Quantitative immunohistochemical analysis of nitric oxide synthases and apoptosis regulator proteins in the fetal rat brain following maternal uterine artery ligation.

The purpose of this study was to determine the relation between nitric oxide synthases (calcium-independent iNOS and calcium-dependent eNOS) and apoptosis regulator proteins (anti-apoptotic Bcl-2, pro-apoptotic p53) of fetal rat brain in experimental intrauterine growth retardation (IUGR) model via quantitative immunohistochemistry. Cortical zone of parietal cerebral cortex and ventricular zone of third ventricle were studied following bilateral uterine artery ligation on gestational day 18. Significant increase in iNOS immunoreactivity was determined in parietal cerebral cortex and ventricular zones as eNOS immunoreactivity increased in ventricular zone of IUGR group. Bcl-2 expression was significantly decreased in ventricular zone; whereas cortical zone of IUGR group expressed p53 immunoreactivity.

Constructive effect of exogenous melatonin against osteoporosis after ovariectomy in rats.

OBJECTIVE: To analyze histomorphometric, densitometric and biochemical effects of melatonin on osteoporosis in ovariectomized rats. STUDY DESIGN: Wistar rats were divided into 6 groups. Group C: control; Group I: bilateral ovariectomy (OVX); Group II: OVX + vehicle; Group III: OVX + 10 mg/kg/day melatonin (MLT); Group IV: OVX + 30 mg/kg/day MLT; Group V: sham + 10 mg/kg/day MLT. Cortex, trabecula, osteoblast and osteoclast numbers were evaluated on vertebra and femur histomorphometrically. Hydroxyproline analysis was used to determine collagen content of femur and vertebrae. Bone mineral density and bone mineral content were measured. RESULTS: Trabecular thickness and trabecular area of vertebra and femur and cortical thickness of femur showed remarkable decrease after OVX, but increased after MLT treatment in the OVX+MLT groups. Following OVX, no statistically significant difference was found in number of osteoblasts or osteoclasts, trabecular number or levels of hydroxyproline after treatment with MLT. OVX caused significant decrease in bone mineral density, but treatment with MLT was unable to reverse this effect. CONCLUSION: MLT may trigger microscopic changes in bone, and time of application is critical for clinical recovery. It can be effective in helping treat postmenopausal osteoporosis. However, it is contraindicated in women who have normal-functioning ovaries.

Neuroprotective effects of melatonin upon the offspring cerebellar cortex in the rat model of BCNU-induced cortical dysplasia.

Cortical dysplasia is a malformation characterized by defects in proliferation, migration and maturation. This study was designed to evaluate the alterations in offspring rat cerebellum induced by maternal exposure to carmustine-[1,3-bis (2-chloroethyl)-1-nitrosoure] (BCNU) and to investigate the effects of exogenous melatonin upon cerebellar BCNU-induced cortical dysplasia, using histological and biochemical analyses. Pregnant Wistar rats were assigned to five groups: intact-control, saline-control, melatonin-treated, BCNU-exposed and BCNU-exposed plus melatonin. Rats were exposed to BCNU on embryonic day 15 and melatonin was given until delivery. Immuno/histochemistry and electron microscopy were carried out on the offspring cerebellum, and levels of malondialdehyde and superoxide dismutase were determined. Histopathologically, typical findings were observed in the cerebella from the control groups, but the findings consistent with early embryonic development were noted in BCNU-exposed cortical dysplasia group. There was a marked increase in the number of TUNEL positive cells and nestin positive cells in BCNU-exposed group, but a decreased immunoreactivity to glial fibrillary acidic protein, synaptophysin and transforming growth factor beta1 was observed, indicating a delayed maturation, and melatonin significantly reversed these changes. Malondialdehyde level in BCNU-exposed group was higher than those in control groups and melatonin decreased malondialdehyde levels in BCNU group (P<0.01), while there were no significant differences in the superoxide dismutase levels between these groups. These data suggest that exposure of animals to BCNU during pregnancy leads to delayed maturation of offspring cerebellum and melatonin protects the cerebellum against the effects of BCNU.

