We investigated the effects of a 900 Megahertz (MHz) electromagnetic field (EMF), applied during the prenatal period, on the spleen and thymus of 21-day-old male rat pups. Pregnant Sprague-Dawley rats were divided into control and EMF groups. We applied 900 MHz EMF for 1 h/day to the EMF group of pregnant rats. Newborn male rat pups were removed from their mothers and sacrificed on postnatal day 21. Spleen and thymus tissues were excised and examined. Compared to the control group, thymus tissue malondialdehyde levels were significantly higher in the group exposed to EMF, while glutathione levels were significantly decreased. Increased malondialdehyde and glutathione levels were observed in splenic tissue of rats exposed to EMF, while a significant decrease occurred in superoxide dismutase values compared to controls. Transmission electron microscopy showed pathological changes in cell morphology in the thymic and splenic tissues of newborn rats exposed to EMF. Exposure to 900 MHz EMF during the prenatal period can cause pathological and biochemical changes that may compromise the development of the male rat thymus and spleen.

Mercury is ubiquitous in the environment; it is an occupational pollutant and a potential toxicant. We investigated the effects of exposure of rat testes to mercury vapor (Hg(0)). Twelve male rats were divided into two groups of six: the rats of the Hg(0) group were exposed to mercury (1 mg/m(3)/day) in a chamber for six weeks; the control group rats were housed under the same conditions without exposure to Hg(0). After the experimental period, the testes were removed, sections of testis were evaluated histopathologically after hematoxylin and eosin staining, and stereologically using the Cavalieri principle and optical fractionator methods. We found significant decreases in the total volume of testis, diameters of seminiferous tubules and total volume of seminiferous tubules. Significant decreases were detected in the numbers of Sertoli cells, spermatogonia, spermatocytes and spermatids of the Hg(0) group compared to the control group. In the Hg(0) exposed group, spermatogenic cells were degenerated and seminiferous tubules were atrophied.

Oxidative stress is one of the main reasons of both menopause and diabetes. So, it plays crucial role in the pathogeneses of that condition and disease. Therefore, the objective of the present study was to investigate the effects of menopause and diabetes upon the hippocampus using a rat model. Adult female Sprague Dawley rats (n = 24) were allocated randomly as follows; control (C group) ovariectomized (O group), diabetic (D group) and ovariectomy plus diabetic groups (DO group) (n = 6; in each group), respectively. For evaluating the results, tissue biochemistry and stereological analysis were made. Biochemistry results (lipid peroxidase (LPO); catalase (CAT); superoxide dismutase (SOD); total glutatyon (GSH); and myeloperoxidase (MPO) values) in Group C-DO were determined as 12.27, 21.88, 23.08 and 29.90 nmol/gr tissue; 59.3, 70.06, 69.7 and 78.1 mmol/min/mg tissue; 174.2, 156.4, 159.7 and 154.6 mmol/min/mg tissue; 3.63, 3.61, 4.21 and 3.97 nmol/mg tissue; and 5.05, 5.68, 5.58 and 6.19 µmol/min/mg tissue, respectively. Moreover, both menopause and diabetes led to change of lipid profiles. There were significant differences between the control and other groups (Group C and D-DO) (p < 0.01) and among experimental groups (p < 0.01) in terms of neuron number. When the volumes of the hippocampus were compared, there were no significant differences between the all groups (P > 0.05). At this point, we suggested that diabetes could aggravate deleterious effects of ovariectomy.

We studied with quantitative and microscopical methods the heart of rats divided into five age groups: embryos at the age of 11 days, fetuses at the age of 16 days and 20 days and also heart samples of 3-day-old pups and young adults (5 weeks of age) were used (n = 10 samples in each group). At the end of the study; heart samples were obtained from all animals. Stereological estimations were performed on heart volume, volume of heart lumen (ventricles and atria), volume of myocardium, numerical density of the myocyte nuclei and mean nuclear diameter of myocytes. Volumetric values and numerical data were estimated via Cavalieri method and physical dissector, respectively. In this study, histological examination was performed at light and electron microscopic levels. The numerical density of the myocyte nuclei increased from fetuses to young adults. Differences between embryos and fetuses, between fetuses and 3-day-old pups, and between 3-day-old pups and young adults were statistically significant. These results indicate that myogenesis continued in the rat myocardium during prenatal life and after birth.

