Investigation of Demyelination and Remyelination Processes of the Compressed Optic Chiasm: An Experimental Model.

AIM: To demonstrated demyelination and remyelination of the optic nerve histologically by electron microscopy in an experimental model similar to the compression of pituitary adenomas on the optic chiasm. MATERIAL AND METHODS: The rats were fixed to a stereotaxic device under deep anesthesia, and a balloon catheter was placed under the optic chiasm through a burr hole which was in front of the bregma in accordance with the brain atlas of rats. The animals were divided into five groups (n=8): control, mild compression demyelination, severe compression demyelination, mild compression remyelination, severe compression remyelination. The fine structures of the tissues obtained were evaluated using electron microscopy. RESULTS: We found a significant difference in the severity of degeneration when comparing group 1 with group 5 (p < 0.001); there was no degeneration in group 1 rats and severe degeneration in all of the group 5 rats. Oligodendrocytes were found in all rats in group 1 and none of the rats in no group 2. The nuclei were preserved in the group 1 rats but damaged in all of the group 5 rats. There were no lymphocytes or erythrocytes in group 1 and all positives in group 5. CONCLUSION: This technique, which induced degeneration without causing damage to the optic nerve with toxic or chemical agents, revealed Wallerian degeneration similar to tumoral compression. After compression relief, the optic nerve remyelination process can be better understood, particularly for sellar lesions. In our opinion, this model may guide future experiments to identify protocols to induce and accelerate remyelination.

Anatomic and histologic features of diaphragma sellae that effects the suprasellar extension.

This study aimed to determine the anatomical and histological features of diaphragma sellae that affect the suprasellar extension of intrasellar tumours. Twenty-four fresh adult cadavers were dissected for the study. Diaphragma sellae and pituitary capsules with sellar structures were resected. The diaphragma sellae was anatomically reviewed in detail. Immunohistochemical staining was performed for collagen types I, II, III, and IV. We examined the suprasellar growth of 13 sellar tumours extending superiorly through the diaphragma sellae by performing a series of 2704 endoscopic transnasal operations to analyse the anatomic and histologic results of the study. The diameter of the foramen of diaphragma sellae varied between specimens. Of 24 specimens, the diaphragma sellae in five (21%) had a tight-type foramen and those in 19 (79%) were more spacious. An increased expression of collagen types I and IV was observed in the pituitary capsule and the diaphragma sellae. In this clinical series, we observed that all types of sellar tumours could expand through the foramen. We observed radiologically and intraoperatively that the diaphragma sellae was displaced laterally and formed a dome in two cases with an adenoma extending to the suprasellar area. Two types of suprasellar extension through the diaphragma sellae are possible: 1) The collagen structure of diaphragma sellae can be destroyed by invasive tumours; 2) The morphology of the foramen of the diaphragma sellae facilitates suprasellar tumoural extension. All sellar tumours, including non-invasive cystic tumours, may invade the suprasellar area by expanding through the foramen of the diaphragma sellae.

Effect of resveratrol and metformin on ovarian reserve and ultrastructure in PCOS: an experimental study.