Light, electron microscopic and immunohistochemical study of the effect of low-dose aspirin during the proestrus phase on rat endometrium in the preimplantation period.

OBJECTIVE: To evaluate structural alterations in rat endometrium at preimplantation following treatment with aspirin beginning from proestrus by light microscopy, electron microscopy and immunohistochemical techniques. STUDY DESIGN: Twenty rats were divided into control (n = 10) and experimental (n = 10) groups. Experimental rats were treated with low-dose aspirin daily (2 mg/kg/day) during estrus, beginning from the proestrus phase, mated at end of cycle and treated with aspirin. Untreated pregnant rats were the control group. Rats in both groups were sacrificed at the 84th pregnancy hour; the uterus was rapidly removed and dissected free of surrounding adipose tissue. Uteri specimens from nonpregnant rats were transferred into fixative solution and processed for light, electron microscopic and immunohistochemical study. RESULTS: Light and electron microscopy of endometrium from control rats conformed to mid-diestrus phase; endometrial histology of the aspirin-treated group conformed to late diestrus phase. The endometrial layer was significantly thicker in the aspirin-treated group compared to the untreated control group (p <0.001). No significant difference was found in vessel number between groups. Staining with alphaV integrin was more dense in the aspirin-treated group. CONCLUSION: Based on histologic findings, we suggest low-dose aspirin has positive effects on preparing endometrium before implantation.

Efficacy of melatonin on offspring liver maturation in pinealectomized pregnant rats subjected to experimental epilepsy.

BACKGROUND AND AIMS: In clinical practice, maternal epilepsy is a disabling disease for newborn infants, but current data concerning the effect of epileptic phenomena in pregnant mothers on newborns are still limited. This study was undertaken to investigate the effects of pinealectomy (Px) and melatonin treatments on the morphological changes in the liver tissue of newborn rats following experimental epilepsy during pregnancy. METHODS: Female Swiss Albino rats were divided into five groups: intact control group; saline control group; epilepsy group; epilepsy plus Px group; and melatonin-treated epilepsy plus Px group. At one month after Px, an acute grand mal epileptic seizure was induced by penicillin-G during their pregnancy in all animals except the control groups. On the neonatal first day, newborn rats were perfused with intracardiac fixative solution, and then livers were removed and processed for toluidine blue, periodic acid-Schiff (PAS) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) reactivity. RESULTS: Normal migration and hepatic maturation were determined in the postnatal rat liver in the control groups, while the morphological structure of the liver in the epilepsy and epilepsy plus Px groups corresponded to the early embryonic period. In the melatonin-treated epilepsy plus Px group, the number of TUNEL positive cells decreased significantly compared to both epilepsy and epilepsy plus Px groups; however, there was no statistically significant difference from the control groups as a result of melatonin activity. CONCLUSIONS: Some histological findings consistent with chronic fetal distress in newborns of mother rats with epilepsy and Px were observed. Melatonin could be a candidate protective drug for the development of liver tissue in pregnant patients with epilepsy.

Immunohistochemical profile of transforming growth factor-beta1 and basic fibroblast growth factor in sciatic nerve anastomosis following pinealectomy and exogenous melatonin administration in rats.

