Autonomic neuropathy in BB rats and alterations in bladder function.

In vivo urinary bladder function was examined in BB rats after 4 and 6 mo of diabetes, and the data were correlated with morphometric changes in the pelvic and hypogastric nerves, which constitute the micturition reflex arc. After controlled bladder distension, diabetic animals revealed irregular bladder contractions at frequencies that were reduced to 33% of normal values and with significantly increased amplitudes. The abnormal micturition in diabetic animals was elicited at moderately elevated threshold volumes. These functional abnormalities of the diabetic bladder were associated with a progressive axonopathy of afferent myelinated sensory fibers and later-occurring axonal atrophy of unmyelinated efferent preganglionic fibers. These data suggest that diabetic urinary bladder dysfunction is initiated by a visceral sensory neuropathy involving the afferent limb of the micturition reflex arc.

Load dependence of isovolumic relaxation in intact heart: facts or artifacts?

In order to study the dependence of left ventricular isovolumic relaxation on preload, afterload, and contractility the effects of infusions of dextran, phenylephrine, and dobutamine were assessed in 10 closed chest anaesthetised dogs. Left ventricular and aortic pressures and left ventricular transverse diameters were measured by micromanometers and a tracking sonomicrometer. Isovolumic relaxation time constant was computed by two different single exponential models: the first (time constant Tw) assumed the horizontal asymptote as equal to zero, whereas the second (time constant Tl) assumed a variable asymptote (Pb). To compare the two models, deviations between observed and predicted left ventricular pressures during isovolumic relaxation were computed for both (average squared difference ARSSQw and ARSSQl respectively). Dextran infusion, although increasing preload indexes, did not affect Tl (from 35.1(2.6) to 38.5(2.2) ms, NS) (mean(SEM], but increased Tw (from 28.4(1.4) to 43.8(2.1) ms, p less than 0.001); Pb was significantly shifted upwards (from -7.9(2.4) to +8.2(2.8) mmHg, p less than 0.01). Pb correlated with left ventricular end diastolic pressure (r = 0.71, p less than 0.001). Phenylephrine infusion did not change the isovolumic relaxation time course (Tl from 36.4(3.5) to 46.2(6.1) ms, NS; Tw from 26.8(2.3) to 30.5(2.9) ms, NS) nor Pb (from -9.5(2.3) to -18.7(2.3) mmHg, NS). Dobutamine infusion reduced Tl significantly (from 35.2(3.7) to 25.3(2) ms, p less than 0.02), but did not change Tw (from 27.5(2.4) to 23.3(3.3) ms, NS) nor Pb (from -7.3(1.8) to -8.8(2.3) mmHg, NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of cholesterol-supplemented diet in heritable hyperlipidemic Yoshida rats: functional and morphological characterization of thoracic aorta.

In this study we have considered the possibility of inducing vascular damage in Yoshida Pittsburg (YOS) rat, an inbred strain which has endogenous hyperlipidemia without vascular atherosclerotic damage. Cholesterol-enriched diet (4% cholesterol plus 1% cholic acid and 0.5% thiouracil) was administered to YOS rats, in order to induce atherogenesis. The results indicate that, despite significant increase in serum (about 2-fold) and aortic tissue cholesterol (about 6-fold), no morphological damage occurred. A reduction in acetylcholine-mediated relaxation (of about 37%) was observed. No inhibition of ATP- or sodium nitrite-induced relaxation, or of contraction induced by norepinephrine was seen. Serum triglyceride concentration did not vary after administration of a cholesterol-enriched diet. Our results suggest that in heritable hyperlipidemic Yoshida rat, after 2 months of cholesterol-enriched diet, despite increased serum cholesterol levels, no atheromatous plaque developed on the aortic wall. Impaired vascular function and reductions in the response to acetylcholine were related to changed endothelial cell function. Administration of a high cholesterol diet to YOS rat may represent a new model of mixed endogenous and exogenous hyperlipidemia that can resemble many human dislipidemic diseases and therefore may become a useful tool for the study of isolated endothelial dysfunction.

Acute haemodynamic effects of ibopamine and dopamine on isovolumic relaxation.