BACKGROUND:Diabetes mellitus (DM) has a negative effect on cardiovascular functions. Little, however, is known of the overall effect of DM on the cardiac histology or the pathophysiological basis of this.AIM:We aimed to investigate the role of oxidative stress on the pathogenesis of diabetic cardiomyopathy in an experimental model.MATERIALS AND METHODS:12 week-old female Sprague Dawley rats were randomly allocated into a healthy control group (n=6) and an DM group (n=6). After 12 weeks of alloxan induced DM, the groups' cardiac tissues were histopathologically analyzed and examined for determination of oxidant and antioxidant enzymes [activities of catalase (CAT), superoxide dismutase (SOD), and myeloperoxidase (MPO) and amount of reduced glutathione (GSH) and lipid peroxidation (LPO)].RESULTS:When compared to the control group, the DM group showed cardiomyopathic changes. In the DM group, activities of CAT (144 +/- 0.9 vs. 112 +/- 1.4, p < 0.05) and LPO amount (27.0 +/- 0.74 vs. 14.4 +/- 0, 20, p < 0.05) were significantly increased whereas activities of SOD (142 +/- 0.2 vs. 146 +/- 0.7, p < 0.05) and amount of GSH (3.48 +/- 0.01 vs. 3.73 +/- 0.01, p < 0.05) were significantly decreased when compared to the control group. Besides, activities of MPO (7.3 +/- 0.02 vs. 8.6 +/- 0.11, p < 0.05) were comparable between groups.CONCLUSIONS:Using the experimental animal model, we were able to demonstrate that DM causes cardiomyopathic changes, and we propose that these changes could be mediated by an oxidative stress.

Sepsis is a systemic inflammatory response to infection and a major cause of morbidity and mortality. Sildenafil (SLD) is a selective and potent inhibitor of cyclic guanosine monophosphate (cGMP)-specific phosphodiesterase PDE5. We aimed to investigate the protective effects of sildenafil on caecal ligation and puncture (CLP)-induced sepsis in rats. Four groups of rats were used, each composed of 10 rats: (i) 10 mg/kg SLD-treated CLP group; (ii) 20 mg/kg SLD-treated CLP group; (iii) CLP group; and (iv) sham-operated control group. A CLP polymicrobial sepsis model was applied to the rats. All groups were killed 16 h later, and lung, kidney and blood samples were analysed histopathologically and biochemically. Sildenafil increased glutathione (GSH) and decreased the activation of myeloperoxidase (MPO) and of lipid peroxidase (LPO) and levels of superoxide dismutase (SOD) in the septic rats. We observed a significant decrease in LPO and MPO and a decrease in SOD activity in the sildenafil-treated CLP rats compared with the sham group. In addition, 20 mg/kg sildenafil treatment in the sham-operated rats improved the biochemical status of lungs and kidneys. Histopathological analysis revealed significant differences in inflammation scores between the sepsis group and the other groups, except the CLP + sildenafil 10 mg/kg group. The CLP + sildenafil 20 mg/kg group had the lowest inflammation score. Sildenafil treatment decreased the serum tumour necrosis factor (TNF)-α level when compared to the CLP group. Our results indicate that sildenafil is a highly protective agent in preventing lung and kidney damage caused by CLP-induced sepsis via maintenance of the oxidant-anti-oxidant status and decrease in the level of TNF-α.

The purpose of this study was to determine the amount of atrophy occurring in the cerebrum of patients with multiple sclerosis compared with controls, and to show the relationship between clinical status and distribution of atrophy. MR images were obtained on 12 relapsing-remitting (RR) and 11 secondary progressive (SP) multiple sclerosis patients and 24 control subjects (all patients and controls were female). The Cavalieri method by modern design stereology was used to measure the cerebral volume. It was found that volumes for RR and SP with multiple sclerosis and control subjects were 757242 mm(3), 716867 mm(3) and 912499 mm(3) respectively. Mean estimates of volume loss in RR and SP were 20.5% and 27.2% respectively compared to controls for the cerebrum. In addition the volume difference between RR and SP was 5.6%. There were statistically significant differences between both RR (P<0.05) and SP (P<0.05) compared with control subjects but no differences between RR and SP volumes. MRI-estimated cerebrum volumes may be help to evaluate patients' clinical status and provide a simple index to assess the efficiency of therapy.

