RESUMO
A variety of parameters, including liquefaction and semen viscosity, affect the sperm's ability to travel and reach the egg for fertilization and conception. Given that the details behind the viscosity of the semen in male camels have not yet been fully clarified, the purpose of this study was to ascertain how the addition of papain affected the viscosity of fresh diluted camel semen. The study examined semen samples derived from camels that had distinct viscosities. Sperm motility, viability, abnormal sperm percentage, concentration, viscosity, morphometry, acrosome integrity and liquefaction were among the evaluations following 0, 5, 10, 20 or 30 min of incubation at 37°C with papain (0.004 mg/mL, 0.04 mg/mL or 0.4 mg/mL; a semen sample without papain was used as a control). A statistically significant interaction between the effects of papain concentrations and incubation time was found (F = 41.68, p = .0001). Papain concentrations (p = .0001) and incubation times (p = .0001) both had a statistically significant impact on viscosity, according to a simple main effects analysis. A lower viscosity was found (p < .05) at 0.04 mg/mL (0.1 ± 0.0) after 10 min of incubation. A simple main effects analysis showed that papain concentrations and incubation time have a statistically significant effect on sperm motility (p = .0001). At 0.04 mg/mL papain, the sperm motility % was higher (p < .05) after 10 min (64.4 ± 4.8), 20 min (68.4 ± 6.2), and 30 min incubation (72.2 ± 6.6) compared to 0, 5 min (38.3 ± 4.1 and 51.6 ± 5.0, respectively). In conclusion, the fresh diluted camel semen had the lowest viscosity properties after 10 min of incubation with 0.04 mg/mL papain, without compromising sperm motility, viability, acrosome integrity and sperm morphology.
Assuntos
Camelus , Papaína , Preservação do Sêmen , Sêmen , Motilidade dos Espermatozoides , Animais , Papaína/farmacologia , Masculino , Viscosidade , Motilidade dos Espermatozoides/efeitos dos fármacos , Sêmen/efeitos dos fármacos , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Análise do Sêmen/veterinária , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Acrossomo/efeitos dos fármacosRESUMO
Spermatozoa can experience negative changes when subjected to freezing and thawing, including lowered motility, viability and acrosome response. Herein, the effects of different concentrations of soybean lecithin nanoparticles on cryopreserved Holstein bull semen were examined. Semen was collected, cryopreserved and utilized for sperm kinetic parameter analysis following dilution, equilibration and thawing with 0.5% soybean lecithin (E1), the control extender, and 0.75% (E2), 0.5% (E3), 0.25% (E4) and 0.125% (E5) of lecithin nanoparticles. Results revealed that following dilution, the progressive motility (PM) at E3, E4 and E5 of lecithin nanoparticles was higher (p < .05) than it was for E2. After equilibration, compared to the E1, E2, and E3 values, the PM, vitality, normal morphology, membrane integrity and intact acrosome values at the E5 were consistently greater (p < .05). Comparing the percentages of intact acrosome and membrane integrity at E2 and E3 to E4 and E5, a substantial decrease (p < .05) was seen. Following thawing, the percentage of PM improved at E2 and E5, even though their mean PM values were similar (p > .05) compared to E1, E3 and E4. Vigour and progression parameters of sperm (DAP, DCL, DSL, VAP, VCL, VSL and STR) at E5 were higher (p < .05) than those at E1, E2, E3 and E4. In conclusion, the cryopreserved sperm from Holstein bulls revealed outstanding properties both after equilibration and after thawing with 0.125% lecithin nanoparticles, and they were sensitive to high dosages.
