This document summarizes the process of obtaining farm animal embryos in vitro, which consists of three main steps: in vitro maturation of oocytes, in vitro fertilization of matured oocytes, and in vitro culture of embryos. It describes the techniques used for each step, differences among farm animal species, and challenges faced, such as polyspermy in pigs and horses. The overall goal of this procedure is to produce offspring using assisted reproduction techniques for applications in animal breeding, research, and treatment of infertility.
Basic Formal Ontology (BFO) and DiseaseBarry Smith
The document discusses different approaches to conceptualizing health, disease, and biological kinds across multiple levels of granularity. It notes that traditional biology data conceptualized entities based on observable instances, while new biology data represents entities at the molecular level through genetic sequences. It argues that linking different kinds of phenomena represented at various levels requires annotation with terms from controlled vocabularies like ontologies. Ontologies provide a structured framework for integrating data across databases and supporting logical reasoning by standardizing references to biological entities, processes, and functions.
This document is a curriculum vitae for Lei Li, a professor at the State Key Laboratory of Stem Cell and Reproductive Biology at the Chinese Academy of Sciences. It outlines her education, degrees, professional experience, awards, publications, and invited presentations. She received her PhD from Gunma University in Japan and has held positions at several institutions in China, Japan, and the United States. Her research focuses on early mouse embryonic development and diabetes.
The mammalian-specific Tex19.1 gene plays an essential role in spermatogenesi...Yara Tarabay
The mammalian-specific Tex19.1 gene plays an important role in spermatogenesis and placenta-supported development in mice. Deletion of the Tex19.1 gene in mice leads to impaired spermatogenesis and male sterility. It also causes perinatal lethality due to placental defects, resulting in severe hypotrophy in newborn homozygous mutant mice regardless of sex. Furthermore, deletion of Tex19.1 results in overexpression of retrotransposons, including MMERVK10C, in embryonic stem cells and testes. This suggests Tex19.1 plays a role in controlling retrotransposon expression and has dual functions in male fertility and placental development.
Maternal curcumin exposure causes fetal gross morphological anomalies and ske...Innspub Net
Curcumin is a phenolic compound extracted from the rhizome of turmeric (Curcuma longa L.). Although declared as safe for human consumption, curcumin has been found to be embryotoxic in some organisms indicating its potential as a teratogen. In this study, the teratogenic effect of maternal curcumin exposure in mouse fetuses was evaluated. Three experimental groups of pregnant mice were treated with 1.05, 1.52, and 2.0 mg/g body weight/day 95% curcumin, respectively, from gestation day (GD) 6 to 15. A fourth group without curcumin exposure served as a control. At GD18, the mice were sacrificed and the total number of implanted embryos including resorbed, dead, and live fetuses were counted for litter analysis. Extracted fetuses were also analyzed for gross morphological anomalies and subsequently have undergone alizarin staining for the visualization of skeletal malformations. Results showed an increased resorption rate in the 2.0 mg/g treatment (p<0.001). There is also a reduction of fetal weight (p<0.001) and crown-rump length (p<0.001) in a dose-dependent manner. Gross morphological analysis shows cranio-facial malformations such as flattened nose bridge (p<0.05) and micrognathia (p<0.05) in 2.0 mg/g treatment. Skeletal malformations such as large anterior fontanelle (p<0.001), misaligned ossification centers in the sternum (p<0.001), and delayed ossification in the forepaws, hind paws, and caudal vertebrae (p<0.001) were also observed at 2.0 mg/g treatment. Meanwhile, the presence of supernumerary ribs is not statistically different in the four groups. The results indicate that curcumin is teratogenic in mouse fetuses due to observed gross morphological anomalies and skeletal malformations.
This document analyzes 2,204 human oocytes and embryos from fertilization through the blastocyst stage using microarray comparative genomic hybridization to determine chromosome abnormalities. It finds that aneuploidy rates increase dramatically with female age and that complex abnormalities are common. Chromosome errors originate from failures in meiotic cell division and early mitosis. Most aneuploid embryos survive until the blastocyst stage but likely fail to implant, indicating selection against aneuploidy occurs late in preimplantation development.
This study compared pregnancy outcomes using fresh versus vitrified and thawed oocytes using a novel vitrification method called Vitri-Inga. The study analyzed 125 IVF procedures, with 79 using fresh oocytes and 46 using vitrified-thawed oocytes. Results found no statistically significant differences in fertilization rates, pregnancy rates, or implantation rates between the fresh and vitrified groups. However, the average number of blastomeres was slightly higher in the fresh oocyte group. The study concluded that the Vitri-Inga method effectively preserves human oocyte potential for fertilization and development.
This document discusses proteomics research for biomarkers of endometriosis. It provides the following key points:
1) Proteomics studies the expressed proteins in cells/tissues and can detect protein changes that genomic methods may miss, with the goal of finding biomarkers for endometriosis diagnosis.
2) Despite many studies, no single biomarker or panel has proven accurate enough for clinical use due to the heterogeneous nature of endometrial tissue.
3) The document reviews various biological samples and mass spectrometry techniques used in proteomics research for endometriosis, including plasma, peritoneal fluid, endometrial fluid and tissue.
Basic Formal Ontology (BFO) and DiseaseBarry Smith
The document discusses different approaches to conceptualizing health, disease, and biological kinds across multiple levels of granularity. It notes that traditional biology data conceptualized entities based on observable instances, while new biology data represents entities at the molecular level through genetic sequences. It argues that linking different kinds of phenomena represented at various levels requires annotation with terms from controlled vocabularies like ontologies. Ontologies provide a structured framework for integrating data across databases and supporting logical reasoning by standardizing references to biological entities, processes, and functions.
This document is a curriculum vitae for Lei Li, a professor at the State Key Laboratory of Stem Cell and Reproductive Biology at the Chinese Academy of Sciences. It outlines her education, degrees, professional experience, awards, publications, and invited presentations. She received her PhD from Gunma University in Japan and has held positions at several institutions in China, Japan, and the United States. Her research focuses on early mouse embryonic development and diabetes.
The mammalian-specific Tex19.1 gene plays an essential role in spermatogenesi...Yara Tarabay
The mammalian-specific Tex19.1 gene plays an important role in spermatogenesis and placenta-supported development in mice. Deletion of the Tex19.1 gene in mice leads to impaired spermatogenesis and male sterility. It also causes perinatal lethality due to placental defects, resulting in severe hypotrophy in newborn homozygous mutant mice regardless of sex. Furthermore, deletion of Tex19.1 results in overexpression of retrotransposons, including MMERVK10C, in embryonic stem cells and testes. This suggests Tex19.1 plays a role in controlling retrotransposon expression and has dual functions in male fertility and placental development.
Maternal curcumin exposure causes fetal gross morphological anomalies and ske...Innspub Net
Curcumin is a phenolic compound extracted from the rhizome of turmeric (Curcuma longa L.). Although declared as safe for human consumption, curcumin has been found to be embryotoxic in some organisms indicating its potential as a teratogen. In this study, the teratogenic effect of maternal curcumin exposure in mouse fetuses was evaluated. Three experimental groups of pregnant mice were treated with 1.05, 1.52, and 2.0 mg/g body weight/day 95% curcumin, respectively, from gestation day (GD) 6 to 15. A fourth group without curcumin exposure served as a control. At GD18, the mice were sacrificed and the total number of implanted embryos including resorbed, dead, and live fetuses were counted for litter analysis. Extracted fetuses were also analyzed for gross morphological anomalies and subsequently have undergone alizarin staining for the visualization of skeletal malformations. Results showed an increased resorption rate in the 2.0 mg/g treatment (p<0.001). There is also a reduction of fetal weight (p<0.001) and crown-rump length (p<0.001) in a dose-dependent manner. Gross morphological analysis shows cranio-facial malformations such as flattened nose bridge (p<0.05) and micrognathia (p<0.05) in 2.0 mg/g treatment. Skeletal malformations such as large anterior fontanelle (p<0.001), misaligned ossification centers in the sternum (p<0.001), and delayed ossification in the forepaws, hind paws, and caudal vertebrae (p<0.001) were also observed at 2.0 mg/g treatment. Meanwhile, the presence of supernumerary ribs is not statistically different in the four groups. The results indicate that curcumin is teratogenic in mouse fetuses due to observed gross morphological anomalies and skeletal malformations.
This document analyzes 2,204 human oocytes and embryos from fertilization through the blastocyst stage using microarray comparative genomic hybridization to determine chromosome abnormalities. It finds that aneuploidy rates increase dramatically with female age and that complex abnormalities are common. Chromosome errors originate from failures in meiotic cell division and early mitosis. Most aneuploid embryos survive until the blastocyst stage but likely fail to implant, indicating selection against aneuploidy occurs late in preimplantation development.
This study compared pregnancy outcomes using fresh versus vitrified and thawed oocytes using a novel vitrification method called Vitri-Inga. The study analyzed 125 IVF procedures, with 79 using fresh oocytes and 46 using vitrified-thawed oocytes. Results found no statistically significant differences in fertilization rates, pregnancy rates, or implantation rates between the fresh and vitrified groups. However, the average number of blastomeres was slightly higher in the fresh oocyte group. The study concluded that the Vitri-Inga method effectively preserves human oocyte potential for fertilization and development.
This document discusses proteomics research for biomarkers of endometriosis. It provides the following key points:
1) Proteomics studies the expressed proteins in cells/tissues and can detect protein changes that genomic methods may miss, with the goal of finding biomarkers for endometriosis diagnosis.
