RESUMO

INTRODUCTION: The use of mesenchymal stem cells (MSC) in cytotoxicity tests is an in-vitro alternative model for predicting initial doses. Homeopathic medicines may stimulate the immune system to combat a pathology effectively and have been used for over two centuries. Viscum album (VA) extracts are widely used in the treatment of cancer, due to their immunomodulatory, cytotoxic and pro-apoptotic properties. OBJECTIVE: This study aimed to evaluate the in-vitro growth kinetics of canine MSC in relation to cytotoxicity, cell differentiation and expression of pluripotentiality markers, using a VA preparation at the D1D2 (1×10-1, 1×10-2 potency (VAD1D2). METHODS: MSC were obtained from adipose tissue sampled from a healthy dog that was undergoing an elective veterinary procedure and with its owner's permission. The experiments were performed in three groups: MSC treated with VAD1D2 or diluent or untreated (control). The cytotoxicity was evaluated by MTT assay. The differentiation was induced in three lineages, and apoptotic cell labeling was performed by an Annexin-V test. RESULTS: At the concentration of 10 µL/mL of VA, the number of cells after in-vitro culture was maintained when compared with the control (untreated) group. A significant and gradual decrease in cell viability was recorded as VA concentrations increased. The apoptosis analysis showed that VA at 20 µL/mL presented absolute percentages of initial apoptosis twice as high as at 10 µL/mL, which was similar to the control (untreated group). CONCLUSION: The results suggest that the use of efficient methods to assess the in-vitro cytotoxicity of VA-based homeopathic medicines using MSC lineages may predict the potential action at different concentrations. These findings demonstrated that VAD1D2 interferes with canine MSC growth kinetics.

Assuntos

RESUMO

Mesenchymal stem cells (MSCs) from the umbilical cord (UC) have aroused considerable interest. However, little is known about the maternal effect on these cells. The aim of this study was to verify the impact of the nutritional status of donor goats on the growth and differentiation of MSCs from the UC. At parturition, 19 goats were grouped based on their low or high body mass index (low BMI, LBMI, n = 9; and high BMI, HBMI, n = 10). UCs were collected during delivery and Wharton's jelly (WJ) fragments cultured. WJ-MSCs were differentiated into osteocytes, adipocytes, chondrocytes, and the population doubling time (PDT) was determined. Samples of WJ-MSCs were also used to verify the expression of the CD90, CD73, CD34, CD45, and CD105 genes. Media used for WJ-MSC primary cultures were analyzed using near-infrared spectroscopy. The lag phase was 7.5 ± 0.6 days and the entire culture took 26.7 ± 1.3 days, with a cell proliferation rate of 8.500 cells/day. The mean PDT from subculture was 30.0 ± 0.7 h. The CD105 gene was sub-expressed in LBMI, and the spectra of the spent media from the second to fourth day of WJ-MSC primary culture were segregated into negative scores by multivariate analysis. We conclude that, in goats, the nutritional balance of the donor did not affect the in vitro growth of MSCs derived from the UC. However, the molecular profile observed in the low BMI group suggests that the use of MSCs for therapeutic purposes should be considered more carefully.

Assuntos

RESUMO

Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.

RESUMO

Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.(AU)

Assuntos

RESUMO

Abstract Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.

RESUMO

Abstract The planning of a new thermal power plant is linked to the various decision elements and evaluation criteria. Factors such as the plant's geographic positioning, primary energy supply points, paths, and means of delivery of this primary energy should be analyzed. Similar studies are imposed when studying the change of a thermoelectric plant's primary energy source occurs. In Brazil, several plants are currently investigating the exchange of their primary fuel from oil to gas due to the decrees issued by ANEEL. This paper presents software, which uses virtual reality to assist in the various stages of the planning process and in the analyses that must be performed. This software was developed for the Hidrotermica Group and had as the primary target the Borborema Thermoelectric Power Plant.

RESUMO

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...(AU)

Assuntos

RESUMO

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...

Assuntos

RESUMO

The aim of this work was to determine the effect of enalapril maleate administration, during oocyte recovery by serial laparoscopic ovum pick-up (LOPU), on the ovarian response and in vitro embryo production (IVP). Twenty cross-bred goats were allocated equally into two groups: Nulliparous and Multiparous. In each group, five animals were selected to receive daily doses of enalapril maleate during the hormonal protocol. Estrus was synchronized by a PGF2α analog, followed 48 h later by insertion of an intravaginal device with progesterone. Forty-eight hours after, a single dose of FSH/eCG was administered. The FSH/eCG doses were repeated three times, on every four day. Oocytes were recovered by LOPU 24 h after each FSH/eCG dose. Viable oocytes were matured in vitro, to be parthenogenetically activated and cultured for 72 h to the cleavage stage. The drug treatment increased the proportion of total follicles observed at LOPU (p < 0.01) in multiparous goats. In both parity groups, enalapril administration had no effect on the proportion or quality of oocytes recovered. Furthermore, the number of embryos cleaved was similar between the groups. Thus, enalapril maleate affected the ovarian response in multiparous animals only and had no effect on the oocyte quality or IVP.