Collagen scar formation at the cut end of a peripheral nerve, an important problem in clinical practice for neurosurgeons, obstructs sprouting of axons into appropriate distal fascicles, and thereby limits the regeneration process. Researchers have attempted to control collagen accumulation and neuroma formation with various physical and chemical methods, but with limited functional success. Recently, it has been demonstrated that transforming growth factor (TGF)-beta and basic fibroblast growth factor (bFGF) play an important role in collagen production by fibroblasts and in Schwann cell activity. In our study, rats were divided into a control group, a melatonin-treated group, a surgical pinealectomy group, and a group treated with melatonin following pinealectomy. They then underwent a surgical sciatic nerve transection and primary suture anastomosis. At 2 months after anastomosis, the animals were sacrificed and unilateral sciatic nerve specimens, including the anastomotic region, were removed and processed for immunohistochemical study from two animals in each group. For each antibody, immunoreactivity was assessed using a semiquantitative scoring system. Strong TGF-beta1 and/or bFGF expression was observed in the epineurium of animals that underwent pinealectomy, but no or weak staining was observed in animals in the control and melatonin treatment groups. Based on these data, we suggest that both TGF-beta1 and bFGF have important roles in control of collagen accumulation and neuroma formation at the anastomotic site, and that the pineal neurohormone melatonin has a beneficial effect on nerve regeneration.

The effect of exogenous melatonin administration on trabecular width, ligament thickness and TGF-beta(1) expression in degenerated intervertebral disk tissue in the rat.

Intervertebral disk (IVD) degeneration, a complex pathological condition of varying origins, causes low back pain. Degenerative changes in IVD tissue affect the adjacent vertebral structure, resulting in a decreased vertebral trabecular width. It has been suggested that transforming growth factor-beta 1 (TGF-beta(1)) may have a role in the repair of connective tissue, as it occurs in the IVD degeneration process. In this study, we investigated the effects of exogenous melatonin (MEL) administration on vertebral trabecular width, ligament thickness and TGF-beta(1) expression in degenerated IVD tissue. Fifteen adult male Swiss Albino rats were divided randomly into three groups; nonoperated control, operated degeneration, and MEL treatment groups. In the operated degeneration and MEL treatment groups, cuts were made parallel to the end plates in the posterior annulus fibrosus at the fifth and tenth vertebral segments of the tail to induce IVD degeneration. In each group, TGF-beta(1) immunoreactivity and morphometry of vertebral trabecular width and anterior and posterior ligament thickness were evaluated. Histologically, disorganisation and irregularity of collagen fibres was seen in the degenerated (operated) IVD. Increased TGF-beta(1) expression in multinuclear chondrocytes was also observed as was decreased vertebral trabecular width. Importantly, the reduction of trabecular width observed in the operated degenerated group was reversed after MEL administration (p<0.0001). Similarly, TGF-beta(1) expression in multinuclear chondrocytes was dramatically increased after exogenous MEL application. Thus, there was a regression in histopathological changes after MEL treatment, with disk appearances similar to those of the control group. Based on our findings, we suggest that MEL activates the recovery process in the degenerated IVD tissue, possibly by stimulating TGF-beta(1) activity. This is the first report investigating the involvement of the pineal hormone MEL in the repair of rat IVD.

Pinealectomy stimulates and exogenous melatonin inhibits harmful effects of epileptiform activity during pregnancy in the hippocampus of newborn rats: an immunohistochemical study.

OBJECTIVES: Epilepsy during the pregnancy is an important problem in clinical practice for newborn individuals. Recently, it has been demonstrated that mothers' epileptic seizures have some harmful effects on newborns, but present data concerning the effects of epileptic phenomena in pregnant mothers on newborn pups are still limited. The current study was undertaken to investigate the morphological changes in the hippocampus of newborn pups of pinealectomized rats subjected to experimental epilepsy during pregnancy. METHODS: In this study, rats were randomly divided into four groups (ten animals each): intact control group, epilepsy control group, surgical pinealectomy + epilepsy group, and group with melatonin treatment following pinealectomy procedure. The animals in surgical pinealectomy + epilepsy and melatonin treatment groups underwent a surgical intervention consisting of pineal gland removal. At 1 month after surgical pinealectomy, an acute grand mal epileptic seizure was induced by 400 IU penicillin G administration into their hippocampal CA3 region on the 13th day of their pregnancy in all animals except the intact control animals. On the first neonatal day, the hippocampi were removed and processed for microscopic examination. Nestin expression was analysed in the developing hippocampal tissue. RESULTS: Normal migration and hippocampal maturation were determined in the postnatal rat hippocampus in intact control group, but the morphological structure of the hippocampus in the epilepsy control group corresponded to the early embryonal period. It was found that experimental epilepsy and pinealectomy enhanced nestin immunoreactivity, whereas exogenous melatonin treatment (30 mug/100 g body weight, intraperitoneal) inhibited pinealectomy-stimulated nestin expression in CA1 region of the hippocampus. CONCLUSION: These findings suggest that epileptic seizures during pregnancy may cause an impaired hippocampal neurogenesis and neuronal maturation in the newborn, and the negative effects in the postnatal rat hippocampus are more dramatic after pinealectomy of the mother; conversely, melatonin administration suppresses these negative changes. This is the first report investigating the effects of maternal epilepsy during pregnancy in pinealectomized rats on nestin immunoexpression in the newborn rat hippocampus.

