The protective effect of vitamin E on rats’ ovarian follicles following an administration of diazinon: An experimental study

Background: Diazinon (DZN) is an organophosphate insecticide that has been widely utilized in agriculture all over the world and caused many negative effects on different species such as plants and animal species, especially on a human. Objective: The aim of the present study was to evaluate the protective effect of vitamin E on rats’ ovarian follicles following an administration of diazinon. Materials and Methods: A total of 30 adult female Wistar rats were divided into five groups: a control group (without any intervention), sham group (received only pure olive oil, as solvent), experimental group I (DZN+olive oil, 60 mg/kg), experimental group II (vitamin E, 200 mg/kg), and experimental group III (DZN: 60 mg/kg+vitamin E: 200 mg/kg). All drugs were injected intraperitoneally, except vitamin E which was administrated by gavage. The animals were sacrificed after two weeks and the left ovary was used to measure proliferation of ovarian follicles. Tissues were analyzed by the PCNA technique and viewed with an optical microscope for evaluating cell proliferation. Results: The result of the present study revealed that the number of proliferative cells in the experimental group I decreased significantly in contrast to the control group in secondary and Graffian follicles (p< 0.001). The administration of vitamin E plus DZN significantly increased proliferative cells compared to the DZN group (p< 0.001). Primordial follicles showed that all study groups were lacking PCNA positive cells, which means no expression of PCNA in these follicles. The results of this study showed that primary follicles in all study groups had a few and scattered PCNA positive cells with no significant difference between the groups (p> 0.05). Conclusion: Results showed that DZN reduced proliferation in secondary and Graffian follicles and vitamin E increased it. The results of this study suggested that vitamin E by its antioxidant activity was able to improve the DZN-induced ovarian toxicity. Key words: Diazinon, Proliferation, Ovary, Vitamin E, Rat

Effects of monosodium glutamate on apoptosis of germ cells in testicular tissue of adult rat: An experimental study

Background: Monosodium glutamate (MSG) is used as a flavoring and food seasoning. Some studies have reported the oxidative effects of using this substance on various tissues. Objective: This study has investigated the effects of MSG and the protective effect of vitamin C (vit C) on apoptosis of testicular germ cells and biochemical factors. Materials and Methods: In this experimental study, 24 adult male Wistar rats were randomly divided into four groups: control (received distilled water), vit C group (150 mg/kg), experimental group 1 (MSG 3 gr/kg), experimental group 2 (MSG 3 gr/kg + vit C 150 mg/kg). The rats were gavaged for 30 days, and then were sacrificed, the right testis was isolated for biochemical examinations for the glutathione, malondialdehyde, and left testis used in histological experiments. Tunnel staining was used to determine the number of apoptotic cells. Results: The results showed that apoptotic cells in the MSG group had a significant increase compared to the control group (P = 0.001), but the number of these cells in the MSG co-administered with vit C and vit C groups were significantly lower than the MSG group. Germinal epithelial thickness also decreased in MSG group compared to the control group. Conclusion: MSG can lead to increase apoptotic changes in the germinal epithelial of the testicle, and vit C as an antioxidant can modify the pathological and biochemical changes induced by MSG

Effects of Hypothyroidism on Proliferation and Programmed Cell-Death in Rat Ovarian Granulosa Cells