BACKGROUND: PCOS is a reproductive hormonal abnormality and a metabolic disorder. It is frequently associated with insulin resistance, hyperandrogenism, chronic inflammation, and oxidative stress. We aim to investigate the potential therapeutic effects of combined therapy of resveratrol and metformin on polycystic ovaries via SIRT1 and AMPK activation. METHODS: Wistar albino rats were divided into control and experimental (PCOS) groups. DHEA-induced PCOS rats were given resveratrol (20 mg/kg/day), metformin (300 mg/kg/day) and combined therapy. At the end of the experiment, the body and ovarian weight of rats were measured and blood samples were analyzed for FSH, LH, testosterone, AMH, TNF-α and MDA levels. Histopathological evaluation of ovaries were carried out by light and electron microscopy. SIRT1 and AMPK immunreactivity and TUNEL assay were scored. Data were statistically analyzed by SPSS programme. RESULTS: Metformin and combined treatment groups reduced the body and ovary weights compared to the PCOS group. Serum testosterone levels were significantly higher in the PCOS group than in the control group and this was reduced when PCOS was treated with all but especially resveratrol. All the treatment groups decreased LH, LH/FSH, TNF-α and tissue AMH levels which were induced in the PCOS group, whereas metformin was unable to improve the increased MDA and plasma AMH levels. Treatment with resveratrol and/or metformin ameliorated the elevated number of secondary and atretic follicles and the decreased number of Graafian follicles in the PCOS group, which indicates the effect of the treatments on the maintenance of folliculogenesis. Light and electron microscopic findings supported the analysis of follicular count. Increased number of TUNEL (+) granulosa cells in the PCOS group were reduced significantly in the treatment groups. Resveratrol and metformin increased SIRT1 and AMPK immunreactivity, respectively, compared to the PCOS group. CONCLUSIONS: The results suggest that combined therapy of metformin and resveratrol may improve the weight gain, hormone profile and ovarian follicular cell architecture by inducing antioxidant and antiinflammatory systems via SIRT1 and AMPK activation in PCOS.

Morinda citrifolia L. (noni) and memantine attenuate periventricular tissue injury of the fourth ventricle in hydrocephalic rabbits.

This study was designed to evaluate the neuroprotective effects of Morinda citrifolia L. (Rubiaceae), commonly known as noni, and memantine (a N-methy-D-aspartate receptor inhibitor) on hydrocephalus-induced neurodegenerative disorders. Kaolin was injected into the cistern magna of male adult New Zealand rabbits to establish a hydrocephalus animal model. Memantine (20 mg/kg, intraperitoneally; memantine-treated group) or noni (5 mL/kg, intragastrically; noni-treated group) was administered daily for 2 weeks. Microtubule-associated protein-2 and caspase-3 immunohistochemistry were performed to detect neuronal degeneration and apoptosis in the periventricular tissue of the fourth ventricle of rabbits. Microtubule-associated protein-2 staining density was significantly decreased in the hydrocephalic group, while the staining density was significantly increased in the memantine- and noni-treated groups, especially in the noni-treated group. Noni treatment decreased the number of caspase-3-positive cells in rabbits with hydrocephalus, while memantine had no effect. These findings suggest that noni exhibits more obvious inhibitory effects on hydrocephalus-induced neurodegenerative disorders than memantine in periventricular tissue of the fourth ventricle.

Microsurgical anatomy of membranous layers of the pituitary gland and the expression of extracellular matrix collagenous proteins.

BACKGROUND: There are several reports about the microanatomical and histological features of sellar and parasellar membranous structures and clinical studies about MMP proteinase as a predictive factor. However, studies on collagen contents of sellar and parasellar membranous structures are limited. We demonstrated the membranous structures surrounding the pituitary gland and defined extracellular matrix (ECM) collagenous proteins, collagen I-IV expression patterns of sellar and parasellar connective tissues. METHODS: The study was carried out on ten fresh postmortem human bodies at the Forensic Medicine Institution. Cavernous sinuses were resected with sellar structures and were stored at -80°C liquid nitrogen tanks. Medial wall of the cavernous sinus, pituitary capsule and pituitary tissue samples were obtained for RT-PCR. Opposite side specimens were used for histological and immune staining studies. Collagens I-IV were studied by immunohistochemical and reverse transcription polymerase chain reaction (RT-PCR) methods. FINDINGS: The pituitary capsule and medial wall were identified as two different structures. The fibrous membrane, as the third membrane, was identified as staying whole in eight of ten specimens. Increased type IV collagen was determined in the pituitary gland, medial wall and pituitary capsule, respectively, in both RT-PCR and immunhistochemical studies. Immunhistochemical studies revealed that collagen I was strongly expressed in both the medial wall and pituitary gland. CONCLUSION: Increased type IV collagen was detected especially in pituitary tissue, the medial wall and the pituitary capsule by immune staining and RT-PCR. Type IV collagen was considered to be an important factor in the progression of adenoma and invasion.