The effects of intraduodenal ibopamine (a new orally active inotropic agent claimed to have haemodynamic effects similar to dopamine) on isovolumic relaxation were monitored for 90 min in eight closed-chest anaesthetized dogs. Dopamine and epinine (ibopamine active metabolite) were also infused at graded doses. After 15 min, ibopamine (12 mg/kg) shortened the time constant of isovolumic relaxation, and increased stroke volume and mean aortic pressure. Peak positive dP/dt increased significantly only 10 min later. Heart rate did not change. Dopamine (10 micrograms/kg per min) similarly reduced the time constant, and increased stroke volume, mean aortic pressure, peak positive dP/dt and heart rate. Epinine (10 micrograms/kg per min) caused similar changes in peak positive dP/dt, stroke volume, mean aortic pressure, and accelerated time constant without raising the heart rate. Ibopamine and epinine therefore significantly improved the isovolumic relaxation phase, like dopamine, without however affecting the heart rate.

Inner ester derivatives of gangliosides protect autonomic nerves of alloxan-diabetic rats against Na+, K(+)-ATPase activity defects.

Bovine brain gangliosides have been shown to prevent decay in Na+,K(+)-ATPase activity in sciatic and optic nerves of alloxan- and streptozotocin-diabetic rats. In the search for a drug with greater bioavailability and increased incorporation into neural tissue, ganglioside inner ester derivatives (AGF1) were recently developed. We evaluated the effect of AGF1 treatment on Na+,K(+)-ATPase activity in homogenates of vagus nerve from alloxan-diabetic rats (100 mg/kg s.c.). Animals were treated with AGF1: 10 mg/kg 6 days/week i.p., or 30 mg/kg biweekly i.p. Treatment began 10 d post-alloxan and continued for 8 consecutive weeks. Normal age- and sex-matched rats were used as controls. Alloxan intoxication produced a 39% decrease in Na+,K(+)-ATPase activity of the vagus nerve, which was completely restored (96-97% recovery) by both AGF1 regimes. Results suggest that ganglioside inner ester derivatives may be used in the clinical setting for the management of diabetic autonomic neuropathy.

Time course of pressure and flow in ascending aorta during ejection.

To analyze aortic flow and pressure relationships, 10 closed-chest anaesthetised dogs were instrumented with electromagnetic aortic flow probes and micromanometers in the left ventricle and ascending aorta. Left ventricular ejection time was divided into: time to peak flow (T1) (both pressure and flow rising), peak flow to peak pressure time (T2) (pressure rising, flow decreasing), and peak pressure to dicrotic notch time (T3) (pressure and flow both decreasing). These time intervals were expressed as percent of total ejection time. Load-active interventions rose markedly T2 (from 4.2 +/- 5.5 to 19.4 +/- 3.5 after phenylephrine (p less than 0.02); from 4.2 +/- 6.5 to 21.2 +/- 5.3 after dextran (p less than 0.02)). Conversely, dobutamine reduced T2 from 4.4 +/- 5.9 to -2.5 +/- 6.5 (p less than 0.05). Thus, during load-active interventions aortic pressure increases for a longer T2 time although forward flow is decreasing, as a result of higher aortic elastic recoil during ejection. Conversely, beta 1-adrenergic stimulation significantly shortens T2. Dynamic pressure-flow relationship is thus continuously changing during ejection. T2 seems to be inversely related to the efficiency of left ventricular ejection dynamics.

Cryodamage of the vessel wall accelerates the development of atherosclerotic lesions in arterial vessels of Watanabe hyperlipidemic rabbits.

In the present study we developed an experimental model, resembling human atherosclerosis, by removing the endothelial layer in Watanabe heritable hyperlipidemic (WHHL) rabbits (10 months old) by application of cryodamage on the external surface of arterial vessels. In age-matched New Zealand white (NZW) rabbits, used as control, after two months following cryodamage, carotid artery and infrarenal segments of abdominal aorta did not show any particular change in the ultrastructure of vessel wall. In WHHL rabbits, two months after cryodamage, atherosclerotic lesions (fatty streaks and fibrous plaques) were observed in both arteries. Many lipid-laden endothelial cells, subendothelial foam cells and smooth muscle cells were found in cryodamaged areas. In some areas, the cap of plaques appeared to be thinned and ruptured. Increased number of collagen and elastic fibrils was also observed in atherosclerotic regions. We conclude that this simple technique on WHHL rabbits provides a model of atherosclerosis with a high degree of morphological similarity between the artificially-induced plaque and human atherosclerotic plaque.