Hypothyroidism is the most common endocrine disease, after diabetes. Thyroid hormones are essential for genital organs function. In this study, we aimed to determine the apoptotic and cell proliferation indexes resulting from reduced thyroid hormones in rat ovarian follicles. For the purpose of this study, 20 female mature Wistar rats were divided into test and control groups. The test group underwent chemical thyroidectomy by receiving 500 mg/l propylthiouracil added to drinking water. The control group only received ordinary drinking water. After three weeks, the rats were sacrificed and their ovaries were removed and fixed for tissue preparation. Triphosphate-biotin nick end-labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) immunohistochemical techniques were applied to determine apoptosis and cell proliferation variations. Our findings revealed that apoptotic index significantly diminished in large antral follicles. There were no significant differences between the two groups in terms of primary and pre-antral follicles. No TUNEL-positive cell was noted in primordial follicles in the both groups. Cell proliferation index revealed a significant decrease in follicular growth of pre-antral to graafian follicles in the hypothyroid group. PCNA-positive cells were not observed in primordial follicles in the both groups. The results of the study suggested that reduced thyroid hormones lead to a wide range of hormonal changes, and factors existing in follicular fluid, especially in large antral follicles, undergo transformations that affect apoptotic and cell proliferation indexes. The process of follicular growth occurs by entrance of follicles to the next growth phase without inducing sufficient potentiality, and the produced ovules might be healthy or morphologically defected

Effects of Hypothyroidism on Proliferation and Programmed Cell-Death in Rat Ovarian Granulosa Cells

Hypothyroidism is the most common endocrine disease, after diabetes. Thyroid hormones are essential for genital organs function. In this study, we aimed to determine the apoptotic and cell proliferation indexes resulting from reduced thyroid hormones in rat ovarian follicles. For the purpose of this study, 20 female mature Wistar rats were divided into test and control groups. The test group underwent chemical thyroidectomy by receiving 500 mg/l propylthiouracil added to drinking water. The control group only received ordinary drinking water. After three weeks, the rats were sacrificed and their ovaries were removed and fixed for tissue preparation. Triphosphate-biotin nick end-labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) immunohistochemical techniques were applied to determine apoptosis and cell proliferation variations. Our findings revealed that apoptotic index significantly diminished in large antral follicles. There were no significant differences between the two groups in terms of primary and pre-antral follicles. No TUNEL-positive cell was noted in primordial follicles in the both groups. Cell proliferation index revealed a significant decrease in follicular growth of pre-antral to graafian follicles in the hypothyroid group. PCNA-positive cells were not observed in primordial follicles in the both groups. The results of the study suggested that reduced thyroid hormones lead to a wide range of hormonal changes, and factors existing in follicular fluid, especially in large antral follicles, undergo transformations that affect apoptotic and cell proliferation indexes. The process of follicular growth occurs by entrance of follicles to the next growth phase without inducing sufficient potentiality, and the produced ovules might be healthy or morphologically defected

Effects of Bene (Pistacia atlantica) on Histopathology of Testis, Sperm Chromatin Quality and Stress Oxidative in Busulfan-Induced Infertile Mice

Background: Some plants stimulate spermatogenesis and increase fertility, while some cause spermatogenesis arrest. So far, the effects of bene (Pistacia atlantica) on male fertility have not been studied. The aim of this study was to investigate the effects of bene on sperm parameters, testicular histopathology, sperm quality, and oxidative stress in busulfan-induced infertile mice. Methods: Thirty-five male BALB/c mice were randomly assigned to control, sham, busulfan, bene, and bene + busulfan groups. The busulfan group received 10 mg/kg as a single dose and intraperitoneally. The bene group received pellets containing 10% of bene. Another group received 10 mg/kg busulfan and was fed with pellet containing 10% bene. Then, sperms, sperm chromatin quality, testicular histopathology, and oxidative stress levels were studied on the 35th day of the experiment. Results: Busulfan injection resulted in a significant reduction in sperm parameters compared to the control group (p<0.001); it decreased after bene administration (p<0.001). In addition, in the group treated with bene, the sperm count with damaged DNA was reduced and the level of malondialdehyde decreased compared to the busulfan group. A significant increase was observed in the mean level of superoxide dismutase and catalase enzymes in the bene + busulfan group compared to the busulfan group (p<0.001). The histopathological improvement of the testis was observed in the bene + busulfan group. Conclusion: The administration of 10 mg/kg of bene powder for 35 days reduced the oxidative stress, improved testicular histopathology, sperm chromatin quality, and sperm parameters in the infertile mice model

