Intravesical repair of vesicovaginal fistula guided by cystoscopy.

BACKGROUND: Vesicovaginal fistulas (VVF) are an unusual problem that may significantly affect a patient's quality of life. The main causes for this condition are labour complications (mostly in developing countries) and pelvic surgeries (in industrialised countries). Treatment may be conservative or surgical. Regarding surgical treatment, there is still debate about the best approach and surgical technique. OBJECTIVE: To demonstrate a correction of a VVF guided by cystoscopy using intravesical laparoscopic instruments. METHODS: Case report and surgical video of a recurrent VVF treated with a hybrid technique involving direct transvesical insertion of 3 mm laparoscopic trocars and instruments guided by cystoscopy. As far as we know, although there are some reported techniques that use a combination of transvesical laparoscopic instruments and cystoscopy, this is the least invasive and most ergonomic technique described. RESULTS: Two years after surgery, the patient remains asymptomatic and with no fistula recurrence. CONCLUSION: The transvesical approach guided by cystoscopy seems to be an effective, safe and ergonomic minimally invasive procedure for VVF repair.

Acetyl-L-carnitine provides effective in vivo neuroprotection over 3,4-methylenedioximethamphetamine-induced mitochondrial neurotoxicity in the adolescent rat brain.

3,4-Methylenedioximethamphetamine (MDMA, ecstasy) is a worldwide abused stimulant drug, with persistent neurotoxic effects and high prevalence among adolescents. The massive release of 5-HT from pre-synaptic storage vesicles induced by MDMA followed by monoamine oxidase B (MAO-B) metabolism, significantly increases oxidative stress at the mitochondrial level. l-Carnitine and its ester, acetyl-l-carnitine (ALC), facilitate the transport of long chain free fatty acids across the mitochondrial membrane enhancing neuronal anti-oxidative defense. Here, we show the potential of ALC against the neurotoxic effects of MDMA exposure. Adolescent male Wistar rats were assigned to four groups: control saline solution, isovolumetric to the MDMA solution, administered i.p.; MDMA (4x10 mg/kg MDMA, i.p.); ALC/MDMA (100 mg/kg 30 min of ALC prior to MDMA, i.p.) and ALC (100 mg/kg, i.p.). Rats were killed 2 weeks after exposure and brains were analyzed for lipid peroxidation, carbonyl formation, mitochondrial DNA (mtDNA) deletion and altered expression of the DNA-encoded subunits of the mitochondrial complexes I (NADH dehydrogenase, NDII) and IV (cytochrome c oxidase, COXI) from the respiratory chain. Levels of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) were also assessed. The present work is the first to successfully demonstrate that pretreatment with ALC exerts effective neuroprotection against the MDMA-induced neurotoxicity at the mitochondrial level, reducing carbonyl formation, decreasing mtDNA deletion, improving the expression of the respiratory chain components and preventing the decrease of 5-HT levels in several regions of the rat brain. These results indicate potential benefits of ALC application in the prevention and treatment of neurodegenerative disorders.

Effects of prenatal cocaine exposure in the photoreceptor cells of the rat retina.

Despite the increasing evidence of eye abnormalities, the effects of prenatal exposure to cocaine on the visual system are still poorly understood. This study was aimed at analyzing the qualitative and quantitative organization of the retinal photoreceptor cells (PR) and outer nuclear layer (ONL) after prenatal exposure to cocaine in the rat. Pregnant Wistar rats were given sc injections of cocaine hydrochloride (60 mg/kg body wt/d) or saline or were not manipulated; analyses were performed in the 14- and 30-d-old male offspring. Radial semithin and ultrathin sections of epon-embedded flat mounts of the retina showed displaced PR-like cells in the inner nuclear layer (INL), picnotic PR nuclei in INL, and ONL, and retinal PR rosettes and outer-segment debris in the subretinal space. The quantitative study showed an increased density of PR-like nuclei in the INL in PND14 cocaine-treated rats that were within normal values at PND30; no changes were detected in the PR mean nuclear diameter and in the packing density of PR nuclei in the ONL. These data constitute the first morphological demonstration of photoreceptor damage after prenatal cocaine-exposure probably owing to a direct action of the drug and/or to the cocaine-induced ischemia/hypoxia.