2) Despite many studies, no single biomarker or panel has proven accurate enough for clinical use due to the heterogeneous nature of endometrial tissue.
3) The document reviews various biological samples and mass spectrometry techniques used in proteomics research for endometriosis, including plasma, peritoneal fluid, endometrial fluid and tissue.
This document discusses using knock-in mouse models to study Huntington's disease. Knock-in models insert the mutant human gene into mice, mimicking the human condition genetically. Researchers created a Hdh (CAG)150 knock-in mouse model that showed age-related neuronal pathology consistent with Huntington's, such as nuclear inclusions and vacuolated cytoplasm. This model has helped identify early pathological events and provides a tool to test therapeutic interventions for Huntington's disease.
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clini...IOSR Journals
Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from csection with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively. Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine, Deoxycholate, PPARG, C/EBPα, etc.
This study analyzed the development of 247 embryos that were transferred as part of 285 IVF treatment cycles. Using an automated time-lapse imaging system, the study monitored embryo development every 15 minutes for 64 hours post fertilization. The study found that certain morphokinetic parameters, such as the timing of cell divisions and intervals between divisions, were significantly correlated with whether or not the embryo successfully implanted. In particular, implantation was predicted by division to 5 cells occurring between 48.8-56.6 hours after fertilization, an interval of less than 0.76 hours between division to 3 and 4 cells, and a cell cycle 2 duration of less than 11.9 hours. Aberrant behaviors like multi-nucleation or uneven blastom
Spermatotoxic impact of bonny light crude oil (BLCO) ingestion on adult male ...lukeman Joseph Ade shittu
Increasing concern has been expressed about the possible declining trend in the sperm quality and sperm count of man as a result of exposure to environmental estrogenic agents in the past few years now. There is a general paucity of knowledge of BLCO ingestion on the reproductive effect. Hence, we aim to evaluate the impact of sub-lethal dose of BLCO ingestion on semen parameters of adult male mice. Initial acute toxicity study was carried out to determine the lethal dose of BLCO, which was calculated to be 37.4 mg/Kg body wt. A sub-lethal dose of 20 mg/Kg bwt /day of BLCO were then given to 8 male mice in the experimental group. While, the control group of 7 animals received equal volume of 0.9% normal saline via oral garvage for 2 weeks. Data were analysed using SPSS 12 statistical software with P < 0.05 considered statistically significant. There was a significant (P < 0.05) weight gain in the treated group with a significant (P < 0.05) reduction in sperm motility in the treated compared with control. The sperm density of treated and control were 14.5 x 106 /ml and 20.5 x 106 /ml respectively. However, there were also no significant difference in the relative testicular weight and sperm density of treated from that of the control respectively. Thus, it was concluded that BLCO ingestion is spermatotoxic in the adult male Swiss mice
Minor Planet Evidence for Water in the Rocky Debris of a Disrupted Extrasolar...Carlos Bella
Evidence has been found for water in the debris of a disrupted extrasolar minor planet orbiting a white dwarf star. Spectroscopic analysis of the white dwarf's atmosphere revealed an excess of oxygen that cannot be explained by oxide minerals alone, indicating the parent body was originally composed of about 26% water by mass. This demonstrates that water-bearing planetesimals can form around higher mass stars that eventually become white dwarfs. The disrupted planetesimal is the source of a circumstellar debris disk closely orbiting the white dwarf remnant.
1) The study examined potential transgenerational effects of developmental exposure to endocrine-disrupting chemicals (EDCs) like bisphenol A, phthalates, and methoxychlor on female rat reproduction.
2) EDC exposure of pregnant F0 rats and their F1 offspring resulted in accelerated puberty in F1 rats for methoxychlor and DES, and delayed puberty for phthalates. DES also reduced F1 litter size.
3) Further, methoxychlor exposure led to irregular estrous cycles in F1 rats. However, these effects were absent in subsequent unexposed generations F2 and F3, indicating a lack of transgenerational effects.
Sexual behavior and fertility of male rats following subacute hemi-orchidectomyInnspub Net
This study was conducted to investigate the sub-acute response of testicular size, body weight, fertility and mating indices in hemi-orchiectomy induced suppression of testosterone levels (<0.5 ng/mL) among adult male Wistar rats (Four months old). Libido, fertility and extended implication of the procedure on F1 generation were also observed. After exposure to relevant experimental procedure, each of the male rats was housed with a proven female breeder to assess mating and fertility indices. Obtained data was statistically analyzed by analysis of variance. Hemi-orchidectomy resulted in compensatory increase in testosterone secretion due to contralateral testicular hypertrophy, mating and fertility indices of hemi-orchidectomized and fully orchidectomized rats were 80%:60% and 20%:0% respectively. The male progeny belonging to F1 generation showed no sign of infertility. The findings suggest that hemi-orchidectomy induces sudden compensatory testicular hypertrophy, immediate Loss of fertility after full orchidectomy, diminished libido rather than complete loss of libido occurs. In conclusion, number of offspring delivery is independent of testosterone level, however testosterone level and its interference with the complex modifications in the hypothalamo-hypophyseal secretions are involved in the central control of male sexual activity.
This document discusses endometrial receptivity and the need for objective biomarkers to diagnose it. While past research identified some potential molecular markers, none have proven clinically useful. Recent transcriptomic analyses using microarrays have defined a gene expression signature that can classify endometrial status, including receptivity, regardless of histological appearance. This led to the development of the Endometrial Receptivity Array (ERA) tool, which uses a customized microarray and algorithm to identify the personalized window of implantation in individual patients, improving outcomes over histological dating alone.
This document summarizes a study that examined the effects of bisphenol A (BPA), bisphenol F (BPF), and high levels of estrogen on oocyte maturation and quality. Mouse oocytes were exposed to varying concentrations of BPA, BPF, and estrogen. The oocytes were then analyzed for spindle formation, centrosome position, and chromosome alignment. Oocytes exposed to BPA, BPF, and high estrogen showed lower maturation rates and more chromosome abnormalities compared to the control group. BPA exposure resulted in more chromosomal errors than BPF exposure. The findings suggest that exposure to synthetic estrogens like BPA and BPF can negatively impact oocyte development and potentially cause birth defects.
Organoids are small 3D tissues that mimic the function of organs. They were first developed in 2007 and have since been used to model many organs including the brain, pancreas, and heart. While organoids are still in the early stages of research, they show promise for medical and scientific uses. Organoids are made by harvesting stem cells from an organ and culturing them in a gel environment where they self-organize into ball-like structures that resemble miniature organs. Researchers hope organoids can be used for drug testing and personalized medicine without the need for animal testing. As techniques for growing organoids continue to advance, so too will their potential applications.
Sesame leaves intake improve and increase epididymal spermatocytes reserve in...lukeman Joseph Ade shittu
Increasing concern has been expressed about the potential effects of both synthetic and natural estrogenic endocrine disruptors (EEDs) on human reproductive health in our environment in the last decade. However, little attention is paid to histomorphometric structural changes of the epididymis. We aim to evaluate the chronic exposure effects of phytoestrogens found in aqueous extract of Sesame radiatum leaves on the male Sprague Dawley (SD) rats’ epididymes. Thirty adult male SD rats were randomly divided into three groups (2 treated and 1 control groups respectively). In the treated groups, a single daily dose of aqueous leaves extract of S. radiatum (14.0 mg/kg and 28.0 mg/kg body weight) were administered via gastric garvage, while, equal volume of normal saline was administered in control group for six weeks. Histomorphometric study of the epididymal tissues and hormonal assay were analyzed using SPSS software and P < 0.05 was considered statistically significant. Significant (P < 0.05) body weight gain in a dose dependent was observed in all the animals. Also, there was significant weight gain in both raw weight and relative organo-somatic weight of the epididymis per 100 g body weight. However, the weight gain was more in the high dose than the low dose group. The epididymal lumen appeared wider and fuller with spermatocytes when compared to the control. There is significant (P > 0.05) increases in testosterone level compared to control, however, the low dose was also significantly lower than the control. Sesame improves the storage capacity for the spermatozoa in the epididymis in a dose related manner.
This study analyzed meiosis in over 100 live human oocytes using high-resolution fluorescence microscopy. The researchers found that:
1) Human oocyte meiosis takes around 16 hours from nuclear envelope breakdown to anaphase onset, which is significantly longer than in other species.
2) Human oocyte spindles form gradually over this 16-hour period in a chromosome-mediated manner, without centrosomes or microtubule organizing centers.
3) This error-prone chromosome-mediated spindle assembly process favors chromosome segregation defects and provides an explanation for high rates of aneuploidy in human eggs.
This study analyzed 522 embryos from 285 couples undergoing ICSI that were transferred and monitored using time-lapse photography every 15 minutes for 64 hours. The study found that several morphokinetic parameters were correlated with embryo implantation, including the timing of cell divisions from 2 to 5 cells and the duration of cell cycles 2 and 3. Embryos displaying abnormal behaviors like multi-nucleation or uneven blastomere size were less likely to implant. The study aims to use these morphokinetic parameters to develop a model to predict embryo implantation potential and improve embryo selection.