Locating the same microscopic fields in consecutive sections is important in stereological analysis. The tools for achieving this requirement have limited number in practice. This paper presents a simple and inexpensive technique for localizing the same fields on disector pairs in conventional light microscopes equipped with widely available dial indicators. It is partly a modification of equipment previously described. The presented procedure requires two light microscopes equipped with dial indicators and modified slide clips. An application of the present system was shown in a model of spinal cord injury (SCI). A midthoracic laminectomy was performed leaving the dura intact. A contusion was done at the level of midthoracic spinal cord segments (T7–T8) by dropping a 10-g mass from a height of 30cm. The subjects were randomly divided into three groups (10 animals in each): hypothermia group, methylprednisolone group, and traumatic spinal cord injury alone group. Present results show that treatment with hypothermia after spinal cord trauma has a neuroprotective effect on cell damage but not in the methylprednisolone treatment group.

Zinc (Zn) is an essential trace element for humans and animals. It is required for normal growth, gene expression, wound healing, protein metabolism, immune function, and membrane integrity. In this study, unbiased stereological methods have been used to quantify the effects of Zn deficiency on the sectioned surface area and the number of myelinated axons in the sciatic nerve of rats. Animals were fed a Zn-deficient or Zn-sufficient diet for a period of 4 weeks. At the end of this time, the samples of sciatic nerves were removed from the animals, processed for electron microscopy and embedded in resin. The Zn-deficient group of rats was found to have a lower body weight compared to rats in the control group (P < 0.05). The sectioned surface area of nerve cross-section and myelinated axon number in Zn-deficient rats decreased by 20% and 29%, respectively, compared to the control group. A significant correlation between sectioned surface area and myelinated axon number was also determined. Morphological findings were as follows: on light microscopy, it was determined that certain abnormalities occur specifically in the experimental group, such as collapsed nerve fascicles, irregular profiles of and degeneration in myelin sheaths, and on electron microscopy, extensive myelin damage was seen in Zn-deficient groups compared with control groups. This study suggests that peripheral nerves require Zn for development and preservation of their structure.

in recent studies, some neuroleptics have shown neurotoxic activities. Clinical and experimental studies have been carried out to investigate the effects of different neuroleptic drugs considered to affect the central nervous system. The aim of this study was to evaluate neurotoxic effects of haloperidol on hippocampal neurons. The drug was given in daily doses of either 1 or 3 mg/kg for 6 weeks to adult male guinea pigs. After treatment, all animals were anaesthetized via short inhalation of ether, and then were fixed by a mixture of 2% glutaraldehyde and +2% paraformaldehyde in 0.1 M phosphate buffer. Brains were removed from the cranium and stored in the same fixative overnight. On the following day, the CA1 region of the hippocampus was dissected out. After embedding in araldite resin and obtaining semi-thin sections, the tissues were stained with toluidine blue. The physical disector was used for measurements of nuclear height and numerical density of neurons and the sections were also evaluated histopathologically. The numerical density of neurons and nuclear height in the hippocampus for the low-dose (1 mg/kg) and high-dose (3 mg/kg) experimental groups were 121476+/-13375 cell/mm(3) and 3.6 mum and 71113+/-8775 cell/mm(3) and 3.56 pm, respectively. In contrast, the control group had a neuronal numerical density of 165498 15990 cell/mm(3) and a nuclear height of 4.09 mum. There was a significant difference in both the mean density of neurons and the mean height of nuclei between haloperidol-treated and control groups (p<0.05). There was also a statistical difference in the mean density of neurons (but not in nuclear height) when comparing the dosage of haloperidol (p<0.05). These findings suggest that haloperidol treatment may lead to a loss of neurons as well as a decrease in the height of nuclei in the hippocampus.

In this study, the anti-inflammatory effect of erythromycin was investigated in a model of histamine-induced otitis media with effusion (OME). OME was induced in guinea pigs by the transtympanic injection of histamine solution into the middle-ear cavity. Guinea pigs were randomly assigned to one of three groups: control, erythromycin treatment, or methylprednisolone treatment. After histamine injection, the animals were treated with intraperitoneal medication for five days consecutively. Afterwards, the animals were sacrificed and the temporal bones were removed. The samples were examined stereologically. In the erythromycin-treated group, it was observed that neutrophil infiltration was significantly inhibited when compared to the control group. This result shows that erythromycin may produce a significant anti-inflammatory effect in this model of OME.

