. The concentration-relaxation curve to A23187 was shifted to the right in the presence of NG-nitro-L-arginine. The maximal relaxations induced by lemakalim, a K+ channel opener, were smaller in those arteries that did not exhibit NG-nitro-L-arginine-resistant relaxations. These results suggest that NG-nitro-L-arginine-resistant relaxations are more frequently observed in smaller arteries. The arteries that exhibit NG-nitro-L-arginine-resistant relaxations may be more sensitive to an endothelium-derived substance that causes hyperpolarization of vascular smooth muscle cells.

Calmodulin antagonists inhibit endothelium-dependent hyperpolarization in the canine coronary artery.

1. The effects of the calmodulin antagonists, calmidazolium and fendiline were investigated on endothelium-dependent hyperpolarization in the canine coronary artery. The membrane potential of vascular smooth muscle cells was measured with the microelectrode technique. 2. Smooth muscle cells of the canine coronary artery had a resting membrane potential of -50 mV. Bradykinin and the Ca(2+)-ionophore, A23187, induced concentration- and endothelium-dependent hyperpolarization. The hyperpolarization induced by a supramaximal concentration of bradykinin (10(-6) M) reached approximately 20 mV. 3. Calmidazolium (10(-5) M) and fendiline (10(-4) M) inhibited hyperpolarization induced by bradykinin and A23187. By contrast, calmidazolium did not affect the hyperpolarization induced by lemakalim, an opener of ATP-sensitive K(+)-channels. 4. These observations suggest that calmodulin is involved in the generation of endothelium-dependent membrane hyperpolarization of vascular smooth muscle.

Potentiation of endothelium-dependent relaxations to bradykinin by angiotensin I converting enzyme inhibitors in canine coronary artery involves both endothelium-derived relaxing and hyperpolarizing factors.

Studies were designed to investigate the mechanisms underlying the augmentation by angiotensin I converting enzyme (ACE) inhibitors of the endothelium-dependent relaxations evoked by bradykinin. Isometric tension, tissue levels of cGMP, and transmembrane potential were measured in isolated canine coronary arteries as indications of the respective contribution of nitric oxide and endothelium-derived hyperpolarizing factor. In rings of coronary artery with endothelium, relaxations to bradykinin were potentiated by the ACE inhibitors cilazaprilat and perindoprilat. NG-Nitro-L-arginine (NLA), a nitric oxide synthase inhibitor, impaired relaxations to bradykinin. But the presence of ACE inhibitors partially restored this activity. Bradykinin stimulated the production of cGMP, and this was enhanced significantly by ACE inhibitors, indicating an augmented release of nitric oxide. NLA abolished the increase induced by bradykinin irrespective of the presence of ACE inhibitors. Electrophysiological studies revealed that bradykinin elicited an endothelium-dependent hyperpolarization of vascular smooth muscle that was insensitive to NLA and potentiated by ACE inhibitors. The bradykinin-induced hyperpolarization and NLA-resistant relaxations were transient and impaired by potassium depolarization. Thus, production of endothelium-derived hyperpolarizing factor may account for the NLA-resistant relaxations of canine coronary arteries. The relaxations induced by bradykinin were unaffected by the B1 kinin receptor antagonist des-Arg9,[Leu8]-bradykinin either in the absence or in the presence of NLA but were antagonized by the B2 kinin receptor antagonist D-Arg[Hyp3,D-Phe7]-bradykinin. Molecular exclusion chromatography of 125I-labeled [Tyr8]-bradykinin and its degradation products demonstrated that the breakdown of the kinin by isolated coronary arteries was prevented in the presence of perindoprilat.(ABSTRACT TRUNCATED AT 250 WORDS)

Local production of kinins contributes to the endothelium dependent relaxations evoked by converting enzyme inhibitors in isolated arteries.

The angiotensin converting enzyme (ACE) inhibitor perindoprilat evokes endothelium-dependent relaxations in perfused isolated canine arteries. Kininogens, the precursors of bradykinin, elicit endothelium-dependent relaxations which are potentiated by perindoprilat, inhibited by B2-kinin antagonists and partially impaired after inhibition of NO synthase. These observations suggest that locally produced kinins may stimulate the production of NO and endothelium-derived hyperpolarizing factor, and that this action is potentiated by ACE inhibitors.

Decrease in enkephalinase A number in kidney membranes from hypercholesterolemic and hypertensive rats.

The variation of enkephalinase A number on the hypertensive and hypercholesterolemia rats kidney membranes is studied using the [3H]-acetorphan, a potent inhibitor of enkephalinase A to label the protease in rat kidney. The binding of [3H]-acetorphan to kidney membrane determined in vitro with both equilibrium and kinetic methods is saturable and reversible involving a single class of sites with a dissociation constant of 4-5.3 nM. The [3H]-acetorphan binding capacity is identical, Bmax approximately 51 pmoles per mg of proteins, for kidney membranes from Sprague Dawley and Wistar Kyoto rats. In contrast, the enkephalinase A number is decreased in the pathological states studied: 20% for hypertensive rats and 50% for hypercholesterolemic rats. Such pharmacological results provide a great deal of information about the modification appeared in the metabolism of peptidic substrates of enkephalinase A in hypercholesterolemia and hypertension.

Indications for early surgery in cases of lumbar and dorsolumbar scoliosis: the role of vertebral rotation.

There are very few objective criteria for the choice between conservative and surgical treatment of lumbar and dorsolumbar scoliosis ranging from 40 to 50 degrees. We reviewed the long-term results obtained in 76 patients treated with plaster braces because they had refused surgical treatment; in 56% of the cases a nearly 10 degree gain was maintained after the onset of treatment; all of the cases which showed improvement presented a reduction in the curve by at least half and rotation by at least one-third in tests in suspension or in bending, and this was maintained in plaster. In cases where the long-term results were poor, however, rotation did not change. In conclusion, of the many parameters examined, only a combined assessment of the reducibility of both the Cobb angle and of rotation provides a valid indication for the treatment of lumbar and dorsolumbar curves ranging from 40 to 50 degrees.

[Correlations between the prick test and aspecific bronchial reactivity in atopic patients]. / Correlazioni tra prick-test e reattività bronchiale aspecifica in pazienti atopici.

Aspecific bronchial provocation test with Methacholine plays an outstanding role in the diagnostic routine of asthmatic disease, in order to accomplish a correct interpretation of the relation between A.B.I. and bronchial asthma. All researches give a special prominence to the bronchial reactivity distribution among the population, and point out that the distribution curve of response to aspecific challenge shows itself as an unimodal curve, since there is no clinic group (neither that one including subjects defined asthmatic from a clinic or anamnestic point of view) which clearly separates from the rest. A further important consideration indicates that a positive response to the test is not specific of bronchial asthmatic subjects, but it can also occur in patients affected with other pathologies such as chronic bronchitis, allergic rhinitis, and so on. Our study aimed, therefore, to evaluate the correlations occurring between skin sensitivity and aspecific bronchial reactivity in 5 groups of atopic patients (affected with: (1) asthma; (2) asthma + rhinitis; (3) asthma + rhinitis + conjunctivitis; (4) rhinitis + conjunctivitis; (5) rhinitis). In doing such correlations we also considered some other factors like sex, smoking habit, town or country provenance. Sensitiveness, specificity, and predicting value of Prick-test in comparison with Methacholine test have been analysed as well. The results so obtained show that no correlation occurs between Prick-test and Aspecific Bronchial Test in the groups of tested subjects. Test sensitiveness increases in the groups of patients affected with associated pathologies, and depends on factors like sex, smoking habit, town or country provenance.