Deciphering signaling mechanisms of cartilage tissue engineered alginate scaf...Antonion Korcari
Combination of Systems Biology and Tissue Engineering approaches by creating a mechanism for 3D cartilage phenotype evaluation. More specifically, high-throughput measurements have been used to interrogate intracellular and extracellular activity of 3D cultured chondrocytes, combined with phenotypic measurements of cartilage growth (s-GAG) to correlate the mechanism of cartilage growth, either untreated or treated with different stimuli.
External factors such as culture conditions, stimulation protocols, and patient characteristics can affect the morphokinetic development of human embryos observed through time-lapse monitoring. Specifically, higher FSH doses, higher estrogen levels, underweight BMI, smoking, and PCOS are associated with slower embryo development. Differences in culture media brands and oxygen concentration can also influence timing of developmental stages. While time-lapse monitoring provides more detailed data on embryo development and a tool for quality control, further randomized clinical trials are still needed to determine the impact on pregnancy outcomes.
The use of time lapse photography in an in vitro fertilization programme for ...鋒博 蔡
This study examined the timing of early cell divisions in human embryos using time-lapse imaging. It found that embryos producing high-quality blastocysts and resulting in pregnancies showed uniform timing of cleavage cycles and interphases. Specifically, the second interphase lasted 11±1 hours, the third interphase was 15±1 hours, and the fourth interphase was 23±1 hours. The corresponding cleavage cycles lasted 15±5 minutes, 40±10 minutes, and 55±15 minutes. In contrast, embryos with shortened or prolonged cell cycles showed poor implantation and development. The study also discovered trichotomic mitosis, where embryos cleaved into an abnormal number of cells, in 17% of cases. Only time-
This study examined 1213 human embryos at the cleavage and blastocyst stages to assess the relationship between morphological appearance and chromosomal abnormalities. The researchers found that at the cleavage stage, many embryos with the best morphological scores were still chromosomally abnormal, indicating little effect of aneuploidy on appearance at this stage. However, at the blastocyst stage aneuploidies were less common in embryos with optimal morphology, though some chromosomally abnormal blastocysts still received high scores, particularly for clinically relevant aneuploidies like trisomy 21. While morphology provides some information about chromosomal status at the blastocyst stage, comprehensive chromosome testing remains important for accurate assessment of embryo viability
Software projects that involve cameras are hard for a number of reasons. Here are some key things to watch out for in your next CCTV or IP Camera development project.
This curriculum vitae summarizes the qualifications and experience of Chukwunweike Ezeukwu. He has over 9 years of oil field experience working for companies like BJ Services Nigeria Limited and Mansfield Energy Nigeria Limited. He holds a B.Eng in Petroleum Engineering from University of Benin and works currently as a Senior Completions Engineer at Mansfield Energy, with responsibilities including well completion design, equipment installation and maintenance, and project supervision.
This document discusses using knock-in mouse models to study Huntington's disease. Knock-in models insert the mutant human gene into mice, mimicking the human condition genetically. Researchers created a Hdh (CAG)150 knock-in mouse model that showed age-related neuronal pathology consistent with Huntington's, such as nuclear inclusions and vacuolated cytoplasm. This model has helped identify early pathological events and provides a tool to test therapeutic interventions for Huntington's disease.
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clini...IOSR Journals
Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from csection with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively. Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine, Deoxycholate, PPARG, C/EBPα, etc.
This study analyzed the development of 247 embryos that were transferred as part of 285 IVF treatment cycles. Using an automated time-lapse imaging system, the study monitored embryo development every 15 minutes for 64 hours post fertilization. The study found that certain morphokinetic parameters, such as the timing of cell divisions and intervals between divisions, were significantly correlated with whether or not the embryo successfully implanted. In particular, implantation was predicted by division to 5 cells occurring between 48.8-56.6 hours after fertilization, an interval of less than 0.76 hours between division to 3 and 4 cells, and a cell cycle 2 duration of less than 11.9 hours. Aberrant behaviors like multi-nucleation or uneven blastom
Spermatotoxic impact of bonny light crude oil (BLCO) ingestion on adult male ...lukeman Joseph Ade shittu
Increasing concern has been expressed about the possible declining trend in the sperm quality and sperm count of man as a result of exposure to environmental estrogenic agents in the past few years now. There is a general paucity of knowledge of BLCO ingestion on the reproductive effect. Hence, we aim to evaluate the impact of sub-lethal dose of BLCO ingestion on semen parameters of adult male mice. Initial acute toxicity study was carried out to determine the lethal dose of BLCO, which was calculated to be 37.4 mg/Kg body wt. A sub-lethal dose of 20 mg/Kg bwt /day of BLCO were then given to 8 male mice in the experimental group. While, the control group of 7 animals received equal volume of 0.9% normal saline via oral garvage for 2 weeks. Data were analysed using SPSS 12 statistical software with P < 0.05 considered statistically significant. There was a significant (P < 0.05) weight gain in the treated group with a significant (P < 0.05) reduction in sperm motility in the treated compared with control. The sperm density of treated and control were 14.5 x 106 /ml and 20.5 x 106 /ml respectively. However, there were also no significant difference in the relative testicular weight and sperm density of treated from that of the control respectively. Thus, it was concluded that BLCO ingestion is spermatotoxic in the adult male Swiss mice
Minor Planet Evidence for Water in the Rocky Debris of a Disrupted Extrasolar...Carlos Bella
Evidence has been found for water in the debris of a disrupted extrasolar minor planet orbiting a white dwarf star. Spectroscopic analysis of the white dwarf's atmosphere revealed an excess of oxygen that cannot be explained by oxide minerals alone, indicating the parent body was originally composed of about 26% water by mass. This demonstrates that water-bearing planetesimals can form around higher mass stars that eventually become white dwarfs. The disrupted planetesimal is the source of a circumstellar debris disk closely orbiting the white dwarf remnant.
1) The study examined potential transgenerational effects of developmental exposure to endocrine-disrupting chemicals (EDCs) like bisphenol A, phthalates, and methoxychlor on female rat reproduction.
2) EDC exposure of pregnant F0 rats and their F1 offspring resulted in accelerated puberty in F1 rats for methoxychlor and DES, and delayed puberty for phthalates. DES also reduced F1 litter size.
3) Further, methoxychlor exposure led to irregular estrous cycles in F1 rats. However, these effects were absent in subsequent unexposed generations F2 and F3, indicating a lack of transgenerational effects.
Sexual behavior and fertility of male rats following subacute hemi-orchidectomyInnspub Net
This study was conducted to investigate the sub-acute response of testicular size, body weight, fertility and mating indices in hemi-orchiectomy induced suppression of testosterone levels (<0.5 ng/mL) among adult male Wistar rats (Four months old). Libido, fertility and extended implication of the procedure on F1 generation were also observed. After exposure to relevant experimental procedure, each of the male rats was housed with a proven female breeder to assess mating and fertility indices. Obtained data was statistically analyzed by analysis of variance. Hemi-orchidectomy resulted in compensatory increase in testosterone secretion due to contralateral testicular hypertrophy, mating and fertility indices of hemi-orchidectomized and fully orchidectomized rats were 80%:60% and 20%:0% respectively. The male progeny belonging to F1 generation showed no sign of infertility. The findings suggest that hemi-orchidectomy induces sudden compensatory testicular hypertrophy, immediate Loss of fertility after full orchidectomy, diminished libido rather than complete loss of libido occurs. In conclusion, number of offspring delivery is independent of testosterone level, however testosterone level and its interference with the complex modifications in the hypothalamo-hypophyseal secretions are involved in the central control of male sexual activity.
This document discusses endometrial receptivity and the need for objective biomarkers to diagnose it. While past research identified some potential molecular markers, none have proven clinically useful. Recent transcriptomic analyses using microarrays have defined a gene expression signature that can classify endometrial status, including receptivity, regardless of histological appearance. This led to the development of the Endometrial Receptivity Array (ERA) tool, which uses a customized microarray and algorithm to identify the personalized window of implantation in individual patients, improving outcomes over histological dating alone.
This document summarizes a study that examined the effects of bisphenol A (BPA), bisphenol F (BPF), and high levels of estrogen on oocyte maturation and quality. Mouse oocytes were exposed to varying concentrations of BPA, BPF, and estrogen. The oocytes were then analyzed for spindle formation, centrosome position, and chromosome alignment. Oocytes exposed to BPA, BPF, and high estrogen showed lower maturation rates and more chromosome abnormalities compared to the control group. BPA exposure resulted in more chromosomal errors than BPF exposure. The findings suggest that exposure to synthetic estrogens like BPA and BPF can negatively impact oocyte development and potentially cause birth defects.
Organoids are small 3D tissues that mimic the function of organs. They were first developed in 2007 and have since been used to model many organs including the brain, pancreas, and heart. While organoids are still in the early stages of research, they show promise for medical and scientific uses. Organoids are made by harvesting stem cells from an organ and culturing them in a gel environment where they self-organize into ball-like structures that resemble miniature organs. Researchers hope organoids can be used for drug testing and personalized medicine without the need for animal testing. As techniques for growing organoids continue to advance, so too will their potential applications.