Neonatal pinealectomy induces Purkinje cell loss in the cerebellum of the chick: a stereological study.

Melatonin plays an important role in certain physiological functions and morphological features of various structures. In the current study, the effects of pinealectomy on Purkinje cell number and morphological features of developing cerebellum in the chick were investigated using stereological methods. Fifteen Hybro Broiler newly hatched chicks were divided into three groups: a pinealectomized group (n = 5), sham-operated group (n = 5) and a non-pinealectomized control group (n = 5). Surgical pinealectomy was performed in 3-day-old chicks. In the 8th week, all animals were sacrificed for histopathological evaluation and subsequent stereological analysis. Each layer volume of molecular (+Purkinje cell), granular and white matter in the cerebellum was estimated in all animals. It was found that there was no significant difference for the volume of whole cerebellum and also molecular (+Purkinje cell) layer in these groups (P > 0.05). Nevertheless, the values of granular layer and white matter of sham-operated group were significantly different from those of control and pinealectomized animals (P < 0.01). It was also observed that pinealectomy significantly reduces the Purkinje cell number in cerebellar cortex (P < 0.01). The present study is the first stereological study to demonstrate the histomorphological effects of pinealectomy on the cerebellum in the chick. Our results suggest that pineal gland/melatonin might play an important role in morphological features of the developing cerebellum in the chick.

Beneficial effects of melatonin on morphological changes in postnatal cerebellar tissue owing to epileptiform activity during pregnancy in rats: light and immunohistochemical study.

Although it has been demonstrated that maternal epilepsy has some harmful effects on newborn individuals, current data concerning the effects of epileptic phenomena in pregnant mothers on newborn pups are still limited. This study was undertaken to investigate the changes in the cerebellum of newborns of pinealectomized rats subjected to experimental epilepsy during pregnancy. In our study, the rats were randomly divided into six groups: intact control group, anesthesia control group, epilepsy group, melatonin-treated epileptic group, surgical pinealectomy group, and group of melatonin treatment following pinealectomy procedure. At 1 month after pinealectomy, an acute grand mal epileptic seizure was induced by 400 IU penicillin-G administration into their intrahippocampal CA3 region during the 13th day of their pregnancy in all animals except intact control group. On the neonatal first day, pups were perfused transcardially and the cerebellums removed were processed for light microscopic and immunohistochemical studies. Normal migration and maturation were determined in the postnatal rat cerebellum in both intact control and anesthesia (ketamine-xylazine) control groups, but the morphological structure of cerebellum in the epilepsy control group corresponded to the early embryonal period. It was found that experimental epilepsy or pinealectomy procedure enhanced nestin immunoreactivity, but exogenous melatonin treatment (30 microg/100 g body weight, i.p.) following pinealectomy inhibited increased nestin expression induced by melatonin deprival in vermis region of newborn rat cerebellum (P < 0.001). Our results confirm that epileptic seizures during pregnancy may impair neurogenesis and neuronal maturation in newborns, which are more dramatic in the presence of melatonin deficiency during pregnancy, explaining more harmful effects of epileptic seizures to embryos of aged mothers. To the best of our knowledge, this is the first study reporting the effects of maternal epilepsy during pregnancy in pinealectomized rats on nestin immunoexpression in the newborn rat cerebellum.