RESUMO

The aim of this study was to verify the effect of the energy source for a short-term diet supplementation on follicular dynamics, ovarian response and oocyte recovery in goats. Thirty Anglo Nubian crossbred does received a diet for 4 weeks to satisfy the nutritional requirements of breeding for adult non-dairy goats. Seven days prior to oocyte recovery (OR), a group of does (n = 10) was supplemented with ground full-fat linseed in the diet (Diet A), whereas a second group of does (n = 10) received crude glycerine in the diet (Diet B). The total mixed ration (TMR) diet was maintained as the Control Diet (n = 10). All animals were oestrous-synchronized by the use of a progesterone insert for 12 days prior to OR. Follicles were stimulated by using pFSH (five 40-mg/ml doses) during the supplementation time. At OR, follicles were counted and recovered oocytes were classified as viable or degenerated. Follicular dynamics was monitored by ultrasonography, and plasma glucose, cholesterol and triglyceride levels were measured during supplementation. Glucose was higher in Diet B and cholesterol in Diet A. Diet B had a lower proportion of small (<3 mm) and large follicles (≥3 mm; p = 0.01). The follicular growth rate was higher in Diet A (p < 0.01), with follicles emerging in the 5th day of supplementation. No differences were observed for follicles counted and oocytes recovered. Thus, the type of energy source supplemented for a short term was capable to alter the follicular dynamics, without affecting the proportion of morphologically viable oocytes upon recovery.

Assuntos

RESUMO

O objetivo deste trabalho foi desenvolver um modelo preditivo através de técnica multivariada para diferenciar meios de cultivo de células-tronco cultivadas in vitro e criopreservadas de acordo com os perfis de absorbância obtidas por NIR. Para tanto, foram coletados meios de cultivo de células-tronco oriundo do fluido amniótico de fetos caprinos, antes e após o processo de criopreservação por vitrificarão, e submetidos à análise pelo NIR. Foi possível estimar com alta acurácia o tratamento empregado nas amostras, gerando uma impressão digital dos meios de cultivo in vitro de células criopreservadas ou não.(AU)

The objective of this work was to develop a predictive model through a multivariate technique to differentiate culture media from stem cells cultured in vitro and cryopreserved according to the absorbance profiles obtained by NIR. For this purpose, culture media were collected from stem cells from the amniotic fluid of goat fetuses, before and after cryopreservation by vitrification, and submitted to NIR analysis. It was possible to estimate with high accuracy the treatment used in the samples, generating a fingerprint of in vitro culture media of cryopreserved cells or not.(AU)

Assuntos

RESUMO

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.(AU)

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers. (AU)

Assuntos

RESUMO

O objetivo do estudo foi avaliar a viabilidade e taxa de proliferação de células-tronco mesenquimais (CTMs) derivadas do líquido amniótico (LA) após cultivo in vitro, e o efeito de dois agentes crioprotetores. Foram utilizadas 9 cabras prenhes. As amostras foram colhidas de fetos caprinos por laparotomia, e delas obtidos as (CTMs) e submetidas ao cultivo in vitro. Posteriormente, uma fração das células foi criopreservada em meio DMSO (dimetilsulfóxido) ou glicerol e vitrificadas para posterior avaliação da viabilidade. O meio DMSO promoveu melhores taxas de sobrevida celular preservando as características de pluripotencialidade e de replicação in vitro.(AU)

The objective of the study was to evaluate the viability and proliferation rate of mesenchymal stem cells (MTCs) derived from amniotic fluid (LA) after in vitro culture, and the effect of two cryoprotective agents. 9 pregnant goats were used. The samples were collected from goat fetuses by laparotomy, and from them (CTMs) were obtained and cultured in vitro. Subsequently, a fraction of the cells were cryopreserved in DMSO (dimethylsulfoxide) or glycerol medium and vitrified for further evaluation of viability. The DMSO medium promoted better cell survival rates while preserving the characteristics of pluripotency and replication in vitro.(AU)

Assuntos

RESUMO

O objetivo do trabalho foi avaliar o efeito do escore da condição corporal (ECC) materno sobre a capacidade de isolamento, expansão e caracterização de células-tronco mesenquimais (CTMs) derivadas do cordão umbilical de fetos caprinos. Para tanto, dezenove cabras mestiças adultas, pluríparas foram agrupadas de acordo com o ECC, atribuindo-se um escore de 1-5, (GB) grupo baixo com menor ECC, (2,3±0,1, GB, n = 9) e (GA) grupo alto com maior ECC (2,9±0,1, GA, n = 10). Durante o parto, fragmentos do cordão umbilical foram coletados e cultivados in vitro e avaliados a taxa de proliferação celular. Nenhum efeito significativo do ECC foi encontrado para os parâmetros considerados.(AU)