Sesame leaves intake improve and increase epididymal spermatocytes reserve in...lukeman Joseph Ade shittu
Increasing concern has been expressed about the potential effects of both synthetic and natural estrogenic endocrine disruptors (EEDs) on human reproductive health in our environment in the last decade. However, little attention is paid to histomorphometric structural changes of the epididymis. We aim to evaluate the chronic exposure effects of phytoestrogens found in aqueous extract of Sesame radiatum leaves on the male Sprague Dawley (SD) rats’ epididymes. Thirty adult male SD rats were randomly divided into three groups (2 treated and 1 control groups respectively). In the treated groups, a single daily dose of aqueous leaves extract of S. radiatum (14.0 mg/kg and 28.0 mg/kg body weight) were administered via gastric garvage, while, equal volume of normal saline was administered in control group for six weeks. Histomorphometric study of the epididymal tissues and hormonal assay were analyzed using SPSS software and P < 0.05 was considered statistically significant. Significant (P < 0.05) body weight gain in a dose dependent was observed in all the animals. Also, there was significant weight gain in both raw weight and relative organo-somatic weight of the epididymis per 100 g body weight. However, the weight gain was more in the high dose than the low dose group. The epididymal lumen appeared wider and fuller with spermatocytes when compared to the control. There is significant (P > 0.05) increases in testosterone level compared to control, however, the low dose was also significantly lower than the control. Sesame improves the storage capacity for the spermatozoa in the epididymis in a dose related manner.
This study analyzed meiosis in over 100 live human oocytes using high-resolution fluorescence microscopy. The researchers found that:
1) Human oocyte meiosis takes around 16 hours from nuclear envelope breakdown to anaphase onset, which is significantly longer than in other species.
2) Human oocyte spindles form gradually over this 16-hour period in a chromosome-mediated manner, without centrosomes or microtubule organizing centers.
3) This error-prone chromosome-mediated spindle assembly process favors chromosome segregation defects and provides an explanation for high rates of aneuploidy in human eggs.
This study analyzed 522 embryos from 285 couples undergoing ICSI that were transferred and monitored using time-lapse photography every 15 minutes for 64 hours. The study found that several morphokinetic parameters were correlated with embryo implantation, including the timing of cell divisions from 2 to 5 cells and the duration of cell cycles 2 and 3. Embryos displaying abnormal behaviors like multi-nucleation or uneven blastomere size were less likely to implant. The study aims to use these morphokinetic parameters to develop a model to predict embryo implantation potential and improve embryo selection.
Deciphering signaling mechanisms of cartilage tissue engineered alginate scaf...Antonion Korcari
Combination of Systems Biology and Tissue Engineering approaches by creating a mechanism for 3D cartilage phenotype evaluation. More specifically, high-throughput measurements have been used to interrogate intracellular and extracellular activity of 3D cultured chondrocytes, combined with phenotypic measurements of cartilage growth (s-GAG) to correlate the mechanism of cartilage growth, either untreated or treated with different stimuli.
External factors such as culture conditions, stimulation protocols, and patient characteristics can affect the morphokinetic development of human embryos observed through time-lapse monitoring. Specifically, higher FSH doses, higher estrogen levels, underweight BMI, smoking, and PCOS are associated with slower embryo development. Differences in culture media brands and oxygen concentration can also influence timing of developmental stages. While time-lapse monitoring provides more detailed data on embryo development and a tool for quality control, further randomized clinical trials are still needed to determine the impact on pregnancy outcomes.
The use of time lapse photography in an in vitro fertilization programme for ...鋒博 蔡
This study examined the timing of early cell divisions in human embryos using time-lapse imaging. It found that embryos producing high-quality blastocysts and resulting in pregnancies showed uniform timing of cleavage cycles and interphases. Specifically, the second interphase lasted 11±1 hours, the third interphase was 15±1 hours, and the fourth interphase was 23±1 hours. The corresponding cleavage cycles lasted 15±5 minutes, 40±10 minutes, and 55±15 minutes. In contrast, embryos with shortened or prolonged cell cycles showed poor implantation and development. The study also discovered trichotomic mitosis, where embryos cleaved into an abnormal number of cells, in 17% of cases. Only time-
This study examined 1213 human embryos at the cleavage and blastocyst stages to assess the relationship between morphological appearance and chromosomal abnormalities. The researchers found that at the cleavage stage, many embryos with the best morphological scores were still chromosomally abnormal, indicating little effect of aneuploidy on appearance at this stage. However, at the blastocyst stage aneuploidies were less common in embryos with optimal morphology, though some chromosomally abnormal blastocysts still received high scores, particularly for clinically relevant aneuploidies like trisomy 21. While morphology provides some information about chromosomal status at the blastocyst stage, comprehensive chromosome testing remains important for accurate assessment of embryo viability
Software projects that involve cameras are hard for a number of reasons. Here are some key things to watch out for in your next CCTV or IP Camera development project.
This curriculum vitae summarizes the qualifications and experience of Chukwunweike Ezeukwu. He has over 9 years of oil field experience working for companies like BJ Services Nigeria Limited and Mansfield Energy Nigeria Limited. He holds a B.Eng in Petroleum Engineering from University of Benin and works currently as a Senior Completions Engineer at Mansfield Energy, with responsibilities including well completion design, equipment installation and maintenance, and project supervision.
This document discusses the history and future of a camera management software company. It outlines the company's products from 2000-2005 which included live streaming, sharing, and timelapse capabilities. It notes that choosing a business partner is important and mentions needing a business partner, CTO, or sales partner. It asks for help regarding strategic decisions around products, competitors, and finding investors.
This document provides a work plan for an English course offered at Boston International College in Chitwan, Nepal. The course is titled English and is worth 3 credit hours. It will be taught over 42 lecture hours across 15 weeks. The course aims to teach students to read and write effectively using different patterns such as narration, description, process, and argumentation. It will use texts, cases, projects, quizzes and exams for evaluation. Students must maintain 80% attendance and adhere to academic honesty norms.
This document is a user guide for Allen & Heath MixWizard WZ3 audio mixing consoles. It provides instructions on installing and connecting the console, an overview of the front panel layout and features, and specifications. The guide covers warranty information, safety instructions, and details each section of the console including mono and stereo input channels and the master section. It is intended to help users operate the console properly and get the best performance from it.
Experiencias significativas en el preescolar de sordosOscar Barreto
El documento describe las experiencias significativas de los auxiliares sordos Magly Salcedo y Guillermo Ruiz en el preescolar de niños sordos. Enseñan a los niños normas de cortesía, el reconocimiento del lugar, el uso del calendario, los días de la semana, vocabulario sobre animales, hábitos de higiene y alimentación, actividades físicas y el establecimiento de normas. Todo con el objetivo de estimular el lenguaje, pensamiento y desarrollo de los niños.
Thane city has historically had several shallow lakes due to its location and rainfall. However, urbanization has converted many of these lakes into water tanks. There are currently 35 lakes covering an estimated 670,000 square feet. The Thane Municipal Corporation has undertaken efforts to revive these natural resources through cleaning and bioremediation projects starting in 1998 with Kacharali Lake. This involves restoring the ecological balance between microorganisms, plants, and algae to improve water quality. The corporation is now pursuing additional bioremediation and beautification projects for other lakes through partnerships with educational institutions, NGOs, and the private sector. The goal is to restore the lakes while increasing public participation and awareness of conservation.
This document discusses various aspects of disaster response, including evacuation planning and execution. It covers types of evacuation like immediate and pre-warned evacuation. Key aspects of evacuation planning like identifying those who need assistance are outlined. Guidelines for effective evacuation procedures and emergency response are provided. The challenges of mass evacuations are examined through the example of Hurricane Katrina. Considerations for effective disaster response like assessing needs, partnering with local organizations, and distributing aid while promoting independence are also summarized.
1) Interspecies somatic cell nuclear transfer (iSCNT) involves transferring the nucleus of a donor somatic cell from one species into an enucleated egg of another related or unrelated species.
2) While iSCNT embryos from closely related species can develop further than early stages, embryos from more distantly related species generally only develop to the blastocyst stage due to failures in nuclear reprogramming and activation of the donor genome.
3) Studies of iSCNT embryos have provided insights into some of the molecular mechanisms that are incompletely reprogrammed, such as failure to silence donor genes, degrade maternal RNA, and properly activate the donor embryonic genome.
Supplementation with goat follicular fluid in the in vitroAlexander Decker
The document summarizes a study that examined the effects of supplementing goat follicular fluid (GFF) at different levels in an in vitro maturation medium on cumulus expansion and nuclear maturation of goat oocytes. The results showed that cumulus expansion increased with higher levels of GFF in the medium, with the highest levels at 10% GFF. Lower or no GFF in the medium resulted in less cumulus expansion and nuclear maturation, suggesting oocyte needs were not supported without hormones and nutrients from GFF. The study concludes GFF supplementation in maturation medium can improve oocyte development in vitro.
The document discusses using embryonic stem cells and adult stem cells to create artificial gametes and embryos for reproductive medicine. This includes generating germ cells from stem cells and assembling different stem cell types to form embryo-like structures. However, fully replicating human gametogenesis and embryogenesis in vitro faces many challenges, such as optimizing culture systems that mimic the stem cell niche. While assisted reproduction synthetic embryos may help study development and treat infertility, safety concerns remain until protocols can guarantee success in humans as they have in some rodent studies. More research is still needed to realize the potential of this technology.
This study analyzed 2,204 human oocytes, embryos, and blastocysts to investigate the origin and impact of chromosomal abnormalities during early human development. The results showed that aneuploidy rates increased dramatically with female age and many abnormalities were present until the blastocyst stage, suggesting selection against aneuploid embryos occurs late in development. Mechanisms like anaphase lag and congression failure contributed to errors in the first cell divisions. A wide variety of chromosome abnormalities were detected, with implications for understanding the sources of aneuploidy and how they influence embryo viability.