Assuntos
Criopreservação , Glycine max , Lecitinas , Nanopartículas , Preservação do Sêmen , Sêmen , Animais , Bovinos , Masculino , Inseminação Artificial , Análise do Sêmen , Motilidade dos Espermatozoides , Espermatozoides , Preservação do Sêmen/métodosRESUMO
Sperm rheotaxis refers to the ability of sperm cells to align their swimming direction with or against fluid flow. Positive rheotaxis (PR) is the tendency of sperm cells to swim against the flow. Herein, we describe sperm rheotaxis in fertile and infertile males, using a microfluidic platform and focus on rheotaxis as a potential marker of male fertility. A previously reported computer-assisted sperm analysis (CASA) plugin for Image-J was used to detect and analyze the motion of human sperm cells in microfluidic environments. The fabricated microchannels mimic the female reproductive tracts and use an image-processing program to monitor sperm swimming behavior in semen samples from fertile and infertile men. We have constructed an image-processing pipeline. The image-processing pipeline incorporated strengthens object detection and particle tracking to adapt to sperm that are out of focus while swimming on the same track. PR% was defined as the number of PR sperm cells over the number of motile sperm cells. The results showed that the percentage of PR correlates with fertility, wherein the fertile male specimens showed a higher PR% than the other groups (P < 0.05). There is no difference in progressive motility between the control group (fertile men with normal sperm analysis) and group 1 (G1; infertile men with normal sperm analysis). However, PR% was lower (P < 0.05) in the G1 group (13.5 ± 0.4%) compared to the control group (40.3 ± 3.3%) and group 2 (G2; infertile with reduced sperm motility) (15.3 ± 4.6%). Thus, PR% may be used as a novel parameter to explain infertility even in situations where basic sperm analysis following the World Health Organization (WHO) guidelines is unable to do so. We propose to use PR% as a novel parameter for sperm analysis and as a method of sperm selection in assisted reproductive technology.
Assuntos
Infertilidade Masculina , Sêmen , Humanos , Masculino , Feminino , Motilidade dos Espermatozoides , Espermatozoides , FertilidadeRESUMO
Herein, the microfluidic device technique was used to investigate the effects of GDF-9 concentrations and exposure time on the ram sperm positive rheotaxis (PR). Semen was collected from six rams and utilized for PR, motility and sperm kinetic parameter analysis using a computer-assisted sperm analysis program with controlled flow velocity following 0, 10, 20 or 30 min of incubation at 37°C with GDF-9 (200 , 400 or 600 ng/ml; semen sample without GDF-9 was used as a control). Results revealed that there was not an interaction between effects of GDF-9 concentrations and incubation duration on PR% (p = .457) and TM% (p = .099). A simple main effects analysis showed that GDF-9 concentrations had an effect on PR% (p = .003). However, the incubation duration did not have an effect on PR% (p = .101). GDF-9 concentrations had not an effect on TM% (p = .817). By contrast, the incubation duration affected on TM% (p = .026). A higher PR% was found (p < .05) at 200 ng GDF-9 after 10 min (46.7 ± 10.3) and 20 min (45.5 ± 11.5) of incubation. After 30 min of incubation, the PR% was found lowest (p < .05) at 400 ng of GDF-9 (30.6 ± 14.1) and 600 ng of GDF-9 (32.2 ± 9.6). There was no difference (p > .05) in the sperm kinetic parameters between the four treatment groups. In conclusion, the ram sperms had the best rheotaxis properties after 10 and 20 min of incubation with 200 ng of GDF-9 and were sensitive to high concretions.
Assuntos
Preservação do Sêmen , Sêmen , Animais , Criopreservação/métodos , Criopreservação/veterinária , Fator 9 de Diferenciação de Crescimento/farmacologia , Masculino , Análise do Sêmen/veterinária , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Ovinos , Carneiro Doméstico , Motilidade dos Espermatozoides , EspermatozoidesRESUMO
COVID-19 is a disease caused by the coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2), known as a highly contagious disease, currently affecting more than 200 countries worldwide. The main feature of SARS-CoV-2 that distinguishes it from other viruses is the speed of transmission combined with higher risk of mortality from acute respiratory distress syndrome (ARDS). People with diabetes mellitus (DM), severe obesity, cardiovascular disease, and hypertension are more likely to get infected and are at a higher risk of mortality from COVID-19. Among elderly patients who are at higher risk of death from COVID-19, 26.8% have DM. Although the reasons for this increased risk are yet to be determined, several factors may contribute to type-2 DM patients' increased susceptibility to infections. A possible factor that may play a role in increasing the risk in people affected by diabetes and/or obesity is the impaired innate and adaptive immune response, characterized by a state of chronic and low-grade inflammation that can lead to abrupt systemic metabolic alteration. SARS patients previously diagnosed with diabetes or hyperglycemia had higher mortality and morbidity rates when compared with patients who were under metabolic control. Similarly, obese individuals are at higher risk of developing complications from SARS-CoV-2. In this review, we will explore the current and evolving insights pertinent to the metabolic impact of coronavirus infections with special attention to the main pathways and mechanisms that are linked to the pathophysiology and treatment of diabetes.
