In this study, we evaluated the protective effects of combined melatonin and vitamin D3 treatment on ovarian reserve and tissue architecture in a cyclophosphamide-induced premature ovarian failure (POF) rat model. Forty-nine adult female rats were randomly assigned to seven groups, including intact control (group 1), single-agent control (groups 2 and 3), POF (group 4), and POF + treatment (groups 5, 6, and 7) groups. Cyclophosphamide exposure led to elevated FSH and LH levels, reduced estradiol and progesterone levels, extensive follicular atresia, stromal fibrosis, and the marked degeneration of the ovarian ultrastructure. Additionally, the expression levels of PTEN, FOXO3a, and AMH were significantly downregulated, while caspase-3 and TNF-α immunoreactivities were increased. Notably, co-treatment with melatonin and vitamin D3 preserved primordial and growing follicle populations, restored hormonal balance, reduced stromal fibrosis, and attenuated apoptosis and inflammation markers. These findings highlight the potential of combined melatonin and vitamin D3 therapy as a fertility-preserving strategy that functions by mitigating chemotherapy-induced ovarian injury through multi-pathway modulation.

In this study, we evaluated the effects of anti-Müllerian hormone on follicle development and oocyte quality with light and electron microscopy. Twenty-four adult female rats were divided into four groups. After estrous cycle synchronization, on the first day, control group rats were injected with 0.5 ml saline, 2nd, 3rd, and 4th groups were injected 1 µgr, 2 µgr, and 5 µgr anti-Müllerian hormone, respectively. On the third day, intracardiac blood samples were taken for follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone serum level measurements. Ovaries were obtained for light and electron microscopic examinations. Secondary (antral) follicles were decreased while atretic follicles were increased in number parallel with an increased dose of anti-Müllerian hormone injection. Atresia of the follicles was demonstrated with apoptosis of granulosa cells characterized by apoptotic bodies and with paraptosis characterized by the vacuole formation in the cytoplasm, enlargement of granular endoplasmic reticulum cisternae and perinuclear cisternae in granulosa cells. Premature luteinization characterized by increased lipid droplets, mitochondria with tubular cristae, and smooth-surfaced endoplasmic reticulum in the cytoplasm of granulosa cells were detected in some growing follicles. In the anti-Müllerian hormone injected experimental groups, cystic follicles characterized by a large antrum, attenuated granulosa cell layer, and flattened granulosa cells that face the antrum were observed. Corpus luteum and stroma were similar in all groups. It was concluded that increasing doses of anti-Müllerian hormone caused increased atresia in developing follicles, premature luteinization of granulosa cells in some follicles, and cystic follicle formation in the further developing follicles.

Histoloji, hücre, doku ve organların normal yapılarını mikroskop altında inceleyen bilim alanıdır. Histoloji eğitiminde mikroskobik incelemelerin önemi büyüktür. Tıp eğitiminde Dönem I ve Dönem II ders programlarında verilmekte olan Histoloji ve Embriyolojiprogramı, teorik dersler yanında laboratuvar uygulamalarını da içermektedir. Hücre, doku ve organların mikroskop altındaki görüntülerinin kavranması ve öğrenilmesinde, histoloji atlasları yanında, şekillerin bizzat öğrenciler tarafından çizilmesi kalıcılığı artırmaktadır. Histoloji Laboratuvar eğitiminde uzun yıllardan buyana kullanılan Histoloji Laboratuvar Kılavuzu her yıl yeniden gözden geçirilerek öğrencilerin eğitimine sunulmaktadır.