Pregnancy outcome following swim up preparation of both fresh and cryopreserv...lukeman Joseph Ade shittu
This study was designed to assess the impact of swim up preparation of both fresh and cryopreserved sperm on the pregnancy outcome in a private fertility centre in Lagos. A cross-sectional prospective analysis of 34 asthenozoospermic semen samples of men whose wives were undergoing assisted reproduction was studied. The basic semen parameters comprising of the volume, count, and motility of the sperm before and after swim up preparations with pregnancy outcome were measured. For fresh semen (n = 28, mean age = 37.0 ± 1.1 years, mean volume = 2.16 ± 0.1 ml), the sperm count decreased significantly (p<0.01)><0.01)><0.01)><0.01) from 25.1 ± 4.01 to 32.8 ± 6.18%. The pregnancy outcome of cryopreserved was 30%. The pregnancy outcome was higher with fresh than the cryopreserved semen. However, the motility was a significant indicator for the successful outcome. Swim up procedure improve the motility of both cryopreserved and fresh semen with a better pregnancy outcome in this study.
Review Sperm Comparative Morphology and Function Related to Altered Reproductiveanbiocore
This document provides a review of the evolution of sperm structure and function in mammals. It discusses how sperm size, shape, and structures like flagella have diverged among species due to factors like body size, mating systems, and sperm competition. Larger mammals tend to have smaller sperm to produce more of them, while smaller mammals produce fewer but larger sperm. The review also examines how reproductive strategies around fertilization have influenced sperm traits, with species relying more on numbers favoring smaller, faster sperm and those using sperm displacement favoring fewer but more competitive sperm.
Animal Cloning Procedure, Problems and PerspectivesShafqat Khan
Cloning in farm animals has problems and perspectives. Key issues include developmental anomalies in cloned animals, the large offspring syndrome observed in cattle and sheep clones, and safety apprehensions regarding meat and milk from cloned animals. However, cloning also has potential applications for transgenic animal production, creating disease models, bioreactors, and research into xenotransplantation. It allows the propagation of elite livestock and conservation of endangered species. Further optimization is needed to improve cloning efficiency and resolve health issues.
This document discusses building communities around ontology development. It provides examples of gene ontologies, plant ontologies, and trait ontologies that are used to group genes and phenotypes. It also outlines how ontologies are developed, managed, and annotated through collaborations between various organizations. Ontology requests are monitored through bug tracking tools and mailing lists. Participation is encouraged to help drive ontology development and annotation.
This document studied the effects of deer velvet extract from Formosan sika deer on mouse embryonic development and anti-oxidative enzyme expression. Mouse 4-cell embryos were divided into groups and cultured with different concentrations of deer velvet extract or hydrogen peroxide. Embryonic development stages were observed every 12 hours over 72 hours of incubation. The deer velvet extract promoted embryonic development and maintained blastocyst development rates similar to the control when embryos were challenged with hydrogen peroxide. Gene expression of anti-oxidative enzymes in blastocysts was not significantly different between deer velvet treatment groups. The deer velvet extract thus relieved oxidative stress on embryos and supported blastocyst development in vitro.
This document studied the effects of deer velvet extract from Formosan sika deer on mouse embryonic development and anti-oxidative enzyme expression. Mouse 4-cell embryos were divided into groups and cultured with different concentrations of deer velvet extract or hydrogen peroxide. Embryonic development stages were observed every 12 hours over 72 hours of incubation. The deer velvet extract promoted embryonic development and maintained blastocyst development rates similar to the control when embryos were challenged with hydrogen peroxide. Gene expression of anti-oxidative enzymes in blastocysts was not significantly different between deer velvet treatment groups. The deer velvet extract thus relieved oxidative stress on embryos and supported blastocyst development in vitro.
This document discusses various biomolecules associated with pregnancy, focusing on Pregnancy Associated Glycoproteins (PAGs). It describes how PAGs are secreted by trophoblast cells of the placenta and can be detected in maternal blood and milk as early as 15-30 days into pregnancy, making them useful for early pregnancy diagnosis. The document outlines the 22 known bovine PAGs and how PAG-1 in particular has properties that make it a reliable biomarker for developing commercial diagnostic kits to detect pregnancy more accurately and earlier than traditional methods.
This document provides an overview of milk kefir, including its microbial composition, biological activities, and related products. It discusses that kefir is produced by fermenting milk with kefir grains containing a symbiotic culture of lactic acid bacteria, acetic acid bacteria, and yeasts. The microbial composition of kefir grains can vary depending on origin but typically includes Lactobacillus, Lactococcus, yeasts such as Kluyveromyces and Candida species. Kefir is considered probiotic due to its microorganisms and metabolites, and kefiran, a polysaccharide in kefir grains, possesses various biological activities. Kefir can be marketed as a natural probiotic beverage
Gonadal histo morphology and antifertility effects of bonny light crude oil i...Alexander Decker
This document summarizes a study that investigated the effects of ingesting Bonny Light Crude Oil (BLCO) on male fertility in rats. Rats were given varying doses of BLCO for 60 days, then sperm counts were analyzed and testes were examined histologically. Sperm counts significantly decreased in treated rats compared to controls. Histological examination found degenerative and necrotic changes in testes of treated rats, including fewer sperm-producing cells and damage to Sertoli cells that support sperm maturation. The results suggest that ingesting BLCO has adverse effects on the male reproductive system and fertility in rats.
This study examined 1213 human embryos at the cleavage and blastocyst stages to assess the relationship between embryo morphology and chromosomal abnormalities. At the cleavage stage, 84% of embryos were found to be chromosomally abnormal, even among those with the best morphological grades. However, at the blastocyst stage, chromosomal abnormalities were significantly less common in embryos with optimal morphology and more common in poor-quality embryos. Nonetheless, many chromosomally abnormal blastocysts still received the highest morphological scores, particularly for trisomy 21 which is sometimes compatible with live birth. In summary, chromosomal abnormalities had little effect on cleavage stage morphology but began to impact blastocyst appearance, though the effects were often subtle
- Animal development is defined as the progressive changes in size, shape, and function that occur during an organism's life through which its genetic potential is translated into mature systems.
- Maternal genes play a major role in early development by providing nutritive and determinative materials to the egg during oogenesis, organizing the egg for subsequent development. Mutations in these maternal effect genes can impact offspring viability without affecting the mother.
- After fertilization, zygotic gene expression begins from the embryo genome, directing cell differentiation and organ formation under genetic control. Homeotic genes determine body segmentation by transforming one segment into another.
presented by HAFIZ M WASEEM
university of education LAHORE Pakistan
i am from mailsi vehari and studied in lahore
bsc in science college multan
msc from lahore
This study evaluated and optimized somatic embryogenesis procedures for commercial cacao clones. In the first part, eight clones and a control clone were assessed using a modified standard protocol to produce primary and secondary somatic embryos. The second part aimed to improve primary somatic embryogenesis efficiency for two important clones, CCN 51 and CCN 10, by testing different explant types and culture conditions. The results showed that it is possible to produce plants through somatic embryogenesis, though efficiency varies greatly between genotypes requiring protocol optimization. Secondary somatic embryogenesis can increase embryo production ten-fold. The importance of adapting protocols to each genotype is discussed.
Phenotypic Plasticity, CYP19A1 Pleiotropy, and Maladaptive Selection in Devel...J Patrick Malone
The contribution of evolutionary psychology to the study of development and psychopathology depends on adherence to the principles of evolutionary biology. The human brain evolved because selection favored neither size nor complexity but
instead the phenotypic plasticity supporting cognitive flexibility. Cell proliferation, migration, elongation, synaptogenesis, synaptic pruning, apoptosis, and myelination occur at varying rates during asynchronous phases of development throughout
the brain. Developmentally sensitive periods result from phenotypic plasticity and are vital for adaptation to the environment. The biological systems surrounding the CYP19A1 gene provide mechanisms for neuroprotection and targeted neuronal
debridement in response to environmental stress, uniting selection with developmental biology. Updates to Dunbar’s original hypothesis with current primatological data, inclusion of total brain mass, and the introduction of CYP19A1 orthology from
nine primate species yields a linear regression, R2 = .994, adjusted R2 = .989, F(3, 5) = 143.758, p < .001.
Meat and milk from farmed animals including livestock (cattle, goat and buffalo) and poultry are sources of high quality protein and essential amino acids, minerals, fats and fatty acids, readily available vitamins, small quantities of carbohydrates and other bioactive components.1 The Food and Agriculture Organization (FAO) 2008 estimate shows that meat consumption has grown with increase in population. The average global per capita meat consumption is 42.1 kg/year with 82.9 kg/year in developed and 31.1 kg/year in developing countries in a recommended daily animal-sourced protein per capita of 50 kg per year2. Milk on the other hand is consumed in various forms: liquid, cheese, powder, and cream at a global per capita consumption of 108 kg per person per year which is way below the FAO recommended daily consumption of 200 kg.
Meat and milk from farmed animals including livestock (cattle, goat and buffalo) and poultry are sources of high quality protein and essential amino acids, minerals, fats and fatty acids, readily available vitamins, small quantities of carbohydrates and other bioactive components.1 The Food and Agriculture Organization (FAO) 2008 estimate shows that meat consumption has grown with increase in population. The average global per capita meat consumption is 42.1 kg/year with 82.9 kg/year in developed and 31.1 kg/year in developing countries in a recommended daily animal-sourced protein per capita of 50 kg per year2. Milk on the other hand is consumed in various forms: liquid, cheese, powder, and cream at a global per capita consumption of 108 kg per person per year which is way below the FAO recommended daily consumption of 200 kg.