Effects of prenatal cocaine exposure in the prefrontal cortex of the rat. A morphometric evaluation.

This work was undertaken in order to assess the organization of the prelimbic area of the medial prefrontal cortex of rats exposed prenatally to cocaine. Pregnant Wistar rats were assigned to the following groups: 1. Cocaine--60 mg/kg body wt/d sc, from gestational days 8-22; 2. Saline; 3. Pair-fed; and 4. Nonmanipulated. Male offspring were perfused on postnatal days 14 and 30. Six brains per group and per age were embedded in celloidin to calculate the volumes of the prelimbic area; sections from the other six brains were embedded in resin and processed for electron microscopy. Using semithin sections (2 microns) of layers II-III and V-VI, the following parameters were calculated: 1. The fraction of the neuropil occupied by neurons (VV); 2. The packing (NA) density; and 3. The numerical (NV) density. Qualitative alterations consisted of dispersed profiles of degenerated neurons and dendrites in the medial prefrontal cortex. No significant differences were found in the gross morphometric parameters when the cocaine group was compared with the other groups. A high interanimal variation was shown in the prelimbic volumes of postnatal day (PND) 14 cocaine-treated rats, and a a decrease in volumes was detected at PND30. Although there are some alterations in the main afferent cortical target area for dopaminergic input, its gross morphometric parameters do not seem to be sufficiently affected to account for the behavioral alterations referred to as being dependent on this brain region.

Effects of prenatal cocaine exposure in the retinal ganglion cell layer of the rat. A morphometric analysis.

To study the effects of prenatal cocaine-exposure on the developing retinal ganglion cell layer of the rat, female Wistar rats were administered subcutaneously (sc) cocaine hydrochloride (60 mg/kg body wt/d) or saline, or were not manipulated from gestational d 8-22. Male offspring were sacrificed at postnatal day 14 and 30. Radial semithin sections of epon-embedded flat mounts of the retinal quadrants were used to evaluate the following parameters along the centroperipheral axis: 1. Thickness of ganglion cells plus nerve fiber layer; 2. Nuclear size of ganglion cell layer neurons; and 3. Linear density (number per unit length) of ganglion cell layer neurons. To study the effects of cocaine and age on the retinal areas (temporal/nasal, dorsal/ventral), a repeated measures analysis of variance was used for each of the parameters mentioned above. All parameters were affected by prenatal exposure to cocaine. The thickness of the ganglion cell plus nerve fiber layer was reduced in cocaine-exposed rats in comparison with the saline group. Nuclear diameters were smaller in the cocaine than in the saline and control groups. The linear density was higher in the cocaine-exposed group than in the control and saline groups. The age-dependent decrease in the linear density from postnatal day 14-30 was higher in the cocaine-exposed rats than in the saline group; the decrease in the linear density along the centroperipheral axis found in both the control and saline groups was not significant in the cocaine-treated group. These morphometric findings strongly support the view that prenatal cocaine-exposure induces marked changes in the organization of the developing retina.

Methamphetamine exacerbates the toxic effect of kainic acid in the adult rat retina.

The recreational use of the psychoactive drug, methamphetamine has increased markedly over the last three decades. It has long been known that this drug has detrimental effects upon the mammalian brain monoaminergic system, but the long- or short-term effects on the retina, a neurological extension of the central nervous system, have received little attention. The aim of this study was, therefore, to determine whether intraocular injection of methamphetamine (MA) is toxic to the healthy adult rat retina and to analyse its effects on the compromised retina after an injection of the ionotropic glutamate receptor agonist, kainate, which is known to cause retinal neuropathology. The equivalent of 1 mM (in the vitreous humour) MA and/or kainate (40 microM) were injected intravitreally. Flash electroretinograms (ERGs) were recorded before and 2 and 4 days after treatment. Five days after treatment, animals were killed and the retinas analysed either for the immunohistochemical localisation of various antigens or for electrophoresis/Western blotting. Some animals were kept for 19 days after treatment and the retinas analysed for tyrosine hydroxylase immunoreactivity. No differences could be found between vehicle- and MA-treated retinas with respect to the nature or localisation of either tyrosine hydroxylase immunoreactivity after 5 or 19 days or other antigens after 5 days. Moreover, the normal ERG and GFAP and calretinin protein antigens were unaffected by MA. Kainate treatment, however, caused a change in the ERGs after 2 and 4 days, an alteration in every antigen localised by immunohistochemistry and an increase in the retinal levels of calretinin and GFAP proteins. Significantly, the changes seen in the b-wave amplitude and implicit time of the ERG after 4 days and the increased level of GFAP protein after 5 days following kainate treatment were enhanced when MA was co-injected. Intravitreal injection of methamphetamine had no detectable detrimental effect on the normal adult rat retina but exacerbated the damaging effects of kainic acid. Such data suggest that a neurotoxic effect of MA may be more obviously illustrated when the tissue is already compromised as occurs in, for example, ischemia.