Chronic changes in cerebrospinal fluid pathways produced by subarachnoid kaolin injection and experimental spinal cord trauma in the rabbit: their relationship with the development of spinal deformity. An electron microscopic study and magnetic resonance imaging evaluation.

Post-traumatic cystic changes in cerebrospinal fluid (CSF) pathways such as ventriculomegaly and/or hydrosyringomyelia are not uncommon, but their characteristics have not yet been fully clarified. This study was designed to investigate the alterations affecting the CSF pathways in rabbits at a late stage, and to clarify the relationship between these changes and the development of spinal deformity. In this study, a total of 60 New Zealand white rabbits were used and they were segregated into four different groups of 15 animals each: sham-operation group, kaolin group, and kaolin plus mild trauma group and kaolin plus severe trauma group. The animals were subjected to radiological investigation using direct X-ray study and magnetic resonance imaging (MRI) after 4 months. The thoracic spinal cords of the animals were dissected after intracardiac perfusion-fixation with 10% formalin for light microscopy and 2.5% glutaraldehyde for transmission electron microscopic study. Following the sectioning and staining procedures, the histological characteristics of the spinal cords were evaluated with light microscopy and transmission electron microscopy. A spinal deformity developed in 90% in rabbits in both kaolin injection group and spinal trauma groups. MRI revealed generalized dilatation of the ventricular system and the central canal of the spinal cord after the kaolin injection with/without trauma in this study. Gross morphologic examination showed some enlargement of entire CSF pathways in these groups. All animals with central canal dilatation had mild or severe scoliotic and kyphotic deformities. In a light microscopic study, a denuded ependymal line and multicyst formations in periependymal areas were found in both kaolin injection and spinal trauma groups. Ultrastructurally, an apical flattening of the ependyma, microcysts in the ependymal cells, axonal degeneration, demyelination, and loss of ependymal cells adjacent mild spongy were found in the spinal cords of animals in these groups. To the best of our knowledge, this is the first study to investigate the chronic effects of spinal cord injury (SCI) on the CSF pathways and their relationship with the development of spinal deformity in an experimental model of kaolin injection and trauma, using MRI as well as light and transmission electron microscopy. In the light of this study, the severity of spinal cord injury on the development of some degenerative findings in the spinal cord was clear, but further clinical and experimental studies using dynamic imaging techniques will be valuable.

Pinealectomy exaggerates and melatonin treatment suppresses neuroma formation of transected sciatic nerve in rats: gross morphological, histological and stereological analysis.

At present, an intensive effort for prevention of neuroma formation following peripheral nerve section continues. It has been recently suggested that surgical pinealectomy (Px) induces elevation of the collagen content in the granulation tissue of a wound, while melatonin application after Px suppresses elevation of the collagen accumulation in the tissue. The aim of the present study was to assess whether melatonin had the ability to suppress collagen production and neuroma formation following peripheral nerve transection. A total of 40 male rats (four groups of 10) were left intact (intact controls) or sham operated (sham group), were Px, or were Px and given melatonin (Px + melatonin group). All animals underwent a surgical intervention consisting of right sciatic nerve neurectomy. After 4 wk, the animals were killed following intracardiac perfusion. Gross morphology of neuroma formation in the proximal nerve segment was examined and proximal neuroma evaluated. Macroscopic and microscopic findings revealed that Px caused a proliferation of connective tissue and large neuroma formation at the proximal ends of transected nerves. Stereological analysis showed that there was a statistically significant reduction in connective tissue content of the same region in Px animals treated with melatonin (P < 0.005). The results achieved in a rodent model of sciatic nerve neuroma formation showed that there was a positive correlation between macroscopic and microscopic observations, and that melatonin enhanced axonal regeneration presumably due to its inhibitory effect on neuroma formation.

