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Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore
2021
Mariângela Bueno Cordeiro Maldonado | Francine Messias Ciríaco Henry | Teissiane Fernanda de Vasconcelos Ferreira | Barbara Piffero Mello | Mario Binelli | Claudia Maria Bertan Membrive
The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 μM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 μM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17β-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.
Mostrar más [+] Menos [-]Toxicological evaluation of flumequine in pubertal male rats after oral administration for six weeks
2018
Kang, JeongWoo | Hossain, Md Akil | Choi, Byungkook | Cho, Joon-Hyoung | Kang, Seok-Jin | Ku, Hyun-Ok | Jeong, Sang-Hee | Kang, Hwan-Goo
Veterinarians use flumequine (FLU) widely but its toxicological effects are still unclear. FLU doses of 53, 200, or 750 mg/kg were administered orally for six weeks to pubertal male rats for evaluation of their toxicity. Weight gain was poorer after seven days of exposure to FLU 750, but relative weights of the brain, adrenal and thyroid glands, and testes were notably higher. Haematological and lipid profile parameters, cardiac markers, and inorganic phosphate significantly increased in the FLU 750 group. Blood glucose, oestradiol and serum concentrations of immunoglobulins G (IgG) and E (IgE) significantly decreased after treatment. The levels of interleukins 10 (IL-10) and 6 (IL-6) fell significantly in the FLU 200 and FLU 750 groups. Cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) and cyclooxygenase-2 (Cox-2) expression amplified after treatment. Serum levels of free triiodothyronine (fT3) and free thyroxine (fT4) reduced in the FLU 200 and FLU 750 groups without changes in total T3 or T4 level. All doses of FLU significantly depressed concentrations of thyroid-stimulating hormone (TSH) and testosterone. Histopathology of thyroid glands from rats treated with FLU 750 showed degeneration and depletion of thyroid follicular epithelial cells. Expression of 8-hydroxydeoxyguanosine (8-OHdG) was increased in a dose-dependent manner in the brain, but decreased in the testes. Expression of CYP1A1 increased in the adrenal and pituitary glands. The results of this study suggest that the toxicity of FLU in rats is an effect of its disruptive influence on the pituitary-thyroid hormonal system and on the dysfunction of the immune system.
Mostrar más [+] Menos [-]Monitoring vaginal electrical resistance, follicular waves, and hormonal profile during oestrous cycle in the transition period in Bangladeshi sheep
2018
Talukder, Mohammad Rafiqul Islam | Hasan, Moinul | Rosy, Tasmia Akter | Bari, Farida Yeasmin | Juyena, Nasrin Sultana
Introduction: The ovarian follicular dynamics, vaginal electrical resistance (VER), progesterone (P4) and oestrogen (E2) profiles were investigated during the oestrous cycle in four indigenous ewes. Material and Methods: Daily VER values were recorded with a heat detector. The follicles were observed and measured by trans-rectal ultrasonography. Blood was collected daily for hormonal profiles. Results: A significant variation in VER values (P < 0.05) in oestrus by ewes and position in the sequence of cycles was observed. Trans-rectal ultrasonography of ovaries revealed the presence of 2–4 waves of follicular growth. Study of hormonal profiles by ELISA revealed a positive correlation between E2 concentration and development of follicles and a negative correlation between P4 concentration and their development. The concentrations of oestradiol increased in oestrus and then decreased to a basal level. Follicular growth was accompanied by a rise in the concentration of serum oestradiol. Inversely, when follicles received the stimulation for ovulation, concentration of progesterone started to fall, but after ovulation, it climbed back to its peak and remained at this state until next ovulatory follicle reached its maximum diameter. Conclusion: This study could help to set up a manipulative reproductive technique for improving genetic values in indigenous sheep.