Este documento trata sobre tiristores. Explica que los tiristores son dispositivos de potencia populares en electrónica de potencia que permiten alcanzar altas potencias. Describe los diferentes tipos de tiristores como SCR, GTO, TRIAC y DIAC. También incluye información sobre su funcionamiento, parámetros y aplicaciones prácticas como control de iluminación.
Preguntas y respuestas sobre la ley de ilicitos cambiariosOscar Barreto
La Ley de Ilícitos Cambiarios castiga la compra, venta y transferencia de divisas diferentes al bolívar que no se realicen a través de organismos autorizados como Cadivi. Las sanciones incluyen multas equivalentes al doble del monto de la operación ilegal y penas de prisión de 2 a 7 años dependiendo de la gravedad del delito. Quedan prohibidas las operaciones en efectivo, cheques u otros instrumentos en moneda extranjera que superen los 10.000 dólares anuales sin declaración.
El documento presenta un diagnóstico de la situación socioeducativa de la Escuela Básica "Blanca Guevara de Balan" ubicada en Barrancas del Orinoco, Municipio Sotillo, Estado Monagas. Se analizan aspectos del contexto familiar, escolar y comunitario, destacando la diversidad cultural y las necesidades de fortalecer las estrategias del lenguaje para mejorar la lectura y escritura en los estudiantes del tercer grado. El diagnóstico concluye que es necesario implementar acciones pedagógicas que motiven
The document describes the Atlas Elektronik multipurpose heavy weight torpedo. It can be launched from submarines, ships, or shore and target underwater or surface targets. It has various homing and guidance modes including active/passive acoustic homing and intercept/wake homing. It has a high speed, long range, and standard 533mm calibre. Atlas Elektronik provides full life support and the torpedo is integrated on various submarine classes. It has nearly 50 years of experience with over 10,000 test firings.
Este documento es un formulario para presentar reclamos y solicitudes de servicio relacionados con transacciones nacionales con tarjetas de crédito, débito y cuentas financieras ante el Banco de Venezuela. El formulario solicita información del cliente, detalles del reclamo o servicio solicitado, y una declaración jurada del cliente. El cliente debe adjuntar la documentación requerida y explicar los motivos de la solicitud.
Este convenio cambiario establece que las personas jurídicas y naturales pueden mantener fondos en moneda extranjera en cuentas bancarias en Venezuela procedentes de proyectos de inversión pública, títulos de deuda emitidos por el gobierno o actividades de exportación. Los bancos deben reportar mensualmente al Banco Central de Venezuela sobre estos fondos en moneda extranjera y las empresas estatales pueden destinar hasta el 5% de sus saldos mensuales en divisas a la compra de títulos de deuda emitidos
Este documento es un formulario para presentar reclamos y solicitudes de servicio relacionados con transacciones nacionales de tarjetas de crédito, débito y cuentas financieras ante el Banco de Venezuela. El formulario solicita información del cliente, detalles del reclamo o servicio solicitado, y una declaración jurada del cliente. El cliente debe adjuntar documentación que respalde su solicitud y firmar el formulario.
The M777 lightweight 155mm field howitzer is a mobile artillery piece that provides reliable fire support. It is lightweight and portable, allowing it to be deployed quickly by land, sea, or air to various terrains. Through the use of titanium and aluminum alloys, the M777 is highly mobile while maintaining accuracy. It has been battle-tested in Afghanistan, firing over 40,000 rounds in harsh desert conditions.
Este documento trata sobre unas jornadas de charla sobre el gas natural y su incidencia en la sociedad. Se presenta la introducción sobre las características básicas del gas natural, sus principales reservas y usos industriales y domésticos. Luego, se plantea el problema de que en la unidad educativa Mariaguzman de Marcano no hay distribución directa de gas para el comedor, lo que causa falta de rendimiento y dificultades en el transporte de cilindros que no satisfacen la demanda. Esto genera pérdidas
Este documento trata sobre unas jornadas de charla sobre el gas natural y su incidencia en la sociedad. Presenta una introducción sobre las características, principales reservas y usos del gas natural. También plantea un problema en la unidad educativa MARIAGUSMAN DE MARCANO donde no cuentan con distribución directa de gas para el comedor, lo que causa falta de rendimiento y dificultades en el transporte de cilindros. Por lo tanto, se debe implementar líneas de gas directo para resolver este problema que afecta a los estudiantes
Este documento proporciona instrucciones para operar una transmisión Eaton Fuller Roadranger de 10 velocidades. Explica las posiciones de la palanca de cambios, los controles de cambio, el procedimiento de doble embrague, y consejos para la operación segura de la transmisión. También incluye información sobre el modelo de transmisión, características, lubricación y mantenimiento preventivo.
Copia de sordos con discapacidad angelica rodriguezOscar Barreto
El documento describe las deficiencias auditivas y los trastornos asociados, incluyendo el compromiso auditivo, las dificultades en el aprendizaje, el déficit de atención e hiperactividad, el compromiso cognitivo y el autismo. Para cada trastorno, se describen las características clave y las estrategias y necesidades educativas especiales recomendadas.
Comision estadal de deficiencias auditivas cedaOscar Barreto
La Comisión Estatal de Deficiencias Auditivas (CEDA) tiene como misión definir un plan conjunto con otras organizaciones para la prevención, detección, evaluación diagnóstica y atención educativa integral de personas con deficiencias auditivas. Su visión es fortalecer el subsistema de educación especial en el área de deficiencias auditivas para garantizar la integración social, laboral y de acuerdo con las necesidades de las personas. Los objetivos de CEDA incluyen fortalecer el subsistema de educación especial y el área de deficiencias audit
Lengua de señas venezolana e interpretaciónOscar Barreto
Este documento resume las diferencias entre el lenguaje oral y la lengua de señas venezolana. Explica que la lengua de señas se codifica de manera espacio-gestual y se decodifica visualmente, mientras que el lenguaje oral se codifica y decodifica de forma auditiva. También describe los niveles lingüísticos de ambos sistemas y explica por qué la lengua de señas es importante para las personas sordas como medio para comunicarse y desarrollar su identidad cultural.
Abordaje pegagogico del niño sordociegoiOscar Barreto
El C.A.I.D.V.A. es el Centro de Atención Integral de Deficiencias Visuales, una unidad del Ministerio de Educación de Venezuela que brinda atención educativa integral a personas ciegas y con deficiencias visuales a través de dos líneas: atención directa en el centro para desarrollar habilidades, y como unidad de apoyo trabajando con familias y otros centros para la atención temprana de niños de 0 a 6 años.
Este documento describe los procedimientos y mediciones esenciales para la reconstrucción de motores, incluyendo mediciones del bloque, pistones, cigüeñal, culata y sus componentes. Explica cómo medir el ovalamiento, conicidad, diámetro y planitud utilizando herramientas como micrómetros, comparadores y calibradores. También cubre la medición de holguras, juegos y ajustes para garantizar un ensamblaje correcto y un funcionamiento óptimo del motor reconstruido.
Este documento presenta los estatutos de una Organización Comunitaria de Vivienda (OCV) en Venezuela. Describe la estructura organizativa de la OCV, incluyendo la Asamblea General, la Asamblea de Delegados, la Junta Directiva y varios comités. También especifica los requisitos y deberes de los asociados, el proceso de toma de decisiones en las asambleas, y las atribuciones de la Asamblea General. El objetivo principal de la OCV es mejorar las viviendas de los asociados y promover la acción
GraphSummit Singapore | The Future of Agility: Supercharging Digital Transfor...Neo4j
Leonard Jayamohan, Partner & Generative AI Lead, Deloitte
This keynote will reveal how Deloitte leverages Neo4j’s graph power for groundbreaking digital twin solutions, achieving a staggering 100x performance boost. Discover the essential role knowledge graphs play in successful generative AI implementations. Plus, get an exclusive look at an innovative Neo4j + Generative AI solution Deloitte is developing in-house.
Observability Concepts EVERY Developer Should Know -- DeveloperWeek Europe.pdfPaige Cruz
Monitoring and observability aren’t traditionally found in software curriculums and many of us cobble this knowledge together from whatever vendor or ecosystem we were first introduced to and whatever is a part of your current company’s observability stack.
While the dev and ops silo continues to crumble….many organizations still relegate monitoring & observability as the purview of ops, infra and SRE teams. This is a mistake - achieving a highly observable system requires collaboration up and down the stack.
I, a former op, would like to extend an invitation to all application developers to join the observability party will share these foundational concepts to build on:
Building RAG with self-deployed Milvus vector database and Snowpark Container...Zilliz
This talk will give hands-on advice on building RAG applications with an open-source Milvus database deployed as a docker container. We will also introduce the integration of Milvus with Snowpark Container Services.
“An Outlook of the Ongoing and Future Relationship between Blockchain Technologies and Process-aware Information Systems.” Invited talk at the joint workshop on Blockchain for Information Systems (BC4IS) and Blockchain for Trusted Data Sharing (B4TDS), co-located with with the 36th International Conference on Advanced Information Systems Engineering (CAiSE), 3 June 2024, Limassol, Cyprus.