Diabetic autonomic neuropathy and bladder function: pharmacological use of gangliosides and insulin.

This paper briefly describes experimental evidence indicating that bladder dysfunction observed in diabetic rats is due in part to hyperdiuresis and in part to autonomic nerves alterations. The latter, in analogy with somatic nerves alteration, can be ameliorated by treatment of diabetic animals with gangliosides. The use of this agent in the autonomic nerve dysfunction of diabetic origin is discussed.

Alterations of the vegetative nervous system, experimental diabetes and pharmacological use of gangliosides.

We studied in diabetic animals several variables related to the vegetative innervation of the heart and the urinary bladder. Cardiac content of norepinephrine was lower in the heart of the diabetic mice (db/db) as compared to the heart of their heterozygote littermates (db/m). Bladder responses were clearly altered in alloxan treated rats as compared to normal rats, as shown by an increased vesical weight, an increased threshold for micturition reflex and a reduced rate of rhythmic contraction. Ganglioside administration was able to prevent partly or completely these alterations observed in the diabetic animals.

Urine retention due to intra-spinal cord injection of colchicine in rats: improved recovery of bladder function by monosialoganglioside GM1 and nerve growth factor administration.

Intra-spinal cord injection of a low dose of colchicine (2 micrograms/rat) at the lumbar level affects the micturition reflex leading to voiding suppression, bladder hypertrophy and overflow incontinence which lasts about four weeks. The administration of nerve growth factor and monosialoganglioside GM1 normalizes urine output within 3 days and improves recovery of the bladder contraction tested by a cystometric analysis.

[Agronomic, nutritional and processing characteristics of soybean germ plasm with colored seed coats]. / Características agronómicas nutricionais e de processamento de germoplasma de soja com tegumentos coloridos.

Ten soybean varieties with colored seed coats were evaluated in Jaboticabal, with the objective of obtaining information as to color preference in the direct use of soybeans in human consumption. The above-mentioned material showed good adaptation to the local environmental conditions: plant cycles were smaller than the Santa Rosa (121 days), varying from 107 to 119 days, in spite of the fact that some varieties started blooming later than Santa Rosa. All materials are within the minimum standards for local planting; however, some of them showed a lodging problem, and all varieties are susceptible to bacterial pustule. They have smaller seeds than Santa Rosa, and in relation to yield, varieties as the NC-55, Aksarben 1S (Black), Aksarben 1S (Brown) and Chi kei 13 did not statistically differ from the Santa Rosa. Protein content showed a variability of 37.90 to 43.90% and oil varied from 14.72 to 21.34%. Methionine content was between the known limits (0.907 to 1.644 g/16 g N), but lysine was higher than any reported data (7.584, to 10.877 g/16g N). The Tanner, Chi kei 13 and Chi kei 15 presented a high percentage of hard beans. This fact had a positive influence on the seed hydration characteristics, but their experimental cooking times were very low, varying from 51 and-a-half to 122 minutes. The term "hydration time" is being introduced here, which is defined as the time, in hours, required for a seed to double up its weight when submerged in water.

Experimental diabetes in the rat: alterations in the vesical function.