In this study, we evaluated the structural features of the endometrial tissues, the immunohistochemical expression of MUC-1, which plays an important role in implantation, and the biochemical markers during the implantation window. Randomly chosen 18 fertile and 18 unexplained infertile women that have 27-32 days long menstrual cycle, normal hormonal values, normal USG findings of ovary and endometrium were included. Five, six, and seven days after ovulation, endometrial biopsies were taken and prepared in accordance with light and electron microscopy tissue preparation methods. Immunohistochemical methods were used to determine MUC-1 expression in the tissues. Serum hormone levels were determined. The MUC-1 immunoreactivity, as well as the serum levels of FSH, LH, TSH, estrogen, progesterone, and total testosterone did not differ significantly between the two groups; however, prolactin levels were higher in the infertile group. In the unexplained infertile samples, intraepithelial lymphocytes were frequently observed, the microvilli of the surface columnar epithelium were widespread, cells with pinopodes as well as vesiculated cells were minimal, pinopode development was insufficient, and the development of the endometrial glands was deficient. It was concluded that these structural differences observed in the surface and glandular epithelium of the endometrium in unexplained infertile patients may be due to the insufficiency of these cells in responding to steroid hormones; therefore, these changes may affect the implantation of the blastocyst in the endometrium.

The aim of this study was to demonstrate the effects of vitamin D treatment on ultrastructural changes and AMHR2 expression in the ovary in PCOS rat model. A total of 24 female prepubertal rats were divided into 3 groups. In group 1, sesame oil was injected and used as control group. In group 2, PCOS was created by the injection of 6 mg/kg/day DHEA. In group 3, PCOS was created and 120 ng/100 g 1,25 (OH)(2)D3 treatment was performed. At the end of the 28th day, the blood samples were collected. The ovarian tissues were obtained for electron microscopic and immunohistochemical examinations. Serum AMH, testosterone, FSH, LH levels and LH/FSH ratios were higher in the PCOS group compared to the control group and decreased in the treatment group compared to the PCOS group. AMHR2 expression was increased in atretic and premature luteinizate antral follicles in the PCOS group compared to the control group, and decreased in the treatment group compared to the PCOS group. PCOS group electron micrographs showed degenerative changes in developing follicles, cystic follicles characterised with granulosa cell layer attenuation and thickening of the theca cell layer, and lipid accumulation in the interstitial cells. Structural changes observed in the PCOS group were improved with vitamin D treatment. As a result, there is an interaction between PCOS, AMH serum levels and AMHR2 in the ovarian follicles. Vitamin D has a positive effect on hormonal and structural changes in the PCOS group. We concluded that vitamin D supplementation may be beneficial in PCOS patients.

In this study, we aimed to investigate the structural changes seen in the endometrium in experimental PCOS rat model and the effects of vitamin D treatment on these changes at immunohistochemical and electron microscopic levels. 24 prepubertal female rats were divided into three groups. Two groups were injected with dehydroepiandrosterone and one of them was treated with 1,25(OH)2 D3 at the same time. The control group was injected with sesame oil. At the end of the 28th day, the blood samples were collected. Uterus tissues were prepared for light and electron microscopic examinations. Epithelial, stromal and endometrial thickness measurements were investigated. Immunohistochemical staining was applied against caspase-3 and Ki-67. Serum AMH and estradiol levels were higher in PCOS group compared to the control group. Serum progesterone levels were similar in all groups. Endometrial, epithelial and stromal thickness measurements were increased in PCOS group compared to the control group, and decreased in the vitamin D treatment group compared to the PCOS group. Light and electron microscopic results of PCOS group showed an increase in apoptosis and proliferation. In the PCOS group, immunohistochemical staining of caspase-3 and Ki-67 were found to be higher than in the control group, but stainings were decreased with vitamin D treatment compared to PCOS group. Structural changes observed in endometrium may be related to implantation problems seen in patients with PCOS. Our studies suggest that vitamin D therapy may be beneficial in these patients.