The objective of this study was to evaluate the effect of the maternal body condition score (ECC) on the ability to isolate, expand and characterize mesenchymal stem cells (MTCs) derived from the umbilical cord of goat fetuses. Nineteen crossbred adult crossbred goats were grouped according to ECC, giving a score of 1-5 (GB) low group with lower ECC, (2.3±0.1, GB, n = 9 ) and (GA) high group with higher ECC (2.9±0.1, GA, n = 10). During delivery, umbilical cord fragments were collected and cultured in vitro and the rate of cell proliferation was evaluated. No significant effect of ECC was found for the parameters considered.(AU)

Assuntos

RESUMO

O objetivo deste trabalho foi desenvolver um modelo preditivo através de técnica multivariada para diferenciar meios de cultivo de células-tronco cultivadas in vitro e criopreservadas de acordo com os perfis de absorbância obtidas por NIR. Para tanto, foram coletados meios de cultivo de células-tronco oriundo do fluido amniótico de fetos caprinos, antes e após o processo de criopreservação por vitrificarão, e submetidos à análise pelo NIR. Foi possível estimar com alta acurácia o tratamento empregado nas amostras, gerando uma impressão digital dos meios de cultivo in vitro de células criopreservadas ou não.

The objective of this work was to develop a predictive model through a multivariate technique to differentiate culture media from stem cells cultured in vitro and cryopreserved according to the absorbance profiles obtained by NIR. For this purpose, culture media were collected from stem cells from the amniotic fluid of goat fetuses, before and after cryopreservation by vitrification, and submitted to NIR analysis. It was possible to estimate with high accuracy the treatment used in the samples, generating a fingerprint of in vitro culture media of cryopreserved cells or not.

Assuntos

RESUMO

O objetivo do estudo foi avaliar a viabilidade e taxa de proliferação de células-tronco mesenquimais (CTMs) derivadas do líquido amniótico (LA) após cultivo in vitro, e o efeito de dois agentes crioprotetores. Foram utilizadas 9 cabras prenhes. As amostras foram colhidas de fetos caprinos por laparotomia, e delas obtidos as (CTMs) e submetidas ao cultivo in vitro. Posteriormente, uma fração das células foi criopreservada em meio DMSO (dimetilsulfóxido) ou glicerol e vitrificadas para posterior avaliação da viabilidade. O meio DMSO promoveu melhores taxas de sobrevida celular preservando as características de pluripotencialidade e de replicação in vitro.

The objective of the study was to evaluate the viability and proliferation rate of mesenchymal stem cells (MTCs) derived from amniotic fluid (LA) after in vitro culture, and the effect of two cryoprotective agents. 9 pregnant goats were used. The samples were collected from goat fetuses by laparotomy, and from them (CTMs) were obtained and cultured in vitro. Subsequently, a fraction of the cells were cryopreserved in DMSO (dimethylsulfoxide) or glycerol medium and vitrified for further evaluation of viability. The DMSO medium promoted better cell survival rates while preserving the characteristics of pluripotency and replication in vitro.

Assuntos

RESUMO

O objetivo do trabalho foi avaliar o efeito do escore da condição corporal (ECC) materno sobre a capacidade de isolamento, expansão e caracterização de células-tronco mesenquimais (CTMs) derivadas do cordão umbilical de fetos caprinos. Para tanto, dezenove cabras mestiças adultas, pluríparas foram agrupadas de acordo com o ECC, atribuindo-se um escore de 1-5, (GB) grupo baixo com menor ECC, (2,3±0,1, GB, n = 9) e (GA) grupo alto com maior ECC (2,9±0,1, GA, n = 10). Durante o parto, fragmentos do cordão umbilical foram coletados e cultivados in vitro e avaliados a taxa de proliferação celular. Nenhum efeito significativo do ECC foi encontrado para os parâmetros considerados.

The objective of this study was to evaluate the effect of the maternal body condition score (ECC) on the ability to isolate, expand and characterize mesenchymal stem cells (MTCs) derived from the umbilical cord of goat fetuses. Nineteen crossbred adult crossbred goats were grouped according to ECC, giving a score of 1-5 (GB) low group with lower ECC, (2.3±0.1, GB, n = 9 ) and (GA) high group with higher ECC (2.9±0.1, GA, n = 10). During delivery, umbilical cord fragments were collected and cultured in vitro and the rate of cell proliferation was evaluated. No significant effect of ECC was found for the parameters considered.