Vesical function has been studied in vivo in alloxan-diabetic rats. Rhythmic contractions evoked as a micturition reflex by a controlled distension of the bladder were recorded in urethane-anesthetized animals. These rhythmic contractions are essentially induced by the parasympathetic and inhibited by the sympathetic outflow. The study has been performed on male Sprague-Dawley rats. They were treated at the age of two months with vehicle (control group) or alloxan (100 mg/kg s.c.). Such treatment caused hyperglycemia (greater than 250 mg/dl) and glycosuria in most of the animals. In control animals, analyzed between 5 and 8 months of age, the contractions evoked by vesical distension appeared at a threshold volume between 0.5 and 1 ml, and they had a regular rate. In a first series of experiments, diabetic animals 3 months after treatment showed the evoked response at an average threshold volume of about 1.7 ml. while in animals with 6 months of experimental diabetes the average threshold volume was about 3.2 ml. The rate of contractions in diabetic animals became more and more irregular as the duration of diabetes increased. These marked functional alterations were associated with the evidence of bladder enlargement. In a second series of experiments, bovine brain gangliosides (10 mg/kg i.p./day) or saline were given to diabetic rats between the 30th and the 90th day after alloxan. Ganglioside administration significantly reduced bladder enlargement and the threshold volume for micturition reflex and improved the rate of bladder contractions. It is suggested that alterations of the autonomic nervous system can be expressed as functional vesical dysfunction in experimental diabetes.

[A family with progressive myoclonus epilepsy (author's transl)]. / Eine Familie mit progressiver Myoklonus-Epilepsie.

To begin, a survey of the literature concerning the group of progressive myoclonic epilepsies is presented, from the initial descriptions of Unverricht (1891) and Lundborg (1903) to the present. Recently several subforms of this nosologic entity have been delineated according to the mode of inheritance, time of manifestation, severity of course, and biochemical characteristics (i.e, eventual demonstration of mucopolysaccharide storage in Lafora bodies or diffuse in the central nervous system and other organs). The most useful classification stems from Diebold (1972): early (I) and late (II) forms of the Lafora type having autosomal recessive inheritance; an autosomal recessive early form (III) and an autosomal dominant late form (IV) with degenerative changes in the central nervous system without biochemical disturbances. The authors describe 3 young siblings from Southern Tyrol, who clinically manifested the cardinal symptoms of the disease in addition to extrapyramidal cerebellar disturbances. In spite of extensive bioptic and biochemical examinations, neither Lafora bodies nor diffuse deposits or excretion of mucopolysaccharides could be demonstrated. The distant blood relationship between the parents of these patients supports the assumption of an autosomal recessive mode of transmission. The relatively early manifestation of the disease and the demonstration of degenerative changes within the central nervous system suggest assignment of these patients to Diebold's subgroup III of the progressive myoclonic epilepsy.

[Is it possible to predict the number of nebulizations and the use of intravenous corticosteroid in children with asthma attacks in the emergency room?] / E possível prever o número de nebulizações e o uso de corticosteróide intravenoso em crianças com crise asmática na Unidade de Emergência?

OBJECTIVE: To identify the number of nebulizations needed and the demand for intravenous corticosteroids in children with asthmatic attack, considering clinical and functional characteristics presented at the moment children were admitted to the emergency room. METHODS: We prospectively evaluated 130 children with asthmatic attack and from 1 to 13 years of age. At the moment children were admitted, they were evaluated according to clinical score, arterial oxygen saturation (pulse oximetry), and peak expiratory flow. Next, children received a standard treatment and were observed for the number of consecutive nebulizations required and the use of corticosteroid. Using regression analysis, we assessed the parameters evaluated for a correlation with the number of nebulizations and the use of intravenous corticosteroid. RESULTS: Eighty-eight children (67.7%) received from 1 to 3 nebulizations and 42 children (32.3%), 6 nebulizations. Sixty-eight children (52.3%) received corticosteroid. The initial values for clinical scores, arterial oxygen saturation, and peak expiratory flow showed a significant correlation with the number of nebulizations required and the need for corticosteroids. CONCLUSIONS: According to our results, it is possible to predict and anticipate, at the time chidren are admitted with asthmatic attack, the need for corticosteroid and more nebulizations, which can change the prognosis and the time of evolution of the attack.

Changes in muscle esterases in genetically dystrophic and control littermate mice.