Mostrar más [+] Menos [-]Levels of the natural hormones 17β-oestradiol and testosterone in serum of cattle: results from population studies in Poland
2016
Woźniak-Sobczak, Barbara | Witek, Sebastian | Matraszek-Żuchowska, Iwona | Kłopot, Alicja | Posyniak, Andrzej
Introduction: The present study is a comprehensive overview of the natural occurrence of 17β-oestradiol and testosterone in serum of cattle in Poland. Material and Methods: The serum samples (n = 826) were collected from cattle within five years. The samples were examined for the presence of oestradiol and testosterone using ELISA or gas chromatography with mass spectrometry. Results: In 98 samples (24%) 17β-oestradiol was detected above decision limits of applied methods, including five samples over the recommended concentration of 0.1 μg L⁻¹. Of the serum samples taken from cows (≤18 months of age), 95 and 99 percentiles of the animals had 17β-oestradiol concentration below 0.027 and 0.086 μg L⁻¹ and of samples from cows over 18 months of age - below 0.059 and 0.125 μg L⁻¹ respectively. Calculated values for bulls (≤18 months of age) were 0.025 and 0.034 μg L⁻¹ and for the animals older than 18 months of age - 0.035 and 0.041 μg L⁻¹. The natural presence of testosterone was detected in 201 serum samples (48.7%). According to the obtained data, 95% and 99% of cows (≤18 months of age) serum samples had testosterone concentration below 0.05 and 0.23 μg L⁻¹ and the animals over 18 months of age - 0.30 and 0.49 μg L⁻¹, respectively. For bulls these values did not depend on the age of the animals and were in the ranges of 5 - 6.3 μg L⁻¹ (95%) and 11.4 - 12.1 μg L⁻¹ (99%). Conclusion: Our study showed that the threshold values for these hormones in plasma of cattle designated years ago are correct, but they need to be supplemented for animals older than 18 months.
Mostrar más [+] Menos [-]Effects of α-enolase gene silencing on reproductive-related hormone receptor expression and steroid hormone synthesis of primary granulosa cells from goose F1 follicles
2020
Ji, Hong | Niu, Chun-Yang | Zhang, Hong-Liang | Guo, Jing-Ru | Zhen, Li | Lian, Shuai | Yang, Chuang | Yang, Huan-Min | Wang, Jian-Fa
Enolases are enzymes in the glycolytic pathway, which catalyse the reversible conversion of D-2-phosphoglycerate into phosphoenol pyruvate in the second half of the pathway. In this research, the effects of α-enolase (ENO1) on steroid reproductive-related hormone receptor expression and on hormone synthesis of primary granulosa cells from goose F1 follicles were studied. Primary granulosa cells from the F1 follicles of eight healthy 8-month-old Zi geese were separated and cultured. An ENO1 interference expression vector was designed, constructed and transfected into primary cultured granulosa cells. The mRNA expression levels of follicle-stimulating hormone receptor (FSHR), luteinising hormone receptor (LHR), oestrogen receptor α (ER α), oestrogen receptor β (ER β), growth hormone receptor (GHR) and insulin-like growth factor binding protein-1 (IGFBP-1) in the cells were evaluated as were the secretion levels of oestradiol, activin, progesterone, testosterone, inhibin and follistatin in cell supernatant. α-enolase gene silencing reduced the expression of FSHR, LHR, ERα, ERβ, GHR, and IGFBP-1 mRNA, potentiated the secretion of oestrogen, progesterone, testosterone, and follistatin of granulosa cells, and hampered the production of activin and inhibin. ENO1 can regulate the reactivity of granulosa cells to reproductive hormones and regulate cell growth and development by adjusting their hormone secretion and reproductive hormone receptor expression. The study provided a better understanding of the functional action of ENO1 in the processes of goose ovary development and egg laying.