OBJECTIVE:In our study, we aimed to investigate histopathological effects of chronic methyl parathion exposure on ovaries at electron microscopic level.MATERIALS AND METHODS:In this study, Wistar albino type, adult, female rats with an average weight of 190-250 g were used. 30 female rats, included in this study, were divided into 3 groups. Group I received only saline and was evaluated as the control group, whereas Group II received 1/50 percent of LD50 dose of methyl parathion and Group III received 1/20 percent of LD50 dose of methyl parathion every day at 130 pm orally by gavages during two estrus cycles (8 days). The rats at proestrus stage on the morning of 9th day of the study underwent bilateral ovariectomy. Ovarian tissues of the control and drug groups were examined under the electron microscope; primordial and growing follicles were included in the evaluation, however, corpora lutea were excluded taking into account the presence of remaining regressive corpora lutea from the previous cycles.RESULTS:Following examination of ovarian tissues of rats exposed to 1/50 and 1/20 percent of LD50 dose of Methyl parathion at electron microscopic level, it was detected that significant structural changes had occurred in developing follicles and ovarian stroma in both drug groups, and that primordial follicles had not been affected significantly from methyl parathion but necrosis had been developed in oocyte and granulosa cells of developing follicles, and that in 1/20 group in addition to these changes, apoptotic changes had been found in granulosa cells of developing follicles.CONCLUSIONS:As a result of chronic exposure to methyl parathion, rat ovaries are significantly affected and follicular development is impaired. This state may explain the cause of infertility due to chronic pesticide exposure.

Objective: To investigate the clinical significance of mesh peritonization in an experimental rabbit model.Study design: Twenty rabbits were randomly assigned to two groups. A sacrouteropexy operation was performed in both groups using a polypropylene mesh. In the first group, the pelvic peritoneum was not closed over the mesh, and in the second group, the mesh was buried throughout the pelvic retroperitoneal tunnel. One month after mesh implantation, the abdomen was opened and the previous surgical site was explored. The primary outcome was intraabdominal adhesion formation; the secondary outcome was the histologic degree of tissue remodeling.Results: There were no statistically significant differences between the two groups in terms of adhesion scores and collagen organization (P = 0.692, P = 0.097, respectively). There was a greater degree of inflammation in the second group as identified by significantly higher scores for eosinophils-neutrophils, macrophages-foreign body giant cells and mononuclear cells (P = 0.002, P = 0.001 and P = 0.002, respectively).Conclusions: Similar adhesion and collagen organization patterns were found in both groups, but indicators of the inflammatory process were significantly higher in the second group. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

OBJECTIVE:The aim of this study was to investigate the effect of edaravone on experimentally induced ovarian torsion/detorsion ischemia/reperfusion (I/R) injury.STUDY DESIGN:Forty-six female adult Wistar-Albino rats were utilized to create five groups: In group 1, only 5 mg/kg edaravone was given and ovary torsion was not performed. In group 2, torsion was not performed and no drug was given. In group 3, vehicle was given and torsion/detorsion was performed. In group 4, 1 mg/kg edaravone was given and torsion/detorsion was performed. In group 5, edaravone; 5 mg/kg drug was administered and torsion/detorsion was performed. Right ovarian torsion was simulated for a 3-h period of ischemia and a 1-h reperfusion period. Right ovaries were then surgically extirpated in all groups. In ovarian tissue samples malondialdehyde (MDA) levels and activity of superoxide dismutase were studied. Microscopic ovarian tissue damage was scored by histologic and electron microscopic findings.RESULTS:The MDA level in the group 5 was significantly lower than group 3 (p<0.001). Superoxide dismutase activity in the group 5 was significantly higher than group 3 (p<0.001). Histopathological ovarian tissue damage in the group 5 were significantly lower than group 3 (p<0.001).CONCLUSION:These results indicate that edaravone could be an effective agent in the short-term treatment and prevention of ovarian ischemia and reperfusion damage.