The hypothesis that the existence of a family of isoenzymes is at the basis of the heterogeneity of esterase activity among skeletal muscles from dystrophic Re 129J and control littermate mice was tested using isoelectrofocusing (IEF) in polyacrylamide gel. All muscles considered showed not only quantitative heterogeneity, as previously observed in this laboratory, but also qualitative differences with regard to their esterase isoenzymes. Differences in the number and the relative amounts of isoenzymatic bands were found between different muscles from both control and dystrophic animals. In most dystrophic muscles a new isoenzymatic band appeared at pH 6.6. Another band, identified at pH 5.12, was more pronounced in dystrophic than in control muscles. The specificity of these observations is indirectly supported by the fact that other tissue, such as heart, liver, and kidney, did not show the quantitative or qualitative abnormality present in the dystrophic skeletal muscles.

Power spectrum analysis of cardiovascular variability monitored by telemetry in conscious unrestrained rats.

Beat-to-beat variability of arterial pressure and heart period (R-R) was studied in eight conscious freely-moving adult male rats in which telemetric recordings of arterial pressure, ECG and respiratory movements were obtained under unrestrained and unstressed conditions. The beat-to-beat time series of these signals (systolic arterial pressure, diastolic arterial pressure and R-R) were analyzed, in the frequency domain, using autoregressive spectral analysis in order to detect and quantify the rhythmic components. In basal conditions, the systolic arterial pressure variability spectrum was characterized by three major spectral components which had central frequencies respectively of 0.08 +/- 0.03 Hz (very low frequency), 0.43 +/- 0.02 Hz (low frequency) and 1.36 +/- 0.19 Hz (high frequency). Similar rhythmic components were found in R-R signal variability. The very low frequency component included a higher percentage of total power in R-R variability spectrum (75.3%) than in systolic arterial pressure variability spectrum (58.4%). The low frequency component was more pronounced in both systolic and diastolic arterial pressure variability spectra. The high frequency component of R-R, systolic and diastolic arterial pressure was synchronous with respiration. Cross-spectral analysis revealed a high statistical coherence between R-R and arterial pressure variabilities in all the three frequency bands. An alpha-adrenergic blocker (phentolamine) specifically abolished the low frequency components of systolic and diastolic arterial pressure variability spectra, thus suggesting that low frequency is a marker of sympathetic modulation of vasomotor activity. The low frequency component of R-R variability spectrum was also markedly blunted. We suggest that cardiovascular variability signals, (R-R, systolic and diastolic arterial pressure) are composed almost of two main rhythms linked to respiration and vasomotor activity. These rhythms can be quantified in conscious unrestrained rats by using telemetry and spectral analysis. This approach seems to offer a new powerful tool for pharmacological studies in conscious small animals.

Ibopamine, an orally active dopamine-like drug: metabolism and pharmacokinetics in dogs.

Ibopamine (SB-7505), the 3,4-diisobutyryl ester of N-methyldopamine (epinine), exerts, on oral administration, cardiovascular effects similar to those of intravenously infused dopamine. Plasma levels and urinary excretion of metabolites were investigated in dogs after oral administration of 4 mg/kg of ibopamine hydrochloride. Epinine, which was readily formed from ibopamine by esterases hydrolysis, was present in plasma in free and sulphate-conjugated form. The urinary metabolites after 6 h from the administration amounted to 62% of the dose, as a sum of 37% of epinine 3-O-sulphate, and 15 and 10% of 4-hydroxy-3-methoxyphenylacetic acid and 3,4-dihydroxyphenylacetic acid, respectively, both in free and conjugated form. When the main metabolite, epinine 3-O-sulphate, was administered intravenously it appeared to be excreted in urine without being deconjugated to any detectable extent, while it appeared to be partially deconjugated on oral administration.

Changes in vessel ultrastructure during ischemia and reperfusion of rabbit hindlimb: implications for therapeutic intervention.