Assuntos
Imunidade Adaptativa , Complicações do Diabetes , Imunidade Inata , Obesidade , Síndrome do Desconforto Respiratório , Fatores Etários , COVID-19/complicações , COVID-19/imunologia , COVID-19/mortalidade , COVID-19/terapia , Complicações do Diabetes/imunologia , Complicações do Diabetes/mortalidade , Complicações do Diabetes/terapia , Diabetes Mellitus Tipo 2/imunologia , Diabetes Mellitus Tipo 2/mortalidade , Diabetes Mellitus Tipo 2/terapia , Humanos , Obesidade/imunologia , Obesidade/mortalidade , Obesidade/terapia , Síndrome do Desconforto Respiratório/etiologia , Síndrome do Desconforto Respiratório/imunologia , Síndrome do Desconforto Respiratório/mortalidade , Síndrome do Desconforto Respiratório/terapia , SARS-CoV-2RESUMO
Rheotaxis of sperm using a microfluidic device was explored in human, mice and bull. However, the rheotaxis of ram sperm and its role in fertility are unknown. Herein, we described the sperm rheotaxis in ram using microfluidic devices and focused on rheotaxis as potential markers of in vivo fertility. Computer-assisted sperm analysis (CASA) with controlled flow velocity was used to explore the kinematic parameters of sperm, total motility and positive rheotaxis (PR). The percentage of PR was defined as the number of PR sperm cells over the number of motile sperm cells. Then, according to the percentage of PR sperm, rams were classified into two groups; sperm with ≥40% PR and <40% PR, although the two ram groups showed similar total motility and kinematic values of sperm evaluated by CASA (p > .05). Two groups of rams mated one hundred thirty ewes naturally (10 ewes/ram). In the results, the pregnancy rate was higher in ≥40% PR (94.4%) than in <40% PR (42.5%, p < .05) after natural mating. Besides, the pregnancy loss was higher in <40% PR (33.3%) than in >40% PR group (8.1%, p < .05). In conclusion, the PR examination in semen can contribute to evaluate the reproductive performance of ram that will provide valuable insights into the semen evaluation.
Assuntos
Fertilidade , Carneiro Doméstico/fisiologia , Motilidade dos Espermatozoides , Animais , Feminino , Processamento de Imagem Assistida por Computador , Masculino , Microfluídica/métodos , Gravidez , Análise do Sêmen/veterináriaRESUMO
In vitro growth (IVG) culture of bovine oocyte-cumulus-granulosa complexes (OCGCs) is generally carried out for 12 or 14 days using conventional gas impermeable culture devices. The culture duration may be longer compared to follicular development in vivo. During follicular development, follicles receive oxygen from micro vessels; however, oxygen supply is limited under the culture using conventional gas impermeable devices. The purpose of this study was to investigate the effect of increasing dissolved oxygen availability using a gas permeable (GP) culture device with or without antioxidant (astaxanthin, Ax) supplementation on 8-day IVG culture systems for bovine OCGCs derived from early antral follicles. We cultured OCGCs in GP, GP supplemented with Ax (GP + Ax), and a conventional gas impermeable device (control) for 8 or 12 days. OCGC viability were significantly higher when cultured for 8 days than 12 days (p < 0.001) in all culture condition, but significant difference was not observed between groups (p > 0.05). Antrum formation rates of OCGCs were higher after 12 days than 8 days of culture in all culture condition (p < 0.001) and were significantly higher in the control than GP groups regardless of Ax supplementation (p < 0.05). Oocyte diameters were similar among day-8 GP + Ax, day-8 control and day-12 control groups (p > 0.05). Nuclear maturation rates of oocytes grown in vitro for 8 days were significantly higher in the GP + Ax group than in the control and the GP groups (p < 0.05) and similar to oocytes grown for 12 days regardless of the culture conditions (p > 0.05). The generation of reactive oxygen species in OCGCs on day 8 of IVG culture was significantly lower in the GP + Ax group than those of the GP and control groups (p < 0.05). IVG oocytes after eight days of culture developed into blastocysts, and the cleavage and blastocyst rates were similar in all treatment groups. However, in vivo-grown oocytes had significantly higher (p < 0.05) cleavage and blastocyst rates than the IVG oocytes in all groups. The present study demonstrates that increased oxygen availability using a GP culture device with Ax supplementation promotes oocyte growth and maturation competence but inhibits proliferation of granulosa cells and antrum formation compared with a conventional gas impermeable culture device, and that OCGCs can attain developmental competence after 8 days of IVG culture.