Influence of penicillin-induced epileptic activity during pregnancy on postnatal hippocampal nestin expression in rats: light and electron microscopic observations.

OBJECTS: Current data concerning the effects of maternal epileptic phenomena on newborns are limited. In clinical practice, therefore, it is difficult to suggest proper guidelines on this issue. This study was carried out to investigate the morphological changes in the hippocampus of newborn pups of rats subjected to experimental epilepsy during pregnancy. METHODS: Eighteen Swiss Albino rats were randomly divided into three groups (n=6): experimental group, saline-injected sham surgery group, and intact control group. In the experimental group of rats, an acute grand mal epileptic seizure was induced by 400 IU penicillin-G administration into their intra-hippocampal CA3 region with a stereotaxic device during the 13th day of their pregnancy. On the first neonatal day, pups were perfused with intracardiac fixative solution under anesthesia, and newborn hippocampi were dissected surgically for light and electron microscopic examinations. In an immunohistochemical study using Rat-401 mono-clonal antibody and peroxidase, nestin expression was analyzed in the developing hippocampal tissue. RESULTS: Histologically, normal migration and hippocampal maturation were determined in the newborn rat hippocampus in the control and the sham-operated groups. It was observed that the morphological structure of hippocampus in the experimental group corresponded to the early embryonal period. Most importantly, it was found that nestin (+)cell density was increased in the experimental epilepsy group in contrast to the control and sham groups. CONCLUSION: It has been concluded that epileptic seizures during embryonic life may cause impaired hippocampal neurogenesis and maturation,explaining the potentially harmful effects of epileptic seizures on the embryo at the early stage of neuronal differentiation. This is the first report regarding the alterations in nestin expression in newborn rat hippocampus. In the light of such findings, it will also be necessary to evaluate the functional consequences of a va-riety of epileptic seizures on learning and memory in neonates.

Differential effects of doxorubicin and docetaxel on nitric oxide production and inducible nitric oxide synthase expression in MCF-7 human breast cancer cells.

Anthracyclines and docetaxel are frequently used agents in the chemotherapy of breast cancer. In this study we examined the role of inducible nitric oxide synthase (iNOS) expression during the cytotoxicity of these drugs in the MCF-7 breast cancer cell line. Cell viability and cytotoxicity were evaluated by the trypan blue dye exclusion method. Apoptosis and necrosis were determined by the acridine orange/ethidium bromide dye method. The percentage of apoptotic cells was significantly higher with doxorubicin. However, total cytotoxic cell numbers were higher in the docetaxel group compared with doxorubicin, with respect to the control. Most of the cells were seen to be necrotic with the dye method. Cell extracts during the apoptotic process were applied to immunoblot by anti-iNOS monoclonal antibodies. While there was an increase in iNOS expression during docetaxel induced-cytotoxicity, a significant decrease in iNOS expression was detected during doxorubicin-induced cytotoxicity. The Griess method was used for detection of nitrate levels. It was compatible with immunoblot results. These data open a window for further studies to understand the mechanism underlining the cytotoxicity of docetaxel and doxorubicin.

Alterations of brain tissue in fetal rats exposed to nicotine in utero: possible involvement of nitric oxide and catecholamines.

Histopathological changes in the brains of embryos from female rats treated with nicotine during pregnancy and possible involvement of nitric oxide (NO) and catecholamines in the nicotine-induced abnormalities of developing brain were investigated. Sexually mature female Wistar rats were given 1, 2, and 3 mg/kg nicotine hydrogen tartrate (NHT) subcutaneously for 20 days after mating. Levels of cotinine, a nicotine metabolite, in the maternal plasma increased dose-dependently. Fetus and fetal brain weights were significantly lower in all nicotine-treated groups. Light microscopy of hippocampal CA1 area showed a decrease in the number of cells per unit area. Electron microscopy of the same region revealed a dose-dependent increase in intracytoplasmic edema, mitochondrial swelling, dilation of rough endoplasmic reticulum, nuclear configurative abnormalities, and condensation of the nuclear chromatin. Nitrate + nitrite levels in fetal brain homogenates were significantly lower in the groups treated with 2 and 3 mg/kg NHT. Norepinephrine and normetanephrine (NMN) levels were significantly higher in 2 and 3 mg/kg NHT groups, as well as dopamine, 3-methoxy-4-hydroxyphenylethyleneglycole (MHPG), and dihydroxyphenylacetic acid levels in the 3-mg/kg NHT group. In conclusion, maternal nicotine exposure may lead to structural abnormalities of the fetal brain tissue and may result in decreased levels of NO and increased levels of catecholamines and their metabolites.