Peripheral ischemia was induced in the rabbit by occlusion of the left iliac artery for 6 hr, followed by 24 hr of reperfusion. Biochemical and morphological investigations were performed to evaluate the extent of vascular and tissue injury. Blood samples for plasma enzyme determinations (creatine kinase (CK) and lactate dehydrogenase (LDH) activities) were obtained at times t = 0, t = 6, t = 30 hr. Plasma CK and LDH activities in ischemic animals were approximately twice as high as those in sham-operated animals at the end of reperfusion, although no difference was observed at the end of the period of ischemia. Morphological and morphometric analysis of extensor digitorum longus muscle from ischemic animals showed a reduction in the number of patent capillary vessels per muscle fiber (1.54 +/- 0.1 and 1.04 +/- 0.09, P < 0.05, in sham and ischemic groups, respectively; mean +/- SEM). In addition, the number of microvilli on endothelial surfaces were considerably increased in the ischemic group (0.14 +/- 0.02 and 0.41 +/- 0.01 microns -2, P < 0.001, in sham and ischemic groups, respectively). A great number of adhered leucocytes were found on the vessel surface with some leucocytes having migrated through the vessel wall. Microcirculatory damage was accompanied by the formation of microthrombi which sometimes occluded the entire vessel lumen. The infusion of 1 mg/kg/hr of cloricromene for 6 hr prevented ischemic injury in microvessels and also prevented swelling of muscle mitochondria. In the treated group the number of patent capillaries per muscle fiber was very similar to that found in sham-operated animals (1.49 +/- 0.08; P < 0.01 vs. ischemic control). In conclusion, several different cell types are involved in the pathophysiological changes which occur in microvessels during ischemia/reperfusion injury. Pharmacological interventions, which inhibit the interactions of blood cells with endothelium, may be of value in the treatment of peripheral ischemia/reperfusion injury.

Cystometric changes in alloxan diabetic rats: evidence for functional and structural correlates of diabetic autonomic neuropathy.

Autonomic neuropathy and urinary bladder function were compared in Sprague-Dawley rats with alloxan-diabetes of 3 months duration, rats fed sucrose for 8 weeks, and rats examined 8 weeks after pelvic nerve surgical axotomy; normal age-matched rats were used as controls. All experimental interventions induced bladder hypertrophy with increased bladder weight. In diabetic and sucrose-fed animals, water intake and urinary output increased. Cystometric recordings of normal rats in vivo showed rhythmic contractions (1.25 +/- 0.25 contr/min) with threshold volume for micturition reflex at 0.51 +/- 0.04 ml. In diabetic rats, bladder contractions were irregular and of lower frequency (0.60 +/- 0.04 contr/min), while threshold volume was significantly higher (1.00 +/- 0.11 ml). Bladder contractions were normal in sucrose-fed animals, though threshold volume was markedly augmented (1.27 +/- 0.19 ml). Pelvic nerve surgical ablation abolished micturition reflex. In bladder strips excised post-mortem, contractile response to field stimulation was reduced in diabetic rats compared to control and sucrose-fed animals. Morphological examination of pelvic and hypogastric nerves revealed abnormalities characteristic of diabetic neuropathy only in diabetic rats. These data suggest that in alloxan-induced diabetes the decrease in the rate of bladder contraction is the result of autonomic neuropathy; while bladder hypertrophy in sucrose-fed rats appears to be an organ adaptation to hyperdiuresis.

Urinary bladder dysfunction in the BB/W diabetic rat: effect of ganglioside treatment on functional and structural alterations.

Urinary bladder dysfunction in the diabetic BB/W rat is characterized by infrequent irregular contractions of high amplitude. Initially these occur in the absence of detectable neuroanatomical lesions of sensory afferent and parasympathetic fibers of the pelvic nerve, which constitute the micturition reflex arc. Structural lesions consisting of progressive axonal atrophy of myelinated and unmyelinated fibers become detectable only after 4 months of diabetes. In the current study we evaluated the effect of ganglioside treatment (10 mg./kg. body weight) for one month. This drug regimen was initiated at 4 months of diabetes, when functional bladder abnormalities were well established, whereas structural lesions were yet to appear. Animals examined 1 or 3 months after termination of the one-month treatment protocol showed sustained normalization of the characteristic functional abnormalities, accompanied by prevention of the neuroanatomical lesions of sensory afferent and parasympathetic efferent myelinated fibers in the pelvic nerve. These data suggest that ganglioside treatment may be beneficial in delaying the progression of diabetic autonomic neuropathy in this experimental animal model.