Assessment of sperm morphology, chromatin integrity, and catsper genes expression in hypothyroid mice

There is an evident relationship between the fertilizing capacity of sperm and the normal morphology, quality chromatin, and motility of sperm. It is well known that thyroid hormones are the important regulators of testicular function. A correlation was found between the hypothyroidism and sperm damages. The present study was conducted to investigate the effects of hypothyroidism on sperm morphology, chromatin quality, and motility. For this purpose, 20 male mice were divided into the control and the hypothyroid groups that received 0.05% 6-n-propyl-2-thiouracil (PTU) for 35 days. Sperm morphology with Papanicolaou staining and sperm chromatin quality with both Aniline Blue (AB) and Toluidine blue (TB) staining were assessed. Besides, immunohistochemistry and real-time PCR were performed to evaluate the changes of cation sperm channel (CatSper) genes. A significant increase in the sperm chromatin condensation was found in the hypothyroid mice compared to the control mice (p < 0.05). Furthermore, a significant decrease was observed in the morphology of normal sperm in hypothyroid mice compared to the controls (p < 0.05). The results showed that Hypothyroidism could downregulate the expression of CatSper genes. Immunohistochemical data confirmed the real time-PCR results. Furthermore, the results showed that hypothyroidism could adversely affect sperm morphology, sperm chromatin condensation, and CatSper gene expression in mice and these abnormalities may be related to the excessive production of reactive oxygen species (ROS) in a hypothyroid state

P53 expression in various types of hydropic placentas (through ploidy analysis as a complementary tool in diagnosis of samples)

Background: Placentas characterized by hydropic swelling of chorionic villi occur in a spectrum of pathological conditions including hydropic abortion (HA), partial hydatidiform mole (PHM) and complete hydatidiform mole (CHM). The purpose of this study was to investigate whether the expression of p53 tumour suppressor protein could differentiate these various types of hydropic placentas. Methods: p53 immunohistochemical staining was performed in 19 molar (8 PHM and 11 CHM) and 10 non-molar (HA) formalin-fixed, paraffin-embedded tissue samples. Ploidy analysis using flow cytometry was performed as a complementary tool in diagnosis of samples. Results: DNA histograms obtained from all samples had confirmed diploidy in HAs and CHMs and triploidy in PHMs. p53 immunoreactivity was assessed in villous cytotrophoblasts, syncytiotrophoblasts and stromal cells. The p53 positive reaction was predominantly observed in the nuclei of cytotrophoblastic cells and rarely in stromal cells, no reaction was seen in syncytiotrophoblasts. The mean percentage of p53 positive cells were 6.10&#177;3.75 for HA, 25.87&#177;13.4 for PHM and 39.83&#177;18.76 for CHM.&#160; There was a significant difference in P53 immunoreactivity of cytotrophoblastic cells between CHM and HA (P&#60;0.001), and between PHM and HA (P=0.004). There was no significant difference in immunohistochemical reactivity between CHM and PHM (P=0.068). Conclusion: This study confirms that p53 immunostaining may be helpful in distinguishing complete and partial hydatidiform mole from hydropic abortion, but not complete hydatidiform mole from partial hydatidiform mole

Expression of Pro-Apoptotic Bax and Anti-Apoptotic Bcl-2 Proteins in Hydatidiform Moles and Placentas With Hydropic Changes