Differential effects of prenatal exposure to cocaine and amphetamine on growth parameters and morphometry of the prefrontal cortex in the rat.

The purpose of this study was to investigate the differential effects of prenatal exposure to psychostimulants, e.g., cocaine or amphetamine, on basic growth parameters and morphometry of the medial prefrontal cortex of the rat. A group of pregnant Wistar rats was given 60 mg/kg body weight/day of cocaine hydrochloride and another group 10 mg/kg body weight/day of d-amphetamine sulfate, subcutaneously, from gestational days 8 to 22. Control groups of pregnant rats were pair-fed; litters were culled to eight pups (4 males and 4 females) weighed every other day until postnatal day 30 and every week until day 90. The body weight growth patterns modelled by a Gompertz curve were different in rats prenatally exposed to the two psychostimulants. Rats exposed to amphetamine had on average a slower growth than those exposed to cocaine, reaching an identical estimated adult weight. Allometric relationships between forebrain and body weight and cerebellum and body weight were described by two distinct postnatal growth phases that are different among the experimental groups. In the comparison of the two psychostimulants the relative cerebellum/body growth is lower in the offspring of the cocaine group than in the amphetamine group between PND14-PND30; between PND30-PND90 the relative growth rate is considerably higher in the offspring of the cocaine dams compared to that of the amphetamine dams. Groups of perfused animals were selected at postnatal days 14 and 30 to analyze the morphometric organization of the medial prefrontal cortex. In serial celloidin sections the volumes of the prefrontal cortex were determined; the number of neurons per unit volume of reference area was calculated using the stereological technique of the disector. The changes found in the morphometric parameters show a catch-up at postnatal day 30 of the "increased" density of neurons of the medial prefrontal cortex found at postnatal day 14. These data show differential growth patterns of offspring from cocaine- and amphetamine-exposed rats; a delayed development in the achievement of normal morphometric parameters of neurons in the prelimbic subarea of the medial prefrontal cortex occurs in the prenatally amphetamine-exposed offspring at early ages, and a catch-up is found after the first month of life. Complementary studies are needed to assess whether these changes have functional implications in the rats exposed prenatally to psychostimulants.

Development of the eye after gestational exposure to cocaine. Vascular disruption in the retina of rats and humans.

Clinical and basic research in the area of drugs of abuse are of utmost importance since they provide the necessary background for health programs in one of the main problems of contemporary society. The available data in this field demonstrate that acute, subacute and/or chronic abuse of illicit drugs, e.g., cocaine, alters the neurochemistry and functioning of the neural circuitries. Although recent works demonstrated that the visual system is lesioned after exposure to cocaine during the active periods of development, no studies have provided detailed information on the effects of these substances on the development of this sensory system. The present paper will report: 1) the vulnerability of the developing visual system to gestational exposure to cocaine; 2) the effects of cocaine in the visual system during the more active periods of development in humans and, as far as possible, the establishment of homologies with animal models where exposure is made in corresponding periods of human gestation, and 3) the characterization of the vascular disruption caused by ischemic/hypoxic mechanisms. The clinical study focused the ophthalmologic evaluation of newborns exposed in utero to illicit drugs. Newborns exposed to cocaine in utero showed marked vascular disruption in the retina: superficial and deep hemorrhages that, although morphologically similar to neonatal retinal hemorrhages, presented a longer reabsorption time when compared with the neonatal hemorrhagic lesions due to birth trauma in the general population. Prolonged eyelid edema was also a prominent finding. The animal study was conducted in Wistar rats exposed prenatally (gestational days 8 to 22) and postnatally (postnatal days 1-6, 1-13 and 1-29) to 60 mg/kg body weight/day and 15 mg/kg body weight/day, respectively, to cocaine hydrochloride administered subcutaneously; control groups included pair-feeding during the same experimental periods. Similar alterations to those observed in the newborns where exposure to cocaine was affirmative, were found: intraretinal hemorrhages allied to signs of chronic ischemia both in the outer retina-photoreceptor rosettes and in the inner retina-epiretinal glial membranes. Taking into consideration that the visual system is one of the more important sensory systems, the identification and characterization of these alterations, the similarity between animal and human findings, and their relation with cocaine per se, can provide a sound data base for illicit drug prevention programs.