Comparison of bipolar electrosurgical devices with ligatures and endoclips in the rat appendicitis model.

PURPOSE: The aim of this study is to compare bipolar radiofrequency-driven vessel sealer, bipolar electrocautery, polyglactin 910 sutures, and endoclips in appendiceal stump closure with respect to operative time, appendiceal stump strength, and inflammation in a rat appendiceal model. METHODS: Forty-eight Wistar-Albino rats, which had previously created appendicitis, were divided into 2 (groups A and B). Each group was further subdivided into 4 subgroups (AL, ABPC, AC, AS, BL, BBPC, BC, and BS) each containing 6 rats. An appendectomy was performed, and the stump was closed by bipolar radiofrequency-driven vessel sealer in the L, bipolar electrocautery in the BPC, endoclips in the C, and ligatures in the S subgroups. Cecum bursting pressures were determined instantly after the operation in group A and on the seventh postoperative day in group B. All operative times were measured. Appendices and appendiceal stumps underwent histopathologic examination. Statistical analyses were performed with Kruskal-Wallis and Mann-Whitney U tests. P < .05 was considered significant. RESULTS: Bursting pressures of the subgroups were comparable on postoperative immediate period and day 7. Operative times were significantly shorter in the L and BPC subgroups in A and B. Histopathologic examination showed that the inflammation scores were similar in group A. In group B, inflammation parameters were also similar except the necrosis status, which was found to be decreased in BL compared with BC and BS. Necrosis status was significantly lower, and lymphocyte quantity was significantly higher in BL and BBPC compared with AL and ABPC. CONCLUSIONS: Bipolar radiofrequency-driven vessel sealer and bipolar electrocautery achieve safe stump closure with satisfactory bursting pressure values in an experimental rat appendicitis model. Decreased operative time and unimpaired healing are encouraging.

Immunohistochemical analysis of TIMP-2 and collagen types I and IV in experimental spinal cord ischemia-reperfusion injury in rats.

BACKGROUND: Thoracic and thoracoabdominal aortic intervention carries a significant risk of spinal cord ischemia. The pathophysiologic mechanisms that cause hypoxic/ischemic injury to the spinal cord have not been totally explained. In normal spinal cord, neurons and glial cells do not express type IV collagen. Type IV collagen produced by reactive astrocytes is reported to participate in glial scar formation. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors that regulate the activity of the matrix metalloproteinases (MMPs). TIMP-2 binds strongly with MMP-2, facilitating activation by membrane-type MMP. Imbalance between TIMPs and MMPs can lead to excessive degradation of matrix components. Type IV collagen involved in the blood-brain barrier disruption and glial scar formation, TIMP-2 influences MMP-2 that controls degradation of collagen I and IV. OBJECTIVE: To examine the immunohistochemical analysis of TIMP-2 and collagen types I-IV in experimental spinal cord ischemia-reperfusion in rats. METHODS: Thirty-two male Wistar rats weighing 250-300 g were divided into four groups: group S: sham group (n = 8); group 0P: 30-minute occlusion without perfusion (n = 8); group 3P: 30-minute occlusion and 3-hour perfusion (n = 8); and group 24P: 30-minute occlusion and 24-hour perfusion (n = 8). Infrarenal aorta was cross-clamped at two sites by using two aneurysm clips for 30 minutes. Reperfusion was provided after removal of the clips. Lumbar spinal cord segments were removed for immunohistochemical analysis. RESULTS: TIMP-2 and collagen staining in 3-hour perfused (3P) group were nearly the same with sham group (S). TIMP-2 and collagen staining increased in the 24-hour perfused group. CONCLUSION: Alterations in collagen levels may relate to the biphasic breakdown of the blood-brain barrier and collagen staining in new cell types with relation to glial scar formation. Our results demonstrate that 3-hour perfusion after occlusion in hypoxic/ischemic spinal cord injury seems to be the critical reversible period.

LigaSure compared with ligatures and endoclips in experimental appendectomy: how safe is it?