A tale of scale & speed: How the US Navy is enabling software delivery from l...sonjaschweigert1
Rapid and secure feature delivery is a goal across every application team and every branch of the DoD. The Navy’s DevSecOps platform, Party Barge, has achieved:
- Reduction in onboarding time from 5 weeks to 1 day
- Improved developer experience and productivity through actionable findings and reduction of false positives
- Maintenance of superior security standards and inherent policy enforcement with Authorization to Operate (ATO)
Development teams can ship efficiently and ensure applications are cyber ready for Navy Authorizing Officials (AOs). In this webinar, Sigma Defense and Anchore will give attendees a look behind the scenes and demo secure pipeline automation and security artifacts that speed up application ATO and time to production.
We will cover:
- How to remove silos in DevSecOps
- How to build efficient development pipeline roles and component templates
- How to deliver security artifacts that matter for ATO’s (SBOMs, vulnerability reports, and policy evidence)
- How to streamline operations with automated policy checks on container images
Let's Integrate MuleSoft RPA, COMPOSER, APM with AWS IDP along with Slackshyamraj55
Discover the seamless integration of RPA (Robotic Process Automation), COMPOSER, and APM with AWS IDP enhanced with Slack notifications. Explore how these technologies converge to streamline workflows, optimize performance, and ensure secure access, all while leveraging the power of AWS IDP and real-time communication via Slack notifications.
Threats to mobile devices are more prevalent and increasing in scope and complexity. Users of mobile devices desire to take full advantage of the features
available on those devices, but many of the features provide convenience and capability but sacrifice security. This best practices guide outlines steps the users can take to better protect personal devices and information.
Maruthi Prithivirajan, Head of ASEAN & IN Solution Architecture, Neo4j
Get an inside look at the latest Neo4j innovations that enable relationship-driven intelligence at scale. Learn more about the newest cloud integrations and product enhancements that make Neo4j an essential choice for developers building apps with interconnected data and generative AI.
Pushing the limits of ePRTC: 100ns holdover for 100 daysAdtran
At WSTS 2024, Alon Stern explored the topic of parametric holdover and explained how recent research findings can be implemented in real-world PNT networks to achieve 100 nanoseconds of accuracy for up to 100 days.
Unlocking Productivity: Leveraging the Potential of Copilot in Microsoft 365, a presentation by Christoforos Vlachos, Senior Solutions Manager – Modern Workplace, Uni Systems
Dr. Sean Tan, Head of Data Science, Changi Airport Group
Discover how Changi Airport Group (CAG) leverages graph technologies and generative AI to revolutionize their search capabilities. This session delves into the unique search needs of CAG’s diverse passengers and customers, showcasing how graph data structures enhance the accuracy and relevance of AI-generated search results, mitigating the risk of “hallucinations” and improving the overall customer journey.
Goodbye Windows 11: Make Way for Nitrux Linux 3.5.0!SOFTTECHHUB
As the digital landscape continually evolves, operating systems play a critical role in shaping user experiences and productivity. The launch of Nitrux Linux 3.5.0 marks a significant milestone, offering a robust alternative to traditional systems such as Windows 11. This article delves into the essence of Nitrux Linux 3.5.0, exploring its unique features, advantages, and how it stands as a compelling choice for both casual users and tech enthusiasts.
Large Language Model (LLM) and it’s Geospatial Applications
Duszewskaetal212
1. Journal of Animal and Feed Sciences, 21, 2012, 217–233
Obtaining farm animal embryos in vitro*
A.M. Duszewska1,3
, Ł. Rąpała1
, P. Trzeciak1
, S. Dąbrowski1
and A. Piliszek2
1
Warsaw University of Life Sciences, Division of Histology and Embryology,
Department of Morphological Sciences, Faculty of Veterinary Medicine
Nowoursynowska 159, 02-776 Warsaw
2
Institute of Genetics and Animal Breeding, Polish Academy of Sciences,
Experimental Embryology Department
Jastrzębiec, Postępu 1, 05-552 Magdalenka, Poland
(Received 22 March 2012; revised version 21 May 2012; accepted 18 June 2012)
ABSTRACT
This paper reviews the basic knowledge about obtaining farm animal embryos in vitro
with special focus on differences among species and application of this procedure in the future.
In vitro production of farm animal embryos consists of in vitro maturation (IVM) of oocytes, in
vitro fertilization (IVF) of matured oocytes, and in vitro culture (IVC) of embryos. Oocytes can
be collected from live animals (by laparotomy, laparoscopy, Ovum Pick Up) or from slaughtered
ones (by puncture, sectioning). Usually immature oocytes are isolated, and during IVM they reach
maturity. Matured oocytes are cultured with sperm (IVF), leading to the formation of zygotes. In
the case of fertilization problems (horse, pig), intracytoplasmic sperm injection is used. The zygotes
are usually cultured (IVC) to the morula and blastocyst stages. These embryos can be transferred to
recipients or frozen/vitrified. Offspring have been obtained after transfer of cattle, sheep, goat, pig
and horse embryos. This procedure can be used in animal breeding, biotechnology, medicine, and
basic research.
KEY WORDS: oocytes, sperm, embryos, in vitro, farm animals
INTRODUCTION
Obtaining farm animal embryos in vitro is a biotechnique used both
commercially (in vitro production, IVP), in basic research, as well as in infertility
*
Support by COST DPN/DWM/MZ/5670/08/09
3
Corresponding author: e-mail: duszewskaanna@hotmail.com
2. 218 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
treatment (Gordon, 2004; Thibier, 2005; Betteridge, 2006; Galli and Lazzari, 2008;
Duszewska et al., 2010; Figueirêdo Freitas et al., 2010; Dang-Nguyen et al., 2011;
Zhao et al., 2011). Commercially, it is used in animal breeding and biotechnology,
hence it is called in vitro production of embryos (IETS-www.iets.org). The goal
of this procedure is entirely different in basic research and infertility treatment,
where it plays the same role as the assisted reproduction techniques (ART) used
in humans (Gardner et al., 2011). For this reason it is hard to call this biotechnique
in vitro production of embryos, and instead it is called obtaining embryos in vitro
(Duszewska et al., 2010).
Obtaining farm animal embryos in vitro is a multi-step procedure comprising:
in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC)
(Scheme 1). Offspring of some farm animal species have been obtained after
transfer of embryos obtained in vitro to recipients, including sheep (Gandolfi et
al., 1987), cattle (Lu et al., 1988), pigs (Mattioli et al., 1989), and goats (Crozet et
al., 1993). For many years, obtaining horse embryos in vitro presented the biggest
challenge. In 1991 two foals were born, albeit from naturally matured oocytes that
were fertilized and cultured in vitro (Palmer et al., 1991).
The most important factors responsible for success in obtaining embryos in
vitro are: the source of oocytes and sperm (including the age and physiological
state of female and male), technical factors, and highly skilled laboratory and
veterinary staff. The key technical factors are: media for maturation, fertilization,
and embryo culture (containing water, amino acids, vitamins, hormones,
growth factors, and additives obtained from albumin, serum, etc.), conditions of
maturation, fertilization, and culture (temperature, CO2
level, humidity), and the
timing of oocyte maturation, fertilization, and embryo development (Duran, 2000;
Camargo et al., 2006).
All of these requirements are characteristic of the respective farm animal
species and have been described in detail in the literature: cattle (Hoshi, 2003;
Lonergan, 2007; Lonergan and Fair, 2008), pigs (Abeydeera, 2002; Nagai et al.,
2006; Lonergan and Fair, 2008; Gil et al., 2010; Dang-Nguyen et al., 2011), sheep
and goats (Cognie et al., 2003; Paramio, 2010), and horses (Squires at el., 2003;
Betteridge, 2006, 2007; Hinrichs, 2010).
PROCEDURE OF OBTAINING EMBRYOS IN VITRO FROM FARM ANIMALS
In vitro maturation of oocytes (IVM)
Oocytes are collected from the follicles located in the cortex of the ovary.
There are differences in the localization of the cortex among farm animal species.
In cattle, sheep, goats, and pigs the cortex is the outermost layer, however, in
3. 219DUSZEWSKA A.M. ET AL.
Scheme1.Obtainingoffarmanimalsembryosinvitro.Inthefirststage(IVM),oocytesarecollectedfromthedonors(liveorslaughtered)and
subsequentlyaresubjecttoinvitromaturation.Inthenextstageinvitrofertilizationmaturedoocytesareincubatedwithspermwhichleadsto
fertilization.Inthelaststageinvitroculture,theembryosobtainedfromIVFareculturedinvitro,usuallyuptoblastocyststage,whentheycanbe
transferredtorecipients(ET)orcanbefrozenorvitrifiedandusedatalaterdate
4. 220 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
mares the cortical layer is located inside the ovary, which poses some problems in
oocyte collection.
Oocytes can be collected from live or slaughtered animals. The methods used
for collection from live animals are: laparotomy (in all species), laparoscopy
(sheep, goats and swine, as well as horses and cattle) and ovum pick up (OPU)
(cattle, horses). OPU entails puncture of ovarian follicles. In sheep and goats,
laparoscopy combined with OPU is a popular method (Pieterse et al., 1991; Carter
et al., 2000; Galli et al., 2001; Abeydeera, 2002; Baldassarre et al., 2002; Cogine
et al., 2003, 2004; Squires et al., 2003; Colleoni et al., 2007; Paramio, 2010).