Developmental exposure to methamphetamine: a neonatal model in the rat.

Taking into account that methamphetamine (MA) is a popular recreational drug among young adult women, i.e., in gestational age, the present model aims to assess the effects of its exposure during development. In this experimental model, MA effects are assessed in the rat during the first month of life, regarding both general growth parameters, and gross morphological effects in the retina as part of the evaluation of sensory systems. Experimental animals were obtained from 60-day-old nulliparous females. Litters were culled to 8 pups (4 males and 4 females, whenever possible), individually marked and weighed every two days. Experimental groups received 10 mg (+)methamphetamine hydrochloride kg body weight/day, subcutaneously, twice daily, from postnatal day (PND) 1 to the day before sacrifice; control groups received isovolumetric doses of saline, in the same protocol. Pups were weaned on PND 21. Groups were sacrificed on PND 5, 7 and 30. Animals exposed to MA presented increased percentage of retinal hemorrhages (18, 7 and 11% on PND 5, 7 and 30, respectively) compound to control groups (2% on PND 7, 0% on PND 5 and 30). On PND 30, the mean body weight of males exposed to MA was 75% of the mean weight of male controls, whereas for females, mean body weights were 70% of those of female controls. These findings support the view that developmental parameters in the rat are deleteriously affected by early exposure to MA.

Effects of neonatal exposure to cocaine in the development of the neurotransmitters retinal systems: an immunocytochemical and neurochemical study.

The visual system of rodents is affected if exposure to drugs, e.g., cocaine, occurs during prenatal or early postnatal development. This study aims to evaluate, in an experimental model of neonatal exposure to cocaine in the rat, the immunocytochemical expression of tyrosine hydroxylase in the retina and the levels of different neurotransmitters and its metabolites. Male Wistar rats were given 15 mg cocaine hydrochloride/kg body weight/day, subcutaneously, in two daily doses, from the day after birth (PND1) to PND6, 13, and 29. Controls were given 0.9% saline. Groups of rats were perfused at PND7, 14, and 30 with fixative, and the retinas were processed as wholemounts, and immunostained with the antibody anti-TH. Other groups were decapitated, and the retinas were dissected and processed by high performance liquid chromatography with electrochemical detection (HPLC-EC) for determination of dopamine and metabolites (DOPAC and HVA). A reduction in the retinal surface area was detected in the PND30 cocaine group, and a decrease in the density of the small TH-IR cells was found in the PND14 cocaine group although not reaching significant levels. The other quantitative parameters did not differ between the control and cocaine groups. The levels of neurotransmitters did not significantly differ between the groups at any age. These results show a differential vulnerability of the dopaminergic system of rats exposed neonatally to cocaine when compared with the effects found after prenatal exposure to the same drug.

Neonatal methamphetamine in the rat: evidence for gender-specific differences upon tyrosine hydroxylase enzyme in the dopaminergic nigrostriatal system.

Methamphetamine (Meth) neurotoxicity upon the mesencephalic dopaminergic systems was demonstrated in the adult, both in humans and in experimental models. In the rat, the development and maturation of the dopaminergic systems is accomplished during the first month of postnatal life, a period of particular vulnerability to environmental influences. In this study, the effect of Meth exposure during the first month of life was assessed in the nigrostriatal dopaminergic system of the rat. For this purpose, Wistar rat litters were culled to 8 pups, retaining preferentially 4 males and 4 females, which, in the day following birth (postnatal day 1, PND1), were randomly attributed to either the Meth or control group. Meth-groups were administered 10 mg of (+)-methamphetamine hydrochloride/kg body weight/day, subcutaneously, twice daily, from PND1 until PND29; control groups received isovolumetric doses of saline. Animals were sacrificed at PND30. Males exposed to Meth during the first month of life had increased tyrosine hydroxylase (TH) activity both in the caudate-putamen and substantia nigra. Males also had increased nigral TH mRNA levels, as assessed by in situ hybridization. These effects did not exist in females. These results support the evidence that Meth exposure during the first month of life in the rat has a gender-specific stimulatory effect upon the maturation of TH, the key enzyme for dopamine biosynthesis in the nigrostriatal dopaminergic system.