Oxcarbazepine (OXC) is a widely used novel antiepileptic drug that has been available for routine prescription for 10 years. To examine low dose OXC-induced neurotoxic effects on cerebellar development, we administered 25 mg/kg OXC orally to newbom Wistar rats once a day on postnatal days 2-14. Microscopic processed cerebellar sections of the control and treated groups were examined by volumetric analysis. Volume estimations were obtained using the Cavalieri's principle using a computerized stereological image analyzer (CAST-GRID). The total volume of the cerebellum, white matter and the various cerebellar layers (except extragranular layer) were significantly increased in the treated animals. These data may provide useful implications for the management of OXC-induced developmental neurotoxicity in children exposed to OXC during the late fetal period. Our finings suggest that women suffering from epilepsy should be given OXC carefully only at the lowest effective doses during pregnancy.

In recent studies, some neuroleptics have shown neurotoxic activities. Clinical and experimental studies have been carried out to investigate the effects of different neuroleptic drugs considered to affect the central nervous system. The aim of this study was to evaluate neurotoxic effects of haloperidol on hippocampal neurons. The drug was given in daily doses of either 1 or 3 mg/kg for 6 weeks to adult male guinea pigs. After treatment, all animals were anaesthetized via L short inhalation of ether, and then were fixed by a mixture of 2% glutaraldehyde and +2% paraformaldehyde in 0.1 M phosphate buffer. Brains were removed from the cranium and stored in the same fixative overnight. On the following day, the CAl region of the hippocampus was dissected out. After embedding in araldite resin and obtaining semi-thin sections, the tissues were stained with toluidine blue. The physical disector was used for measurements of nuclear height and numerical C density of neurons and the sections were also evaluated histopathologically. The numerical density of neurons and nuclear height in the hippocampus for the low-dose (1 mg/kg) and high-dose (3 mg/kg) experimental groups were 12.4 mm(3) and 3.6 mum and 7.14 mm(3) and 3.56 mum, respectively. In contrast, the control group had a neuronal numerical density of 16.55 mm(-3) and a nuclear height of 4.09 mum. There was a significant difference in both the mean density of neurons and the mean height of nuclei between haloperidol-treated and control groups (p<0.05). There was also a statistical difference in the mean density of neurons (but not in nuclear height) when comparing the dosage of haloperidol (p<0.05). These findings suggest that haloperidol treatment may lead to a loss of neurons L as well as a decrease in the height of nuclei in the hippocampus.

Haloperidol is a widely used antipsychotic drug, which exerts its effects via antagonizing the dopaminergic D2 receptors. Also it affects a number of receptors on vascular bed and other tissues. The impact of haloperidol on vascular bed seems still debatable and not clear. In the present study, haloperidol was given to adult rats in 0.5, 1, 2.5 and 5mgkg−1 doses, once a day, intraperitoneally in 1ml volumes, for 9 weeks. After decapitation under Pentothal anesthesia, brains and basilar arteries were dissected out at midpontine level immediately. Conventional histopathology and morphometric analysis were carried out on the dissected artery branches.Medial and adventitial layers, endothelial cells and internal elastic membranes were observed as normal in the control group. It was determined clearly that the lumen of basilar artery in the control group was larger than in the other groups and also it was observed that is more regular the lumen contours of basilar artery in control group compared with other groups. Finally, wall thickness of basilar artery in all experimental groups decreased significantly due to the vasoconstriction. Regarding the total, lumen and wall volumes, 1mgkg−1 haloperidol induces vasoconstriction more than the other groups.