Normal female fertility depends on normally occuring oogenesis and maturation progress. Oogenesis and folliculogenesis are different progresses but occure in a harmony and at the same time. Oogenesis includes the events that take place matur ovum produced from primordial germ cells. Although folliculogenesis includes the stages primordial, primary, secondary, matur (Graaf) follicules in the influece of gonadotropines and local growth factors. During oocyte maturation meiosis is distrupted till the puberty. Under LH influence it starts again and first meiosis completes before ovulation. Oocyte maturation can be regarded as the process of coming metaphase II from prophase I of oocyte at the puberty and can be studied as nuclear and cytoplasmic maturation. Meiosis is completed when fertilization occures and zygot is formed. In this article oogenesis, folliculogenesis and oocyte maturation process are summerized with related studies and reiews are revised. [Archives Medical Review Journal 2009; 18(4.000): 227-240]

Normal female fertility depends on normally occuring oogenesis and maturation progress. Oogenesis and folliculogenesis are different progresses but occure in a harmony and at the same time. Oogenesis includes the events that take place matur ovum produced from primordial germ cells. Although folliculogenesis includes the stages primordial, primary, secondary, matur (Graaf) follicules in the influece of gonadotropines and local growth factors. During oocyte maturation meiosis is distrupted till the puberty. Under LH influence it starts again and first meiosis completes before ovulation. Oocyte maturation can be regarded as the process of coming metaphase II from prophase I of oocyte at the puberty and can be studied as nuclear and cytoplasmic maturation. Meiosis is completed when fertilization occures and zygot is formed. In this article oogenesis, folliculogenesis and oocyte maturation process are summerized with related studies and reiews are revised.

Background/Aims: To evaluate by electron microscopy the effects of methotrexate on the tubal morphology of rabbits. Methods: Biopsies were taken from the isthmic portion of both tubes of 4 rabbits in the control group. 1 mg/kg methotrexate was injected in the left tubes and as a control, a 14-gauge fine needle was used to puncture the right tubes of 10 rabbits in the study group. One month later, tubal biopsies were taken from both tubes of every rabbit in the study group. To mimic pregnancy, 100 IU hCG was injected intramuscularly to every rabbit 24 h before every surgery. On examination by electron microscopy, the effects of methotrexate and of the damage with the fine needle on the tubes were compared to the control group. Results: Young epithelial cells of the methotrexate-injected fallopian tubes kept their normal structure, but others contained important ultrastructural changes. These were: nuclear pyknosis, cytoplasmic vacuolization, dilatation of endoplasmic reticulum cisternae, increased cytoplasmic density and compound cilia. Conclusion: Degenerative changes on epithelial cells caused by the temporary blockage of mitotic activity associated with local methotrexate injection are reversible through the formation of new epithelial cells.

Kahraman, S. 212 Kamei, T. 24 Kanayama, S. 133 Karger, T. IV Karlikaya, G. 212 Karša, M. 222 Kavanagh, J.J. 35 Kawaguchi, R. 133 Kaya, M. 217 Kikuchi, A. 73, 116 Kim, D.Y. 62 Kim, H.S. 62 Kim, J.C. 62 Kim, J.W. 41 Kim, S.K. 41 Kim, S.W. 123 Kim, Y.-H. 62 Kitanaka, T. 133 Kliucinskas, M. 32 Kobayashi, H. 133 Kollender, Y. 89 Kostirova, M. 47 Kozuma, S. 116 Kriplani, A. 84 Ku, J.H. 62, 123 Kulaš, T. 222 Kumbak, B. 212

We investigated the ultrastructural effects of methamidophos and the positive effects of 2-pralidoxime (2-PAM) on the liver. Male Wistar-albino rats were assigned to 4 groups and all were treated per os: Group 1 (n=10) received 30 mg/kg methamidophos; Group 2 (n=7) (serving as controls for Group 1) received physiologic NaCl; Group 3 (n=10) received 30 mg/kg methamidophos and was treated with 2-PAM and atropine when cholinergic symptoms were noted; and Group 4 (n=7) (serving as controls for Group 3) was treated with physiologic NaCl. Plasma cholinesterase was measured using radioimmunoassay. Liver tissues were prepared for electron microscopic studies. Methamidophos treatment of Group 1 led to serious changes in hepatocytes and organelles. These changes were not detected in Group 3. In Group 1, the chromatin content of some hepatocyte nuclei and cytoplasmic density increased; these cells also became vacuolar in appearance as a result of lysis in the mitochondrial matrices. In some cells, the lipid content constituted the majority of the cytoplasm. Furthermore, these cells were surrounded by glycogen accumulation. In some areas of the perisinusoidal zone, collagen fibers had increased to form bands. None of these changes were noted in Group 3. These findings suggest that acute organophosphate poisoning causes serious histopathological effects in rat liver, but that these changes are reversible with appropriate treatment strategies.