Assuntos
Oócitos , Oxigênio , Animais , Bovinos , Suplementos Nutricionais , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Folículo Ovariano , XantofilasRESUMO
Improving the quality and the developmental competence of in vitro produced (IVP) embryos is an indispensable goal for assisted reproductive technology. Autophagy is a major protective mechanism for intracellular degradation of unnecessary cytoplasmic components. Autophagy ends by the fusion between autophagic vacuoles and lysosomes, allowing the degradation of the cargo by lysosomal enzymes, especially the cathepsins (CTSs). However, it is still unclear how autophagy and cathepsin K (CTSK) relate to embryo development. This study evaluated (1.) the activities of autophagy and CTSK in relation to bovine embryo quality and (2.) the effect of autophagy induction and/or CTSK inhibition on preimplantation embryo development and quality. We show here that good-quality embryos exhibited a greater autophagic activity and less CTSK activity compared to poor-quality embryos. Blastomeres of an individual embryo may vary in their quality. Good quality blastomeres showed an increased autophagic activity and decreased CTSK activity compared to poor-quality blastomeres within the same embryo at different developmental stages. Importantly, induction of autophagy and/or inhibition of CTSK improved the developmental rate (increased blastocyst and hatching rates) and the quality (increased total cell number and decreased the percentage of apoptotic cells) of IVP bovine embryos. These results demonstrate a promising approach to selectively isolate good-quality embryos and improve the efficiency of IVEP of cattle embryos.
Assuntos
Autofagia/fisiologia , Catepsina K/metabolismo , Desenvolvimento Embrionário/fisiologia , Fertilização in vitro/veterinária , Animais , Bovinos , Técnicas de Cultura Embrionária/veterinária , Embrião de Mamíferos/metabolismo , Feminino , GravidezRESUMO
The aim of the present study was to investigate the effect of kisspeptin-10 (Kp10) injection on semen characteristics, testosterone (T) production and sperm rheotaxis using microfluidic devices in immature ram. Computer-assisted sperm analysis (CASA) with controlled flow velocity was used to explore the kinetic parameters of sperm and positive rheotaxis (PR %). PR % was defined as the number of PR sperms over the number of motile sperms. Healthy Ossimi rams were randomly divided into two groups; a saline-treated control group and Kp10-treated one (5 µg/kg body weight). Treatments were given by intramuscular injection once a week for 1 month. After 1 month, the semen was collected and evaluated weekly for 6 weeks, while the blood samples were collected every 2 weeks for the next 8 weeks. Semen properties were significantly affected by Kp10 injection (p < .01). The Kp10 increased the volume, sperm concentration and percentages of live sperm compared with those of control. Additionally, sperm trajectories and rheotaxis get improved by the injection of Kp10 with time. Furthermore, kisspeptin improved the secretion of testosterone levels throughout the period of study. In conclusion, injections of the Kp10 had a positive impact on semen characteristics as well as improved sperm rheotaxis of Ossimi rams in subtropics.