Body weight gain and hippocampal volumes of rats exposed neonatally to psychostimulants.

There is increasing evidence that early exposure to psychostimulants exerts long-lasting effects on the central nervous system. Yet, analysis of the body weight gain and volumetric determinations of brain areas have not been performed by comparing the effects of neonatal exposure to cocaine and amphetamine. Male (Wistar) rats were given cocaine hydrochloride (15 mg/kg body weight/day), D-amphetamine sulphate (25 mg/kg body weight/day) or saline, s.c., twice daily, from postnatal day (PND) 1 to 30. The experimental design used random permuted blocks of 4 males per litter -9 litters for body weight gain evolution and 9 for the analysis of body, brain and cerebellum at PND30. Volumes of the hippocampal formation were calculated in horizontal serial sections of celloidin embedded material from 6 animals per group. The analysis of body weight gain evolution was performed using discriminant functions and allowed the separation of the amphetamine group from the remainder and the control from the psychostimulants group; weight gain in PND 24-30 period presented the highest discriminating power. The mean volume of the hippocampal formation was lower in the psychostimulants group, and the differences were significant in the molecular layer of the dentate gyrus of cocaine and amphetamine exposed animals when compared with the controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Remodeling of the cerebellar glomeruli after long-term alcohol consumption in the adult rat.

The morphological effects of the prolonged alcohol consumption on the cerebellar glomeruli of adult rats were studied in groups of controls and animals fed alcohol for 1, 3, 6, 12 and 18 months, by applying qualitative and quantitative ultrastructural methods. Following 6 months of alcohol consumption degenerated granule cell dendritic profiles were randomly dispersed over the granular layer surrounding mossy fiber terminals. In 12- and 18-month alcohol-fed groups, some glomeruli appeared with an atrophic design owing to the lack of their post-synaptic targets while others presented a remodeled organization, with Golgi cell dendrites replacing the missing granule cell digits. An extensive glial reaction was seen investing both types of glomeruli. The total number of synapses per glomerulus remained unchanged in all groups. This is due to an increase of the mossy fiber terminal--Golgi cell dendrite synapses, which compensate the decrease of the mossy fiber terminal--granule cell dendrite synapses. It is suggested that the morphological remodeling of the cerebellar glomeruli after long-term alcohol consumption can lead to changes in the balance of the excitatory-inhibitory activities of the cerebellar circuitry. This could be related to the important functional changes observed in the cerebellum under these circumstances.

Long term alcohol consumption induces microtubular changes in the adult rat cerebellar cortex.

The effects of prolonged alcohol consumption on the microtubules of Purkinje cell dendrites and granule cell axons were studied in adult rats fed alcohol for 1, 3, 6, 12 and 18 months and compared with respective age-matched controls. A significant consequential decrease in the number of dendritic microtubules in alcohol-fed rats was found when compared with the respective controls. Conversely, an increase in the number of these organelles was found in both ascending and parallel portion of the axons in the experimental animals. The possibility of a relationship between microtubular changes and previously reported cerebellar cortex alcohol-induced structural alterations is advanced.

A quantitative study of frontal cortex dendritic microtubules in patients with Alzheimer's disease.

The dendritic microtubules (MT) of the frontal cortex layers II and III were studied in 9 patients with Alzheimer's disease (AD) and the results compared with 9 case controls. Dendrites with abnormally oriented MTs and others depleted of these structures were seen. A significant reduction in the number of MTs per unit area was found in AD. It is suggested that microtubular changes in AD can interfere with neuroplasmic transport and thus, be implicated with the dendritic degeneration present in this disease.