The effect of experimental epilepsy induced by penicillin administration during pregnancy on nestin expression in the immature rat cerebellum. A light, electron microscopic, and immunohistochemical study.

INTRODUCTION: Recent knowledge regarding the effect of epileptic seizures in pregnant women on newborns was limited and, therefore, it was difficult to suggest the proper clinical guidelines and to take precautions against it. Studies evaluating the morphological effects of epileptic seizure during pregnancy on newborns in various experimental models are valuable. Therefore, the current study was designed to investigate the morphological changes in the cerebellum of newborn pups of rats subjected to experimental epilepsy during pregnancy. MATERIALS AND METHODS: Swiss Albino rats were divided into three groups (six animals in each). In the first group (experimental group) an acute grand mal epileptic seizure was induced by 400 IU penicillin-G administration into their intrahippocampal CA3 region with a stereotaxic device during the 13th day of their pregnancy. The second group (intrahippocampal saline-injected sham group) and the third group (untreated animals) were the control groups. On the 1st neonatal day, pups were perfused with intracardiac fixative solution under anesthesia, and newborn cerebellums were dissected surgically for light and electron microscopic studies. RESULTS: In an immunohistochemical study using Rat-401 monoclonal antibody and peroxidase, the intermediate filament nestin was detected in the developing cerebellar tissue. Histologically, normal migration and cerebellar maturation were determined in the newborn rat cerebellum in the control and sham-operated groups. It was observed that the morphological structure of the cerebellar cortex in the experimental group was compromised in the early embryonal period. In contrast to the control and sham groups, it was found that nestin (+) cell density was increased in the experimental epilepsy group. CONCLUSIONS: It has been concluded that epileptic convulsions during embryonic life may cause early neurogenesis and delayed maturation, which explains the harmful effects of epileptic grand mal seizures, hypoxia, and obstetric trauma to the embryo at the early stage of neuronal differentiation. However, further studies are necessary to investigate epileptic pregnant phenomena and to characterize the possible relationship between epilepsy and congenital malformations as well as mental retardation.

Changes in vascularity of cartilage endplate of degenerated intervertebral discs in response to melatonin administration in rats.

We carried out an experimental investigation of cartilage endplate vascularity of degenerated intervertebral discs produced by exogenous melatonin (MEL) treatment. Adult Swiss albino rats were divided into three groups: control, operated degeneration, and MEL treatment. There were five rats in each group and, using a posterior approach, cuts were made parallel to the endplates in the posterior annulus fibrosus in five consecutive intervertebral discs between the 5th and 10th vertebral segments of the rats' tails. At 8 weeks, five of these animals were treated with exogenous MEL (s.c. injection of 30 microg/100 g body weight daily for 4 weeks). In each experimental group, one animal was examined using CT scanner to study the density of the cartilage endplate of the disc. To evaluate the bone growth and vascularity of the cartilage endplate region, the animals were killed for subsequent histopathological evaluation. We found that the vascular channel counts and percentage areas from animals treated with MEL were significantly lower than from the operated degeneration animals. Accordingly, the density histogram in the MEL group showed a spike profile for both the vertebral body and the cartilage endplate, indicating an increase in the amount of higher density tissues in these regions. Our results demonstrate that the use of MEL reduces the cartilage endplate vascularity of degenerated intervertebral discs, suggesting that it may have an osteoinductive effect on bone formation. Further studies are needed to characterize fully the relevance of our findings for the treatment of disorders such as postmenopausal osteoporosis.