ABSTRACTBackground Behçet's syndrome is a chronic systemic immuno‐inflammatory disorder affecting multiple organs with generalized vasculitis of arteries and veins. Although the aetiology is still unknown, endothelial dysfunction is one of the most prominent features in Behçet's syndrome. The skin pathergy reaction (SPR) is a non‐specific hyperreactive lesion formation that is one of the major features and diagnostic criteria of the disease. It develops after 24–48 h at the site of the needle‐prick, especially in the exacerbation period, and it is very similar to the erythematous papules or pustules that appear spontaneously in patients with Behçet's syndrome. Therefore, an investigation into the formation of the SPR lesion may contribute to the pathophysiology of skin lesions of this unique disorder.Objective To evaluate the immunological features of SPR formation by assessing the immunohistochemical staining of cell adhesion molecules and endothelial growth factor markers such as E‐selectin, P‐selectin and endoglin (CD 105).Methods Patients with Behçet's syndrome showing positive (n = 15) or negative (n = 10) SPR and 15 age‐ and sex‐matched hospital‐based healthy control subjects from a similar ethnic background were included in this study. Patients were divided into active and inactive stage by clinical findings and acute‐phase reactant parameters including erythrocyte sedimentation rate (ESR) and neutrophil count. Punch biopsy specimens were obtained both from the lesion site on the forearms at 48 h and from normal skin approximately 5 cm adjacent to the SPR site. A biopsy was also obtained from the test application site in Behçet's syndrome patients with negative SPR and healthy volunteers. Biopsy specimens were then evaluated by immunohistochemical staining.Results Immunohistochemical examination demonstrated a mixed inflammatory cell infiltrate around the vessels and skin appendages that extended somewhat into the deep dermis. A positive segmental staining of E‐selectin and P‐selectin was noted in the endothelial cells of biopsies obtained from the patients with positive SPR. A positive segmental staining of CD 105 in the endothelial cells was also observed in the same group of patients. However, the immunostaining of the same markers was found to be negative in the biopsies obtained from normal skin of SPR‐positive patients, SPR‐negative patients and healthy control subjects. Both acute‐phase reactant levels were significantly higher in the active stage than in inactive patients or healthy controls.Conclusion Interaction of cellular adhesion molecules together with endothelial proliferation may play an important role in the formation of SPR lesions in patients with Behçet's syndrome. The involvement of the vascular endothelium in a large number of diseases including Behçet's syndrome supports the importance of vascular‐specific adhesion molecules for their aetiopathogenesis.

Hippocampal complex of the avian brain is one of the most studied brain regions in recent years. This structure is thought to be homologous to mammalian hippocampus and have an important role in spatial memory. Connection patterns between the avian hippocampus and other brain regions strongly support the idea of homology between mammalian and avian hippocampal complexes.Total neuron number of chick hippocampus in different ages was estimated using optical fractionator counting method. Animals were separated into two groups as one day- and seven day-old chicks. The mean total number of nerve cells in the right hippocampi of one day-old and seven day-old chicks was estimated as 783 310 +/- 12 749 and 760 028 +/- 12 256 respectively. The difference between two groups was not statistically significant (p > 0.05).The mean total volume of hippocampi of chicks was estimated by Cavalieri's principle and it was found to be 2.48 +/- 0.08 mm(3) for one day-old chicks and 2.80 +/- 0.04 mm 3 for seven day-old chicks. The difference between these volume values was statistically significant (p < 0.01).Although the difference in total neuron numbers between two groups was not raised statistically significant level, decreasing cell number with age might be depend on the destruction-of unneeded nerve cells with aging. The increased volume in seven day-old group may be associated with enlargement due to several reasons during development.

Tumor of the follicular infundibulum (TFI) is a rare epithelial tumor, and its histogenesis is still somewhat debatable. The diagnosis of TFI cannot currently be made before biopsy, and it is most often misdiagnosed as basal cell carcinoma (BCC). A 78-year-old woman presented to the dermatology clinic with a slightly erythematous nodule surrounded by fine telangiectasia on her right temple. Histopathologic examination of the lesion revealed an epithelial tumor with a plate-like growth pattern as is typical of TFI. In the same specimen, an unusual trichilemmal tumor with multiple epidermal connections was observed adjacent to the foci of TFI. The lobular tumor was composed of a large mass of keratinocytes with pale cytoplasm arranged as sheets and interconnecting ribbons. Several cells with hyperchromatic nuclei and small foci of compact cornification were also noted within the bulk of the tumor. The histopathologic features of the unusual tumor were reminiscent of BCC with trichilemmal differentiation or desmoplastic trichilemmoma.