PURPOSE:To investigate the acute ultrastructural changes that may occur in the contralateral nonischemic lung tissue after unilateral ischemia of a lung in a rabbit model.METHODS:The animals were divided into three main groups of eight; namely, a 2-h procedure group, a 4-h procedure group, and an 8-h procedure group. Each of these groups was further divided into two subgroups of four rabbits each; namely, a control group, given a sham operation without any ischemic insult, and an ischemia group, in which the main pulmonary arteries, the pulmonary veins, and the main bronchi of the left lungs were ligated after thoracotomy. Tissue samples were taken from the left and right lungs to examine the ultrastructural changes after 2, 4, and 8 h of ischemia. Each sample was given a semiquantitative histological injury score. Statistical analysis was done by the Mann-Whitney U-test.RESULTS:Contralateral ultrastructural damage, evident by heterochromatin in the nuclei, mitochondrial degeneration, cisternal widening of the endoplasmic reticulum, increased lipid droplets, and lysosomes, was determined by electron microscopy after unilateral lung ischemia. The contralateral lung injury was significantly correlated with the duration of ischemia.CONCLUSIONS:Unilateral lung ischemia affected the bilateral lungs in a rabbit model. Therefore, in operations such as single-lung transplantation, pulmonectomy, or lobectomy, if the procedure is unnecessarily prolonged, the contralateral lung may be damaged, which could seriously affect the prognosis of the patient.

This study was designed to investigate the preventive effect of melatonin on doxorubicin's most important side effect, cardiotoxicity. Forty male albino Wistar rats were utilized and the rats were divided into five groups: group I, 0.9% NaCl for 4 days; group II, doxorubicin 3 mg/kg/day for 4 days; group III, 2.5 % ethanol for 15 days; group IV, melatonin 6 mg/kg/day for 15 days; and group V, a doxorubicin and melatonin combination were administered intraperitoneally. At the end of the experiment, tissue samples obtained from the cardiac muscle of the left ventricle of the rats were processed for measurement of malondialdehyde and for electron microscopic examination. Malondialdehyde, a product of lipid peroxidation, was found to be significantly higher in the doxorubicin group. However, in the doxorubicin and melatonin combination group the level of malondialdehyde was decreased statistical significant. The histological examination revealed destruction of myofibrils, disorganization of sarcomeres, mitochondrial degeneration and formation of giant mitochondria and lipid accumulation in the doxorubicin group. Also, accumulation of filamentous structures in the sarcoplasma in some of the cells, structural changes in capillaries and an increase in collagen fibers forming bundles were observed. When melatonin was added to the doxorubicin treatment all structural changes were reduced. The cardiotoxic side effect of doxorubicin used as a chemotherapeutic agent and was probably developed as a result of suppression of the antioxidant system and lipid peroxidation. Therefore, it could be assumed that the addition of melatonin in the treatment of doxorubicin could prevent the cardiotoxicity of doxorubicin.