The effects of pineal gland transplantation on the production of spinal deformity and serum melatonin level following pinealectomy in the chicken.

Pinealectomy frequently produces spinal deformity in some animal models, but the precise biological mechanism of this phenomenon remains obscure. The current study investigated the effects of an autograft pineal body on the development of spinal deformity and serum melatonin (MLT) concentration after pinealectomy in the chicken. Thirty-six chickens (2 days of age) were divided into three equal groups. While the removal of the pineal gland was performed in groups B and C, a pineal body autograft was surgically implanted into the body wall musculature only in the pineal transplantation group (group C). Chickens in which no surgical intervention was performed served as intact controls (group A). Posteroanterior radiographs of the spines of the chickens were taken at the age of 8 weeks. These were used to determine Cobb angles and to measure the rib-vertebra angles (RVA) on the concave and convex sides of the curves, from which data the difference between the convex and concave RVA (the RVAD) was calculated. At the end of the study, serum MLT levels were determined using the enzyme-linked immunosorbent assay method, and histopathological examination of specimens from all the groups was performed. The results were compared using one-way analysis of variance followed by Duncan's test for pairwise comparisons or by the Kruskal-Wallis test followed by the Mann-Whitney U tests for comparisons between two groups. In this study, the serum MLT levels in groups B and C were significantly lower than those in group A ( P<0.05). However, scoliosis developed in only 7 of 12 (58%) in group B and 6 of 12 (50%) in group C. The average Cobb angle and RVAD in groups B and C were significantly larger than those found in group A ( P=0.000 and P=0.001, respectively). Interestingly, there were no significant differences in either serum MLT levels or development of scoliosis between groups B and C. From the results of the current study, it is evident that the intramuscular pineal gland transplantation following pinealectomy in young Hybro Broiler chickens has no significant effect on the development of spinal deformity and serum MLT level. In the light of this result, the role of MLT in the development of spinal deformity in chickens after pinealectomy remains controversial, and further investigations are warranted.

The effects of calcium channel antagonist nimodipine on end-plate vascularity of the degenerated intervertebral disc in rats.

The vascular channels at the end-plate of the intervertebral disc are very important in maintaining a healthy disc. With age, a reduction of the nutrition of the avascular nucleus pulposus is inevitable. On the other hand the calcium channel antagonist nimodipine has been shown to have a positive effect on blood flow in the region of the vertebral end-plate. To evaluate the effects of nimodipine on the end-plate vascularity in the degenerative discs, we have produced an experimental disc degeneration and evaluated the radiological and histopathological features of the end-plate of the degenerated discs. Adult rats were divided into 3 groups: control (n=5), operated degeneration (n=5), and nimodipine treatment (n=5). Using a posterior approach, a cut was made parallel to the end-plates in the posterior annulus fibrosus in 5 consecutive intervertebral discs between the 5th and 10th vertebral segments of the tails of adult Swiss Albino rats. At 8 weeks, 5 of these animals were treated with nimodipine. In each experimental group, 1 animal was examined using computed tomography (CT) to study the density of the cartilage end-plate of the disc. Then, the animals were sacrificed for subsequent histopathological evaluation. We found that the vascular channel counts and percentage areas from animals treated with nimodipine were higher than from both the non-operative control and operated degeneration groups, although these were not statistically different. Accordingly, the profile of the density histogram in the nimodipine-treated group showed a wide plateau, indicating an increase in the vascularity in this region. From our results, we suggest that nimodipine enhances vascularisation of the cartilage end-plate in the disc. It is possible that the increased proportion of vascular contacts at the end-plate has a beneficial effect in the nutrition of the disc. However, further experimental studies will be needed to determine the validity of this statement in animals or human beings.