Assuntos
Kisspeptinas/farmacologia , Análise do Sêmen/veterinária , Espermatozoides/efeitos dos fármacos , Animais , Kisspeptinas/administração & dosagem , Masculino , Carneiro Doméstico , Contagem de Espermatozoides/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacos , Testosterona/sangueRESUMO
Reproductive dysfunction is a common consequence of both obesity and diabetes. This study investigated the impact of obesity and diabetes, alone or combined, on physiological reproductive parameters in male rats. Twenty-four male Wistar Albino rats were divided into four groups: Control; obese non-diabetic; diabetic; and obese diabetic. Obesity was provoked by consumption of a high-fat diet (HFD) consisting of 40% energy from fat for 90 days. Diabetes was induced by an intraperitoneal injection of streptozotocin at a dose of 40 mg/kg/day for three consecutive days. Semen, histopathological, and morphometric analyses were carried out. Serum testosterone, luteinizing hormone (LH), and vaspin and visfatin were measured using ELISA kits. Hypothalamic Kiss-1 mRNA was detected using qPCR and pituitary nitric oxide (NO) was determined using Griess reagent. Our results showed a decrease in semen quality parameters, testosterone, and LH levels with degenerative changes in the testes in experimental groups when compared to control group. This had a positive correlation with hypothalamic Kiss-1 and a negative correlation with pituitary NO and serum vaspin and visfatin. In addition, adverse effects were more pronounced in animals with obesity and diabetes combined compared to rats who were either diabetic or obese. In conclusion, obesity and diabetes, alone or combined, had a negative impact on male rat fertility. Moreover, obesity and diabetes combined had more harmful effects on male fertility when compared with obesity alone. Hypothalamic Kiss-1, pituitary NO, and serum vaspin and visfatin may play a role in the pathophysiology of male infertility-associated with obesity and diabetes.
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The antral follicle count (AFC) is used as an indicator of cow fertility. We herein investigated the relationship between AFC and the steroidogenesis of granulosa cells and confirmed the developmental competence of oocytes derived from early antral follicles (0.5-1.0 mm) using in vitro growth culture. Slaughterhouse-derived ovaries were divided into high (≥ 25) and low (< 25) AFC groups based on AFC (≥ 2.0 mm). Oocyte-cumulus-granulosa complexes (OCGCs) collected from early antral follicles were cultured for 12 days. The total number, viability, and diameter of granulosa cells and estradiol-17ß and progesterone production during the culture were evaluated. Surviving oocytes on day 12 were subjected to in vitro maturation, and their volume and nuclear status were evaluated. Some oocytes were subjected to the evaluation of developmental competence to blastocysts. Although the total number and viability of granulosa cells did not differ between the groups, granulosa cell diameters were smaller in the high AFC group than in the low AFC group. The estradiol-17ß and progesterone ratio on day 8 was higher in the high AFC group than in the low AFC group. Oocyte volumes and nuclear maturation rates were greater in the high AFC group than in the low AFC group. The development rate to blastocysts was 9.1% in the high AFC group, while no oocytes developed to blastocysts in the low AFC group. Therefore, estradiol-17ß production by granulosa cells appears to be greater in high AFC cattle than in low AFC cattle, thereby promoting the acquisition of oocyte competence.
Assuntos
Células do Cúmulo/citologia , Células da Granulosa/metabolismo , Oócitos/citologia , Folículo Ovariano/metabolismo , Animais , Bovinos , Células Cultivadas , Estradiol/metabolismo , Feminino , Células da Granulosa/citologia , Folículo Ovariano/citologia , Progesterona/metabolismoRESUMO
We investigated the effects of pre-maturational (pre-IVM) culture on the developmental competence of small-sized bovine oocytes (110 and < 115 µm). Oocytes were cultured with 3-isobutyl-1-methylxanthine (IBMX) for 0, 5, or 10 h and subjected to in vitro maturation, fertilization, and culture. The cleavage rate (73%) of small-sized oocytes with 5 h pre-IVM was higher than those with 0 and 10 h pre-IVM (61 and 62%, respectively). The blastocyst rate (16%) of embryos derived from small-sized oocytes with 5 h pre-IVM was higher than those with 0 and 10 h pre-IVM (9 and 8%, respectively). In addition, small-sized oocytes with 5 h pre-IVM had a higher mean cell number in blastocysts (134.1 ± 34.8) than those with 0 and 10 h pre-IVM (100.2 ± 17.2 and 107.8 ± 23.7, respectively). In conclusion, the pre-IVM of small-sized oocytes with IBMX for 5 h improved the developmental competence of bovine oocytes, as well as the quality of blastocysts.