The effects of passive avoidance learning on synaptic morphology and number in the dorsolateral hippocampus of chick were investigated at 24 and 48 h after training. Chicks of both sexes were used. The numerical density of synapses and mean synaptic height were determined using design-based quantitative electron microscopic techniques. Our results suggest that after training there is a significant increase in synaptic density in the dorsolateral hippocampus of chicks at both 24 and 48 h, and also that the mean synaptic height was significantly different between trained and control groups. The increase in synaptic density was due to shaft (type II) synapses. It is known that during synaptogenesis, shaft synapses are formed first and are then converted to spine synapses. The only hemispheric asymmetry was found in the 24 h water-trained (W-trained) males where the numerical density of spine synapses was significantly higher in the left hippocampus. No significant differences due to gender in either numerical synaptic density or synapse height were observed at either 24 and 48 h. Comparison of the 24 h with 48 h groups showed an increase in shaft synaptic density over time in the W-trained groups, and an increased density of both shaft and spine synapses with time in methylanthranilate-trained (MeA-trained) chicks. These results demonstrate that the dorsolateral hippocampus of the chick shows synaptic changes at both 24 and 48 h after training and implicates this region in the long-term memory process.

BACKGROUND:Tamoxifen (Tx) is used mostly in the treatment of breast and gynecological cancers. It is also widely used in the treatment of different dermatological disorders. However, its effects on skin have not been investigated previously.OBJECTIVE:To investigate the effects of Tx administration on rat skin.METHODS:Forty Spraque-Dawley female newborn rats were separated into two control groups and two experimental groups (n 10). One day after birth, the control groups of newborn rats were given 0.02 ml saline subcutaneously (s.c.) daily whereas experimental litters were treated with 100 microg Tx citrate in 0.02 ml saline s.c. daily for five days. The first control group and experimental group of rats were anesthetized at 21 days whereas the second control group and experimental group of rats were anesthetized on the 28th day. Histopathological assessments were made and compared with the control groups.RESULTS:Abnormal hair follicles were observed in both experimental groups of rats. Epidermal atrophy together with increased dermal fibrosis was more prominent in the first experimental group. Dermal fibrosis and lymphohistiocytic inflammatory cell infiltration were found to be prominent around the hair follicles in the second experimental group.CONCLUSION:Considerable harmful effects of Tx administration were observed on rat skin.

A 3-year-old Turkish boy with a history of chronic cough, recurrent bronchopneumonia, and a borderline sweat chloride test (40 mEq/L) was referred for further evaluation to our department. He was born at term (2100 g) to a marriage with no consanguinity. His mother and father were 40 and 46 years old, respectively. Physical examination (Fig. 1) revealed hypopigmented, atrophic, and hyperkeratotic skin lesions surrounded by reticulate hyperpigmentation on the entire body, predominantly on the face, neck, arms, shoulders, and legs, which had been noticed initially at the age of 18 months. Dystrophic toenails, sparse and thin hair, and phimosis were also observed. Laboratory tests disclosed an isolated neutropenia (white blood cell count, 1800/mm3). Bone marrow (BM) aspiration showed a decreased myelopoiesis without myelodysplastic changes, but normal erythropoiesis, megakaryopoiesis, and normal stroma. Lymphocyte subgroups containing CD4, CD5, CD6, CD8, CD19, CD23, and CD25, and immunoglobulin G (IgG), IgM, IgA, and IgE, were in the normal range; hemoglobin F (HbF), 2.8%. Spontaneous and clastogen-induced chromosome breaks were not increased. A skin biopsy showed increased pigmentation at the basal layer, dyskeratotic epidermal cells, and marked IgM deposition and cytoid bodies and mild IgA and IgG deposits at the dermo-epidermal junction. Lactate response to glucose challenge, amino acid chromatography, and urine organic acid analysis were normal. A diagnosis of dyskeratosis congenita (DC) was made with typical skin lesions, dystrophic toenails, thin and sparse hair, and neutropenia with decreased myelopoiesis in BM. Treatment with granulocyte colony-stimulating factor (G-CSF) was considered for the neutropenia. As the increase in neutrophil count at a dose of 5 microg/kg was not adequate, 10 microg/kg G-CSF was tried (Fig. 2). With 10 microg/kg once to three times a week, a 1.8-4.8-fold increase in the absolute neutrophil count (ANC) was achieved with no side-effects. Treatment was more frequent during infection (days 22-28).