Mostrar más [+] Menos [-]Effect of estradiol benzoate injection to male rabbits on glucose, total protein, albumin, calcium concentrations and prostate tissue
2017
Tamour Elkhier | Shams-Eldein Hassaballa | Shadia Abdo-Elatti Omer | Abdelnasir Adam
Objective: This study was carried out to investigate the effects of estradiol benzoate (EB) injection to male rabbits on glucose, total-protein, albumin, calcium concentrations and prostate tissue, and the role of prolactin as an important mediator of estrogen action in prostate. Materials and methods: Fifty four adult male rabbits were used in this study. The rabbits were randomly divided into two groups. Group A contained 36 male rabbits, which were further randomly divided into four sub-groups, three of them contained 10 rabbits and one sub-group contained 6 rabbits as control. Group B contained 18 male rabbits, which were divided randomly into three equal sub-groups. Three sub-groups of Groups A and B were treated once each on alternative day with the intramuscular injections of EB dosed at 40, 80 and 120 µg/rabbit, respectively for 20 days, whereas the fourth sub-group of Group A received no estradiol, and Group B received 1 mg Bromocriptine Mesilate in addition to EB through oral route on each alternative day. Blood samples were collected for measuring glucose, t-protein, albumin and calcium levels. Prostate tissue samples were collected from all the rabbits for histological studies. Results: Glucose was significantly (P˂0.05) increased as a result of 80 µg EB injection, while significantly (P˂0.05) decreased due to 40 and 120 µg EB injection. Total protein significantly (P˂0.05) increased due to injection of 40 µg EB, however t-protein was not changed due to 40 and 120 µg injection. On the other hand, the results of albumin and calcium were not affected (P˃0.05) by EB. In prostate tissues, EB induced hyperplasia with dysplasia or dysplasia only, but this effect was mild due to inhibition of prolactin.Conclusion: The injection of EB to male rabbits increased or decreased glucose level, increased t-protein level mildly or not changed, while albumin and calcium levels were not affected. EB induced hyperplasia on prostate tissue, and this effect was reduced by prolactin inhibition indicating that prolactin might have a role on the action of estrogen. [J Adv Vet Anim Res 2017; 4(1.000): 15-21]
Mostrar más [+] Menos [-]Ultrasonographic and hormonal characterization of reproductive health and disease in wild, semiwild, and aquarium-housed southern stingrays (Hypanus americanus)
2019
Mylniczenko, Natalie D. | Sumigama, Shiho | Wyffels, Jennifer T. | Wheaton, Catharine J. | Guttridge, Tristan L. | DiRocco, Stacy | Penfold, Linda M.
OBJECTIVE To characterize physical examination, plasma biochemical, and ultrasonographic findings in aquarium-housed, managed semiwild, and wild southern stingrays (Hypanus americanus) with and without reproductive disease. ANIMALS Southern stingrays from aquarium (n = 48), lagoon (managed semiwild; 34), and wild (12) habitats. PROCEDURES Limited, opportunistic prosections were performed of presumed anatomically normal wild southern stingrays and compared with findings for aquarium-housed stingrays with reproductive disease. Ultrasonographic video data from both groups were used to assign a score (1 to 5) indicating increasing severity of ovarian and uterine reproductive disease. Plasma total 17β-estradiol, estrone, progesterone, and testosterone concentrations were measured with enzyme immunoassays validated for use in southern stingrays. RESULTS Ultrasonographic ovarian scores were significantly correlated with uterine scores. No reproductive disease was detected in semiwild or wild stingrays, but 65% (31/48) of aquarium-housed stingrays had developing or advanced reproductive disease (ie, ultrasonographic ovarian or uterine score of 4 or 5). Significant correlations were identified between ovarian and uterine disease status and plasma concentrations of all steroid hormones except testosterone. CONCLUSIONS AND CLINICAL RELEVANCE Findings suggested that ultrasonography and plasma hormone concentrations may be useful in the identification of reproductive disease and determination of disease severity in southern stingrays.
Mostrar más [+] Menos [-]Changes in luteinizing hormone secretion after estradiol treatment in prepubertal Nelore heifers
2011
Cardoso, Daniel | Guerra, Fábio F. | Peiró, Juliana R. | Perri, Silvia H.V. | Nogueira, Guilherme P.