The antigen presenting cells (APCs) with special interest to dendritic cells (DC), were investigated in 28 hypertrophic and 10 control pharyngeal tonsils of children by histochemistry, immunohistochemistry and electron microscopy. In this study, we are trying to clarify the function and classification of APC in pharyngeal tonsils using morphologic criteria, Human Leukocyte Antigen Monoclonal Antibody (HLA-DR MoAb), which is specific for APCs, and acid phosphatase (APh) reacting with both phagosomes and lysosomes. The surface epithelium of the patient group examined by light microscopy, heavy infiltration of lymphocytes, degenerated columnar cells and a few HLA-DR MoAb (+) columnar cells was observed. Additionally, a significant number of APCs which were Langerhans cells (LCs), interdigitating dendritic cell (IDC), follicular dendritic cell (FDC) and macrophages were stained with both HLA-DR MoAb and APh in the epithelial, interfollicular-subepithelial and follicular areas. Ultrastructural examinations revealed that lymphocytes, macrophages, LC and M cells were found among the surface columnar epithelial cells of the patient group. The interactions between M cells and LC suggested that M cells probably passed antigens from surface to LC. In the interfollicular-subepithelial areas of the hypertrophic pharyngeal tonsil, IDCs were in close contact with lymphocytes, macrophages and plasma cells. Seven types of FDCs (FDC-1 - FDC-7) were recognised according to their ultrastructural appearances. Differentiated FDCs (FDC-4) were also in close contact with each active subtype of FDCs in follicular areas besides lymphocytes. These findings supported the idea that although the pharyngeal tonsils contained several types of active APCs, only DC were in close contact with immunocompetent cells and the other APC's. Therefore, these morphologic appearances of DC could be a sign of function to initiate the immune response of the pharyngeal tonsil.

While it is well known that unilateral tissue ischemia may result in contralateral damage in some paired organs, there is no universally accepted mechanism to explain why these contralateral changes occur. The aim of the present study was to investigate the ultrastructural and hormonal changes that occur in the contralateral nonischemic adrenal gland after unilateral ischemia of an adrenal gland in a rat model. The animals were divided into four groups of four rats each; namely, a control group which received a sham operation without any ischemic insult, a 2-h ischemic group, a 4-h ischemic group, and an 8-h ischemic group. The left adrenal blood vessels were ligated in all ischemia groups and blood samples were taken for hormonal study 2, 4, and 8 h later, after which bilateral adrenalectomy was performed to determine the ultrastructural changes. The plasma concentrations of aldosterone and cortisol were determined by radioimmunoassays. There was an increase in both aldosterone and cortisol levels related to the duration of the ischemia, but the differences among the groups were not statistically significant. Contralateral ultrastructural damage such as heterochromatin in nuclei, mitochondrial degeneration, endoplasmic reticulum cisternal widening, increased lipid droplets, and lysosomes, were demonstrated electron-microscopically after unilateral adrenal ischemia.

PURPOSE:Adenoidectomy, especially for the treatment of suppurative otitis media, has been used for a very long time. In this study, the role of adenoids in the origin of otitis media with effusion was investigated by using light microscopy, immunocytochemistry, enzyme chemistry, and electron microscopy. MATERIALS AND METHODS:A group of 28 children with otitis media with effusion (OME) was identified. Ages ranged from 3 to 12 years. A control group of 10 age-matched children without any middle ear and upper respiratory tract infection served as the basis for comparison. Specimens obtained at surgeries from both groups were divided into groups for light microscopy, immunocytochemistry, enzyme cytochemistry, and electron microscopy and then all were examined blindly. Also, quantitative analysis of antigen-presenting cells was performed blindly on 10 patients and 10 controls. RESULTS:There was an increase in the number of lymphocytes, mast cells, plasma cells, macrophages, dendritic cells, and M cells in the adenoids of patients with OME when compared with the normal cases. Stratified squamous epithelial areas, collagenous fibers, and fibrocytes were also increased in the patient group. Antigen-presenting functions of epithelial cells are shown by major histocompatibility complex (MHC) class II positivity of some ciliated-columnar epithelial cells in the patient group. CONCLUSION:Adenoid tissues of patients with OME in this study seem to be infectious foci, aggravating immune reactions, which might attack the middle ear through an ascending route.