PURPOSE: The present study aims to compare strength, healing, and inflammation of appendiceal stumps closed by LigaSure Precise (Valleylab, Boulder, CO, USA) device, ligatures using polyglactin 910 (Vicryl, Ethicon, Edinburgh, UK) and endoclips (Ligaclip ERCA, Ethicon, OH, USA), and operation time (OT) in experimental appendectomy. METHODS: Forty-eight Sprague-Dawley rats were divided into two (Group A and B). Each group was further subdivided into three subgroups (AS, AC, AL, BS, BC, BL) containing eight rats. Appendectomy was performed and stump was closed by ligatures in S, by endoclips in C and by LigaSure in L subgroups. OT was recorded. In Group A, cecum bursting pressures (BP) were determined instantly after the operation. In Group B, BP, histological evaluations, and measurements of collagen contents estimated by the tissue hydroxyproline (HPL) level were made on the seventh postoperative day. Statistical analyses were performed with Kruskal-Wallis test and Mann-Whitney U test. P value was considered significant at less than 0.05. RESULTS: BPs of subgroups were comparable on postoperative immediate period and day 7. HPLs and OTs were significantly better in L subgroups. BL had the least inflammation. CONCLUSION: Better healing, less inflammation, shorter OT, and equal strength achieved with LigaSure device comparing with polyglactin 910 ties and endoclips in experimental appendectomy is encouraging.

Biochemical effects of experimental epidural hematoma on brain parenchyma of rats.

INTRODUCTION: The management of epidural hematoma is classified into surgical or conservative treatment according to clinical and radiologic parameters. In the recent years, the number of paper suggesting conservative management has been increasing. The experimental works that have been performed are based on especially the effects of epidural hematomas. Basic pathophysiologic factors on ischemia result of brain trauma are based on biochemical mediators. Nitric oxide (NO) and malondialdehyde (MDA) are the substances that play important roles in brain damage after trauma. MATERIAL AND METHOD: In this study, 36 rats are divided into three groups (n = 12/group). Epidural hematoma was achieved by 0.1 ml autolog blood in rat epidural space with balloon model. Early and late phase biochemical effects on parenchyma of epidural hematoma operated in a volume which neither alters intracranial pressure (ICP) nor creates shift effect were observed. Biochemical changes of NO and MDA levels were examined in each of three experimental groups. RESULTS: NO values increased significantly in the early group (6 hours) compared with those in the control group. Difference of NO values between the control and late groups was not significant. An increase has been found in MDA values in the control group compared with those in the early group. MDA values of the late group (30 days) were closer to that of the control group. CONCLUSION: In this study, considering biochemical results, we have found that conservative volumes which neither increase ICP nor cause brain shift do not lead to permanent changes on brain.

Double myelomeningocele in a neonate: Case report and review of the literature.

The presence of myelomeningocele at multiple levels along the spinal column is extremely rare. There are only a few cases of double myelomeningocele reported in the literature. We report the case of a newborn male who had two myelomeningoceles with one at the cervical level and the other at the lumbar level. This unusual case report as well as a review of the literature is presented.

Localization of pan-cadherin immunoreactivity in adult rat tissues.

Cadherins, being responsible for selective cell recognition and normal tissue integrity in adults, regulate morphogenesis in a variety of organs during development. In this study, anti-rat pan-cadherin antibody, specific to all subgroups of the cadherin family, was used to map the distribution of the pan-cadherin immunoreactivity in adult rat organs. Pan-cadherin immunoreactivity positive tissues were: secretory cells of the adenohypophysis, autonomic nerve, corneal epithelium, oesophageal nerve plexus, stomach and pyloric glandular cells, epithelium of the ileum and its nerve plexus, alveolar cells of the lung, proximal convoluted tubules of the kidney, islet cells of Langerhans, and the acinar cells of the exocrine pancreas. For the first time, positive pan-cadherin immunoreactivity was demonstrated in the epithelial cells of the corpus ciliaris and in the nerve plexus of corpus cavernosum of the penis. In conclusion, our results suggest that cells in many tissues and organs of the adult rat synthesize cadherins.