Changes in luteinizing hormone (LH) secretion after 17beta-estradiol (E2) injection were evaluated during sexual maturation in 10 prepubertal Nelore heifers. Heifers were divided into 2 groups: intact (I) and ovariectomized (OVX). 17beta-estradiol (2 micrograms/kg) was administered to both groups at 10, 13, and 17 mo of age. Only at 10 mo of age was there a greater mean LH concentration in OVX heifers (1.33 +/- 0.29 ng/mL) compared with the I group (0.57 +/- 0.15 ng/mL). At 13 and 17 mo of age there was no significant difference between the 2 groups in any of the evaluated variables (number of peaks, total peak area, greatest peak area, and time to greatest peak occurrence). This suggests a decrease in negative E2 feedback associated with an increase in positive feedback to LH secretion during sexual maturation, and these were likely the key factors that determined the time of first ovulation in Nelore heifers.
Mostrar más [+] Menos [-]Efficacy of domperidone and sulpiride as treatments for fescue toxicosis in horses
1994
Redmond, L.M. | Cross, D.L. | Strickland, J.R. | Kennedy, S.W.
We evaluated the effectiveness of 2 dopamine antagonists as treatments for fescue toxicosis in horses. Sixteen gravid mares were assigned by breed and expected foaling date to 1 of 3 treatment groups: endophyte-infested control 1.1 mg of domperidone/kg of body weight/d; and 3.3 mg of sulpiride/kg/d. Mares were pastured on endophyte-infected fescue and received 0.454 kg of a corn and dried molasses carrier containing the drug treatment. Treatment started 30 days prior to expected foaling date and continued until parturition. Blood samples were collected, and mammary gland scores were recorded every 5 days. Body weight and body condition scores were obtained every 28 days. Serum was analyzed for prolactin, progesterone, and estradiol-17beta concentrations. Domperidone-treated mares had shorter (P = 0.09) gestation duration and foaled closer (P = 0.07) to their expected parturition date than did control mares. Mammary gland scores were higher (P < 0.05) for domperidone-treated mares than for control mares. By 4 and 9 days after the start of treatment, serum prolactin concentration was higher P < 0.05) in domperidone-treated mares and sulpiride-treated mares, respectively, than in control mares. Domperidone- and and sulpiride-treated mares had higher (P < 0.05) serum progesterone and lower (P < 0.01) estradiol-17beta concentrations than did control mares. These results indicate that domperidone may offer considerable potential as a treatment for fescue toxicosis in horses.
Mostrar más [+] Menos [-]Effect of mimicking prepartum concentration of estradiol-17 beta on the inflammatory response to endotoxin in gilts
1994
Magnusson, U. | Holst, H. | Kindahl, H. | Karlsson, A.
The effect of mimicking prepartum concentration of estradiol-17 beta on the inflammatory response to endotoxin in gilts was studied. The study was performed in a split-litter design and comprised 5 pairs of littermates. A catheter was inserted into the jugular vein 2 days prior to the start of the study. In each pair, 1 littermate was treated IM with 2.5 mg of estradiol-17 beta/75 kg of body weight, and the other littermate was given peanut oil IM as a control. The day after treatment, all gilts were challenge-exposed with a Salmonella typhimurium-derived endotoxin (1 microgram/kg, IV) and the inflammatory response to challenge exposure was monitored. There was no effect of estradiol treatment on the transient clinical signs of endotoxemia or on the increase in rectal temperature. The increase in blood concentrations of prostaglandin F2 alpha, metabolite and cortisol after endotoxin challenge exposure was not affected by estradiol. Decrease in number of circulating blood mononuclear cells and polymorphonuclear leukocytes was not changed by estradiol treatment. Taken together, mimicking prepartum concentration of estradiol did not affect either the magnitude or the kinetics of the inflammatory response to endotoxin in gilts. Relevance of these findings to development of endotoxin-mediated diseases, such as the postpartum agalactia syndrome, needs further study.
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