Assuntos
Blastocisto/efeitos dos fármacos , Fertilização in vitro/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/crescimento & desenvolvimento , 1-Metil-3-Isobutilxantina/farmacologia , Animais , Bovinos , Feminino , Oócitos/efeitos dos fármacosRESUMO
There is an increasing concern over male reproductive toxicity caused by lead exposure. Folic acid (FA) is supposed to be a promising therapeutic strategy against lead toxicity. Therefore, the aim of this experimental study was to shed light on the potential protective role of FA on lead-induced testicular dysfunction in rats and its possible underlying mechanistic pathways. Rats (n = 24) were divided into four groups: Control, FA, Lead, and FA+Lead group. After 4 weeks, lead intoxication resulted in a marked reduction in the relative testicular weight and the serum level of testosterone, an impairment in the characters of semen analysis, and an increased content of lead, malondialdehyde and both interleukin-6 and -10 and a decreased antioxidant enzyme levels in the testicular tissue homogenate. Furthermore, marked degenerative histological changes and an increased expression of NF-κB were also noticed in the testicular tissue of Lead group. Supplementation of FA in association with lead considerably alleviated these adverse outcome responses most probably owing to its cytoprotective ability as emerged from combating the oxidative stress and inflammatory reactions. We concluded that FA could act as a highly effective fighting approach against lead-associated testicular toxicity.
Assuntos
Ácido Fólico/administração & dosagem , Intoxicação por Chumbo/tratamento farmacológico , Intoxicação por Chumbo/fisiopatologia , Doenças Testiculares/prevenção & controle , Doenças Testiculares/fisiopatologia , Testosterona/sangue , Animais , Citoproteção/efeitos dos fármacos , Intoxicação por Chumbo/patologia , Masculino , Ratos , Ratos Wistar , Análise do Sêmen , Espermatozoides/efeitos dos fármacos , Espermatozoides/patologia , Doenças Testiculares/induzido quimicamente , Resultado do TratamentoRESUMO
The present study investigated the effects of bovine granulosa cell monolayers (BGML) and canine granulosa cell monolayers (CGML) on nuclear maturation of canine oocytes with and without cumulus cells. Cumulus-oocyte complexes (COCs) or cumulus-free oocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM, control group), DMEM with BGML (BGML group), or DMEM with CGML (CGML group) for 72 h at 38.5 °C in 5% CO(2), 5% O(2,) and 90% N(2). All media were supplemented with 10% of FCS, 50 ng/mL of EGF, 2 µg/mL of estradiol-17ß, 0.1 IU/mL of hCG, 0.1 IU/mL of FSH, 0.25 mM of pyruvic acid, 100 µM of ß-mercaptoethanol, 100 IU/mL of penicillin, and 100 µg/mL of streptomycin. In cumulus-enclosed oocytes retrieved from ovaries at estrus and/or diestrus, the highest percentage of M-II oocytes (P < 0.05) was present in the BGML group (27.0%) compared with the CGML group (7.9%) and the control group (3.5%). In cumulus-free oocytes collected from ovaries at estrus and/or diestrus, the proportions of M-II oocytes co-cultured with the CGML were low (3.0%) and similar (P > 0.05) to proportions achieved with control (3.0%). However, the presence of BGML improved (P < 0.05) the ability of denuded oocytes to develop into M-II (10.2%). The BGML group had the highest overall meiotic resumption (P < 0.05), and least oocyte degeneration (P < 0.05) among experimental groups. In conclusion, BGML had a positive impact on the in vitro maturation system, as well as meiotic resumption of canine oocytes.
Assuntos
Técnicas de Cocultura , Cães , Células da Granulosa/fisiologia , Oócitos/crescimento & desenvolvimento , Animais , Bovinos , Núcleo Celular/fisiologia , Células Cultivadas , Células do Cúmulo/fisiologia , Feminino , Meiose , Oócitos/ultraestruturaRESUMO
The assisted reproductive techniques (ARTs) such as in vitro fertilization, embryo transfer, and cryopreservation of gametes have contributed considerably to the development of biomedical sciences in addition to improving infertility treatments in humans as well as the breeding of domestic animals. However, ARTs used in canine species have strictly limited utility when compared with other mammalian species, including humans. Although successful somatic cell cloning has been reported, artificial insemination by frozen semen to date is only available for the improved breeding and reproduction for companion and working dogs as well as guide dogs for the blind. We describe here the successful cryopreservation of embryos and subsequent embryo transfer in dogs. Canine embryos were collected from excised reproductive organs after artificial insemination and subsequently cryopreserved by a vitrification method. When the 4-cell to morula stage of cryopreserved embryos were nonsurgically transferred into the uteri of nine recipient bitches using a cystoscope, five recipients became pregnant and four of them delivered a total of seven pups. The cryopreservation of embryos in canine species will facilitate the transportation and storage of genetic materials and will aid in the elimination of vertically transmitted diseases in dogs. In addition, this technique will contribute to the improved breeding of companion and working dogs such as guide dogs, drug-detecting dogs, and quarantine dogs.