Granule cell loss and dendritic regrowth in the hippocampal dentate gyrus of the rat after chronic alcohol consumption.

The effects of chronic alcohol consumption (CAC) on the relative number of dentate gyrus granule cells and their dendritic trees, were studied in animals fed alcohol for 6, 12 and 18 months and in their respective controls. The granule cell density was estimated with the unbiased disector method. Following 6 months of alcohol consumption, the thickness of the dentate gyrus granular layer and the relative number of dentate granule cells were significantly decreased when compared with controls. The granule cell dendritic arborizations showed an increase of their dendritic extent in alcohol-treated rats. No significant differences were found in the density of dendritic spines between alcohol-fed and control animals. These results indicate the existence of hippocampal granule cell dendritic regrowth in alcohol-fed rats, probably occurring as a compensatory response to the granule cell deficit which follows the alcohol-induced granule cell degeneration. These degenerative and regenerative changes might have functional implications for the organization of the synaptic hippocampal circuitry after long periods of alcohol consumption.

Effects of prenatal exposure to amphetamine in the medial prefrontal cortex of the rat.

This study was designed to investigate the effects of prenatal exposure to amphetamine in the organization of the medial prefrontal cortex of the rat, by an evaluation of growth, morphometric and neurochemical parameters. Pregnant Wistar rats were given 10 mg/kg body weight/day of D-amphetamine sulfate, subcutaneously, from gestational days 8 to 22. Control groups of pregnant rats were injected with saline, pair-fed or non-manipulated; litters were culled to eight pups (four males and four females), weighed every other day until postnatal day 30 and every week until day 90. The Gompertz model was used to study body weight evolution and the estimated growth parameters were not significantly different in the experimental groups. At postnatal days 14 and 30, the volumes of the prefrontal cortex, the fraction of neuropile occupied by neurons and the number of neurons per unit surface are were determined. The number of neurons per unit volume of reference area was calculated using the stereological technique of the dissector. For neurochemical analysis, the medial prefrontal cortex was dissected to measure the concentration of dopamine, serotonin and their metabolites. The allometric relationship of forebrain/body growth pointed to a mechanism of sparing and compensatory growth in the amphetamine exposed group. The changes found in the number of neurons per unit volume at postnatal day 14 show a catch-up at postnatal day 30. A decrease in serotonin levels was found in the amphetamine group compared with the pair-fed control, which was reflected in the ratio of serotonin to its metabolite, 5-hydroxyindolacetic acid. These changes, whether permanent or transitory, raise the possibility that some of the effects of prenatal exposure to amphetamine may be due to modifications in the neurotransmitter levels of serotonin.

Alcohol withdrawal does not impede hippocampal granule cell progressive loss in chronic alcohol-fed rats.

Using the unbiased disector method we have shown that chronic alcohol consumption induces a significant decrease of dentate gyrus granule cell density in alcohol-fed rats for 18 months. A still more dramatic reduction was observed in a group of age-matched rats alcohol-fed for 12 months and switched to water for 6 months (recovery group). These results indicate that a progressive neuronal loss of the hippocampal granule cells in not impeded after cessation of alcohol intake. It is thus suggested that once the mechanisms underlying the alcohol-induced neuronal degeneration are triggered, they continue to act even after withdrawal.

Dendritic spine plasticity and chronic alcoholism in rats.

Alcohol consumption in rats leads to degenerative changes in the cerebellar cortex (described elsewhere). Purkinje cells show a loss of dendritic spines, but some of the intact spines elongate to 3-4 times their normal length. This is postulated as a plastic change resulting from the degeneration of the original axon terminal contact with a consequent elongation of the spine 'in search of' and finally synapsing with still viable parallel fibre terminals in the vicinity. Under these conditions the circuitry is radically altered.

Giant multivesicular bodies in the rat hippocampal pyramidal cells after chronic alcohol consumption.

Multivesicular bodies (MVBs) with diameters up to 4.5 microns were observed in the hippocampal pyramidal cells of rats submitted to chronic alcohol consumption. A significant increase in the volumetric density (Vv) of these organelles was found in CA1 pyramidal cells. Transitional forms of MVBs towards lysosomes were seen. A failure in MVB's enzymatic hydrolytic mechanisms, due to the prolonged alcohol aggression, could underlie its formation.