Assuntos

RESUMO

Na última década houve um crescente interesse na investigação quanto à presença de células-tronco no fluido amniótico, devido a facilidade de obtenção, isolamento e cultivo in vitro. Além disso, a utilização do fluido como fonte de células-tronco possibilita a obtenção de células com alto grau de indiferenciação e elevada taxa de proliferação in vitro e grande capacidade de diferenciação. Todas estas vantagens tornam o líquido amniótico uma fonte atrativa de células-tronco para utilização em biotecnologias reprodutivas, como a clonagem e transgênese, bem como ensaios clínicos e terapias celulares em animais para posterior utilização em seres humanos. Este artigo apresenta um panorama da pesquisa científica com células-tronco derivadas do fluido amniótico no mundo, a partir de levantamento bibliográfico de artigos científicos de pesquisadores brasileiros e estrangeiros.

In the last decade there has been a growing interest in investigation of presence of stem cells in amniotic fluid due to the ease of obtaining, isolating and in vitro culture. In addition, the use of amniotic fluid as a source of stem cells makes it possible to obtain cells with a high degree of indifferentiation and a high rate of in vitro proliferation and a great capacity for differentiation. All of these advantages make amniotic fluid an attractive source of stem cells for use in reproductive biotechnologies, such as cloning and transgenesis, as well as clinical trials in animals for further use in humans. This article presents a panorama of the scientific research with stem cells derived from amniotic fluid in the world, from a bibliographical survey of scientific articles of Brazilian and foreign researchers.

Assuntos

RESUMO

This study was conducted to evaluate the effects of different feeding levels on the proteome of oviduct and uterus tissues of hormonally stimulated goats during the periovulatory period. Forty goats were separated into four different diet groups: Diet 1.0 M (n = 11), Diet 1.3 M (n = 10), Diet 1.6 M (n = 9), Diet 1.9 M (n = 10), fed with 1.0, 1.3, 1.6 and 1.9 times live weight maintenance, respectively. After four weeks of treatment, six hormonally stimulated females per treatment group were randomly selected for collection of uterine and the oviduct tissue samples. Samples were collected after animals were slaughtered in a commercial unit. Feeding goats with 1.3 to 1.9 times more nutrients than a control group directly influenced the proteome of the oviduct and uterus, altering the expression of proteins that participate in biological processes such as apoptosis, antioxidant, and immunological activities. These events are crucial for fertilization and early embryonic survival. Expression of oviduct proteins such as Tubulin Beta 2B, Transferrin and Disulphide-isomerase A3 increased in the 1.9 M group in relation to the other feeding levels. Disulphide-isomerase A4 showed higher expression in the 1.0 M group compared to diets with higher energetic levels. As energy intake increased in the diets, there was higher expression of Alpha-1-antitrypsin and downregulation of Profilin-1 in the uterus of the goats. In conclusion, this study showed that specific proteins of the goat oviduct and uterus expressed during the periovulatory period are modified as the result of nutritional balance.

Assuntos

RESUMO

The aim of this study was to determine the effects of the cumulative gain in expertise in carrying out handmade cloning (HMC) procedures on embryo yield and pregnancy outcome in cattle. Results from in vitro and in vivo embryo development after HMC during three periods of 7 months, separated by 3-month intervals, were compiled and designated as P1, P2 and P3. Blastocyst yield, morphological quality and stage of development, and pregnancy per embryo transfer (ET) on Day 30 of gestation were compared. Zona-intact oocytes were activated chemically in each experiment replicate, and development of parthenogenetic blastocysts was used as a control measurement of oocyte quality and in vitro culture conditions. A total of 21,231 cumulus-oocyte complexes (COCs) were in vitro-matured, with 5,432, 10,721 and 5078 COCs used in 16, 18 and 10 replicates for P1, P2 and P3, respectively. Cloned blastocyst yields on Day 7 increased from 15.5% (124/798) in P1 to 21.6% (309/1428) and 36.6% (280/764) in P2 and P3, respectively. No differences were observed in blastocyst development of parthenogenetic embryos, which average 30.0, 37.6, and 36.4% in P1, P2, and P3, respectively. A 10-fold higher probability of obtaining cloned blastocysts at more advanced stages of development and of higher morphological grade was seen during P3 compared with P1. Pregnancy per ET on Day 30 also increased with gain in expertise, being 6.7% (2/30), 20.8% (10/48) and 40.0% (24/60) for P1, P2 and P3, respectively. The relative efficiency for the establishment of pregnancies (per total COC) increased from 0.04% (1:2716) in P1 to 0.22% (1:460) in P2, reaching 0.47% (1:212) in P3. Results demonstrated a gradual improvement in in vitro and in vivo embryo development over time after establishment of HMC procedures in the laboratory, highlighting the importance of gaining experience and technical skills on the overall cloning efficiency.

Assuntos