Assuntos
Criopreservação/veterinária , Cães/embriologia , Transferência Embrionária/veterinária , Animais , Cruzamento/métodos , Transferência Embrionária/instrumentação , Transferência Embrionária/métodos , Feminino , Inseminação Artificial , Mórula , GravidezRESUMO
PURPOSE: To investigate the ability of medium conditioned with bovine cumulus-oocyte complexes (COCs) to support nuclear maturation of canine oocytes recovered from domestic dog ovaries. METHODS: Cumulus-oocyte complexes were obtained from ovaries of domestic bitches (8 months old to 7 years old), and in-vitro maturation was evaluated in TCM-199 supplemented with different concentrations (0, 20, 30 or 50%) of bovine COCs-conditioned medium (BCM). The canine COCs were cultured for 72 or 96 h at 38.5°C in 5% CO2, 5% O2 and 90% N2. The bovine COCs-conditioned medium was obtained from culture of bovine COCs with TCM-199 supplemented with 5% FCS for 22 h at 38.5°C in 2% CO2, 98% air. RESULTS: The proportion of germinal vesicle breakdown (GVBD) after 72 h was significantly higher (P < 0.05) in medium supplemented with 30% BCM (20.7%) compared with the control group (13.4%). The rates of GVBD-MII stage were significantly higher (P < 0.05) when oocytes were matured with BCM at concentration of 30% (41.5%) compared with control (26.6%) after 72 h in-vitro culture. After 96 h in-vitro culture, the oocytes matured in medium supplemented with 30% BCM (5.5%) showed a significant increase (P < 0.05) in the proportion of MII compared with control (0.7%). However, increasing the cultivation time from 72 to 96 h resulted in an increase in oocyte degeneration rate. CONCLUSIONS: The results suggested that bovine COCs-conditioned medium supplementation significantly increased nuclear maturation of canine oocytes.
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PURPOSE: To investigate the effect of graft site and gonadotrophins administration on the number and survival rate of follicles of canine ovarian grafts transplanted to NOD-severe combined immune deficiency (SCID) mice. METHODS: Fresh ovarian cortex slices obtained from immature bitches were grafted subcutaneously (SC), under kidney capsule (KC) or into ovarian bursa (OB) in NOD-SCID mice. Two months after surgery, the mice allocated into non-treated and treated gonadotrophins groups that injected with porcine follicle stimulating hormone during 7 days and human chorionic gonadotrophin 48 h later. Ovarian grafts were collected after 10 h of last injection and processed for histology. RESULTS: The number of transitional and preantral follicles under KC and into OB was significantly higher in gonadotrophins-treated mice than those who received saline. Furthermore, the survival rates of primary, transitional and preantral follicles under KC and into OB grafts were significantly higher than those placed SC in the treated gonadotrophins group, and in the non-treated gonadotrophins group; the proportion of primary and preantral follicle survival was significantly higher under KC and into OB than SC grafts. CONCLUSIONS: In canine ovarian xenografting, administration of gonadotrophin could be effective for improvement of survival of transplanted ovary. Furthermore, the grafting into OB appeared to be better than grafting under KC, which in turn is better than SC.
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Epidemiological and clinical data suggest high rates of suicidal behavior in alcohol-dependent individuals. Suicide attempters are likely to be young, to be single or separated, and to have made prior attempts. They differ from non-attempters by higher levels of impulsive aggression, drug use, and psychiatric comorbidity, particularly personality and depressive disorders. Treatment-seeking, alcohol-dependent individuals often present with multiple risk factors. Early recognition of suicidal behavior is hindered, however, by insufficient data regarding the acute phenomenology of imminent risk. Similarly, little research is available to guide intervention efforts. Initial trials support the use of fluoxetine for the treatment of suicidal, alcohol-dependent persons with comorbid depressive disorders. Future studies may clarify the relative efficacy of various psychotherapeutic and pharmacological approaches to treating these patients.