Morphologic examination still forms the main diagnostic tool in the differential diagnosis of molar placentas. However the criteria are subjective and show considerable inter-observer variability among pathologists. The aim of the present study was to investigate the role of Bcl-2 and Bax immunostaining in the differential diagnosis of molar placentas. Bax and Bcl-2 immunohistochemical staining were performed in 19 molars (8 partial and 11 complete hydatidiform mole) and 10 non-molar (hydropic abortion) formalin-fixed, paraffin-embedded tissue samples. Ploidy analysis using flow cytometry had confirmed diploidy in hydropic abortions and complete hydatidiform moles and triploidy in partial hydatidiform moles. Bcl-2 expression was observed only in syncytiotrophoblasts, No immunoreactivity was detected in Cytotrophoblasts, and stromal cells, the total score averages of Bcl-2 immunoexpression in partial hydatidiform moles and hydropic abortions were significantly higher than in complete hydatidiform moles, whereas no significant difference was observed between partial hydatidiform moles and hydropic abortions. Bax immunoreactivity was observed in cytotrophoblasts, stromal cells and occasionally in syncytiotrophoblasts. No statistically significant difference in Bax immunoexpression total score was observed among various groups. Based on the results of this study, Bcl-2 immunostaining offers a potential adjunctive diagnostic tool to distinguish complete hydatidiform mole from partial hydatidiform mole and hydropic abortion, but not partial hydatidiform mole from hydropic abortion, Bax immunostaining cannot be helpful in this regard

Lectin Histochemical Study of Vasculogenesis During Rat Pituitary Morphogenesis

Objective(s) The aim of this study was to investigate glycoconjugates distribution patterns as well as their changes during the course of pituitary portal vasculogenesis and angiogenesis.Materials and MethodsFormalin fixed paraffin sections of 10 to 20 days of Sprague Dawly rat fetuses were processed for histochemical studies using four different horseradish peroxidase (HRP) conjugated lectins. Orange peel fungus (OFA), Vicica villosa (VVA), Glycine max (SBA) and Wistaria floribunda (WFA) specific for α-L-Fucose, D-Gal, α, ß-D-GalNAc and D- GalNAc terminal sugars of glycoconjugates respectively.ResultsOur finding indicated that adenohypophysal cells reacted with OFA on gestational day 10 (E10) and increased progressively to E14. Staining intensity did not change from days 14 to17, then after increased following days to E20 significantly (P< 0.05). A few cells around Rathke’s pouch reacted with VVA on E13, increased to E14 and decreased significantly afterward (P< 0.05). Reaction of some cells around Rathke’s pouch reacted with SBA on E14. This visible reaction was the same as E18 and decreased later (P< 0.05). Many cells around Rathke’s pouch reacted with WFA on E13 and increased on E 14 and E15 and decreased thereafter (P< 0.05).ConclusionReactions of OFA and other tested lectins with endothelial cells around Rathke’s pouch and developing pars distalis were different. These results suggest that embryonic origin of hypophiseal pituitary portal (HPP) system endothelial cells are not the same and our finding also indicated that glycoconjugates with terminal sugars α-L-Fucose, D-Gal, α, ß-D-GalNAc may play critical role(s) in cell interactions and tissue differentiations such as vasculogensis and angiogenesis as well as other developmental precursors in formation of the pituitary gland

Intonation and Duration Curve in Persian Interrogative Sentences

Background and Aim: prosody is a very important factor in communication and includes such parameters as: duration, intonation, pitch, stress, rhythm etc. Intonation is the pitch variation in one sentence. Duration is the time taken to utter a voice. The aim of the present study was to evaluate some parameters of prosody such as duration and intonation curve in interrogative sentences among normal Farsi speaking adults in order to determine the characteristics of this aspect of language with an emphasis on laboratory testing.Methods: This study was performed as a cross-sectional one. The participants included 134 male and female Farsi speaking individuals aging between 18-30 years. In this study two interrogative sentences with open and closed answers were used. The voice samples were analyzed by Dr.speech -real analysis software. Data analysis incorporated unilateral analysis of variance and an intonation curve was drawn for each sentence.Results: The parameter of duration among men and women was significantly different (p≤0.001). Duration in open questions was significantly longer than yes/no questions (p≤0.001). The intonation curve of the two groups were similar.Conclusion: Men and women use duration changes, for making difference in prosody. On the whole, duration among women is longer than men. In open questions, the duration of sentences is mostly due to the question word. The intonation curve in open questions has more amplitude. Women show much more changes in basic frequency for transferring interrogative state in their expressions