Assuntos
Alcoolismo/epidemiologia , Tentativa de Suicídio/estatística & dados numéricos , Agressão/psicologia , Comorbidade , Transtorno Depressivo/epidemiologia , Fluoxetina/uso terapêutico , Humanos , Comportamento Impulsivo/psicologia , Fatores de Risco , Inibidores Seletivos de Recaptação de Serotonina/uso terapêuticoRESUMO
The superiority of enoxaparin compared with unfractionated heparin in the medical management of patients with non-ST elevation acute coronary syndromes (NSTE ACS) has been demonstrated in clinical trials. Further, enoxaparin has been shown to be safe and effective during PCI, including in combination with glycoprotein IIb/IIIa inhibitors. Whether enoxaparin is superior to unfractionated heparin in patients with NSTE ACS under-going early invasive strategy is currently being tested in a large clinical trial. Data on the use of enoxaparin in patients undergoing primary angioplasty for ST-segment elevation myocardial infarction are limited. Unlike patients who present to the catheterization laboratory after several doses of enoxaparin where in a steady state anticoagulation might have been achieved, patients who present early after administration of a single dose of subcutaneous enoxaparin may not have an adequate level of anticoagulation for PCI. The ability to monitor activity of enoxaparin in such patients using a point-of-care test might be useful. This report describes a patient with ST-segment elevation myocardial infarction who presented for primary angioplasty 75 minutes after administration of subcutaneous enoxaparin. The Rapidpoint Enox test measured 135 seconds and the patient's corresponding serum anti-Xa level was 0.12 IU/mL indicating a suboptimal level of anticoagulation for PCI. Procedural success was attained using additional 0.3-mg/kg intravenous enoxaparin.
Assuntos
Angioplastia Coronária com Balão , Anticoagulantes/uso terapêutico , Eletrocardiografia , Enoxaparina/uso terapêutico , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/terapia , Sistemas Automatizados de Assistência Junto ao Leito , Idoso , Idoso de 80 Anos ou mais , Angiografia Coronária , Humanos , Injeções Subcutâneas , MasculinoRESUMO
Intracoronary beta (beta) radiation decreases the incidence of target lesion revascularization after percutaneous intervention (PCI) for in-stent restenosis (ISR). Cutting balloon (CB) angioplasty may also be superior to other percutaneous techniques for the treatment of ISR. We sought to study the outcomes of patients with ISR who underwent both CB angioplasty and intracoronay beta radiation and compare them to patients with ISR who underwent other PCI techniques without concomitant radiation. We also sought to evaluate the safety and efficacy of pullback intracoronary beta radiation for the treatment of long ISR lesions. Between January 2001 and November 2001, 102 patients (mean age = 55 +/- 13 years) with ISR underwent both CB angioplasty and intracoronay beta radiation. beta radiation was delivered using the Beta Cath (Novoste) 30 mm system, and pullback radiation was performed in 41 patients. A comparison group included a total of 393 patients with ISR who underwent other PCI techniques without concomitant intracoronary radiation therapy. Follow-up was obtained in 99 patients (97%) in the CB angioplasty with intracoronary radiation group and 377 patients (96%) in the comparison group. At follow-up, both target vessel revascularization (TVR) and major adverse cardiovascular events (MACE) occurred significantly less in the CB angioplasty with intracoronary radiation group than in the comparison group (7% vs. 18% for TVR, and 14% vs. 24% for MACE; P < 0.05 for both). In the pullback radiation group, TVR was performed in five patients (12%), and MACE occurred in eight patients (20%). A combination of CB angioplasty and intracoronay beta radiation for ISR seems to yield low rates of subsequent target vessel revascularization and adverse cardiac events. In addition, pullback beta radiation using the Beta Cath (Novoste) 30 mm system is safe and can be used to treat long ISR lesions effectively. Further randomized trials are needed to confirm these findings.