Oocytes can be collected from females during natural hormonal cycles or after
hormonal stimulation. Pregnant or young females can be used as oocyte donors
(Romaguera, 2011). Mature oocytes (Metaphase II) can be isolated, although in the
majority of cases, immature ones are harvested (Miyano, 2003). The frequency of
oocyte isolation is often a function of ovarian follicle development and maturation,
which is species specific (Hendriksen, 2000; Hirao, 2011; Sirard, 2011).
The most frequently used methods of obtaining material from slaughterhouse
samples are: puncture of an ovarian follicle with a needle connected to a suction
pump (in all species), sectioning of ovaries (mostly in mares), and isolation of
ovarian follicles and subsequent collection of oocytes (in all species) (Abeydeera,
2002; Cogine et al., 2003; Squires et al., 2003).
Immature oocytes are surrounded by several tightly packed layers of granulosa
cells called the corona radiata, with which they form the cumulus-oocyte complex
(COC), and a cluster of cells forming the cumulus oophorus (Abeydeera, 2002;
Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006; Lonergan, 2007; Lonergan
and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-Nguyen et al., 2011). In
mares, oocytes can be surrounded by both tightly packed and dispersed layers of
the corona radiata, but both types of oocytes are competed for maturation (Squires
et al., 2003; Betteridge, 2006; Hinrichs, 2010, 2010a).
Immature oocytes are at the prophase stage of the first meiotic division, namely
in GV (germinal vesicle) stage. Oocyte maturation encompasses simultaneous
nuclear and cytoplasmic changes. The key factors regulating meiosis are MPF
(maturation promoting factor) and MAP(mitogen activated protein kinase), whose
activity is necessary for the oocytes to reach the metaphase II stage (Gilchrist and
Thompson, 2007).
In the majority of mammals, resumption of meiosis in vitro is spontaneous
and is commenced by germinal vesicle break down (GVBD) and completed by
reaching the metaphase II stage of meiotic division by the oocyte, the stage when
it can be properly fertilized (Miyano and Manabe, 2007). Cytoplasmic maturation
includes a number of modifications - both morphological (organelle migration,
increase in the number of certain organelles, such as mitochondria, and formation
5. 221DUSZEWSKA A.M. ET AL.
of the cytoskeleton and spindle apparatus) and biochemical (cell cycle regulation,
metabolic processes, accumulation of informational and storage material) (Watson,
2007).
The granulosa cells of the corona radiata play an important role in the
maturation of the oocyte, being responsible for contact of the oocyte with the
environment and, therefore, mediating the hormonal and metabolic regulation of
this process (Abeydeera, 2002; Cognie et al., 2003; Hoshi, 2003; Nagai et al.,
2006; Lonergan, 2007; Lonergan and Fair, 2008; Gil et al., 2010; Paramio, 2010;
Dang-Nguyen et al., 2011).
The matured oocyte is characterized by an evenly dispersed corona radiata and
cytoplasm without any signs of degeneration, fragmentation, or vacuolization. In
some farm animal species, the first polar body can be observed, however, in the
majority, it is usually obscured by the opaque oocyte cytoplasm (due to its high
lipid content), therefore, the stage of maturation is determined indirectly on the
basis of dispersion of the granulosa cells of the corona radiata (Abeydeera, 2002;
Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006; Lonergan, 2007; Lonergan
and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-Nguyen et al., 2011).
There are also differences in the timing of oocyte maturation in vitro among
species (Banwell and Thompson, 2008). In cows, sheep, and goats, oocytes reach
maturity (metaphase II), after 24 h culture, and in horse, between 24 and 30 h.
Maturation takes the longest in pig oocytes, where M II is reached within 40-48 h
of culture (Abeydeera, 2002; Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006;
Lonergan, 2007; Lonergan and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-
Nguyen et al., 2011).
In vitro fertilization of matured oocytes
The next step in obtaining embryos in vitro is the fertilization of matured
oocytes. Usually, thawed sperm is used for fertilization, however, since in sheep
and horses problems with freezing sperm occur, fresh sperm is used in these
species (Morrell and Rodriguez-Martinez, 2010) Also, XY sperm separation can
be performed and the obtained sperm used for fertilization (Seidel, 2003; Cran,
2007).
Spermatozoa used in in vitro fertilization have to undergo capacitation, which
starts with removal of the glycoprotein coat. This subsequently leads to structural
modifications and changes in the intramembrane protein topography of the head
and tail of the spermatozoon. The changes in the sperm head enable the acrosomal
reaction to occur, whereas changes in the tail lead to hyperactivation (augmented
movement of the sperm). In vitro capacitation is a reversible process, in contrast
to the acrosomal reaction, which is an irreversible process allowing the sperm to
6. 222 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
penetrate the zona pellucida (Tulsiani et al., 2007; Bailey, 2010; Gadella, 2011).
The acrosomal reaction consists of the release of enzymes that allow the sperm
to penetrate the zona pellucida. The entry of sperm leads to oocyte activation,
completion of the II meiotic division resulting in the extrusion of the II polar body.
Subsequently, two pronuclei are formed, DNA replication taking place in each
of them, and the first mitotic division is initiated when the chromosomes of the
oocyte and sperm form a common metaphase plate. Differences among species
can pertain to asynchrony in pronuclei formation and the moment of replication
initiation (Gadella, 2010; Ikawa et al., 2010; Visconti and Florman, 2010; Tulsiani
et al., 2011).
Fertilization in pigs and horses poses many problems, because polyspermy (the
entry of more than one sperm into the oocyte) can often be observed both in vivo
and in vitro. Interestingly, in the case of in vitro matured oocytes, the frequency
of polyspermy is much lower than in in vivo matured oocytes. Polyspermy leads
to creation of polyploid embryos. In some cases triploid embryos can develop
up to the blastocyst stage, however, at later stages they degenerate and undergo
resorption. Because of the high lipid content in the cytoplasm it is not possible to
evaluate the quality of these embryos in the way it is done with human embryos
(Abeydeera, 2002; Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006; Lonergan,
2007; Lonergan and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-Nguyen et
al., 2011).
In pigs, polyspermy has been successfully overcome by modification of the
fertilizationmedium,andinhorsesitiscircumventedbytheuseofintracytoplasmic
sperm injection (ICSI) (Squires, 1996). In pigs, chromatin decondensation defects
are sometimes observed in the formation of the male pronucleus (Gil et al.,
2010).
In horses, in vitro fertilization problems are related to sperm entry into the
oocyte, and, specifically, to the penetration of the zona pellucida (Alm et al., 2001;
Roasa et al., 2007). To date there are only two foals that were obtained with the
use of conventional fertilization (Palmer, 1991). To circumvent this problem, the
ICSI technique, that is, the introduction of the spermatozoon into the cytoplasm
of in vitro matured oocyte, is routinely used (Palermo et al., 1992; Squires et al.,
2003), and has been further modified by the use of piezoICSI (Choi et al., 2002;
Galli et al., 2002).
The ICSI technique can be used in all farm animal species (Garcia-Rosello et
al., 2009), although in cattle, sheep, goats, and pig in vitro fertilization poses no
problems (Goto and Yanagita, 1995; Catt et al., 1996; Kolbe and Holtz, 2000).
In all farm animal species, the timing of fertilization is similar and takes place
18-20 h from insemination. Slight differences are the result of introduction of the
sperm to the oocyte in the ICSI procedure, which can lead to earlier formation of
7. 223DUSZEWSKA A.M. ET AL.
the zygote (Goto and Yanagita, 1995; Catt et al., 1996; Kolbe and Holtz, 2000;
Garcia-Rosello et al., 2009; Nakai et al., 2011).
In vitro culture of embryos
The last stage of obtaining embryos in vitro is their culture, that is the
development of the embryos from zygote to the morula and blastocyst stages. This
encompasses nuclear and cytoplasmic modifications and their mutual interactions,
changes in embryo metabolism, embryonic genome activation, modifications
of gene expression, formation of the morula and compaction of blastomeres,
formation of the blastocyst with a characteristic structure named the blastocyst
cavity that arises as a result of cavitation, and formation of additional structures
such as the capsule in the horse embryos (Abeydeera, 2002; Cognie et al., 2003;
Hoshi, 2003; Nagai et al., 2006; Lonergan, 2007; Lonergan and Fair, 2008; Gil et
al., 2010; Paramio, 2010; Dang-Nguyen et al., 2011). The capsule is a structure
located between trophoblast and zona pellucida, which is generated by the embryo.
Its role is to protect the embryo and it is found in embryos obtained both in vitro
and in vivo (Tremoleda et al., 2003).
One of the most important processes that take place during embryonic
development both in vitro and in vivo is the acquisition of control by the
embryonic genome - the maternal-zygotic transition (MET) (Vigneault et al.,
2004). Activation of the zygotic genome takes place at the 4-blastomere stage in
pigs, and the 8-16 blastomere stage in cattle, sheep, goats, and horses. Up until the
moment of embryonic genome activation, embryonic development is based on the
material stored during oocyte maturation (Sirard, 2010).
Differences occur among species in in vitro obtained blastocyst morphology
that concern the size of the embryos and the number of blastomeres forming the
inner cell mass (ICM) and the trophectoderm. The significant differences in the
number of blastomeres in horse embryos between blastocysts obtained in vivo
(1761 blastomeres) and in vitro (255 blastomeres) deserve special attention. In
other species the observed differences are not that great (Pomar et al., 2005).
The duration of embryonic development up to the blastocyst stage is
characteristic of a given species and is connected with the length and number
of consecutive cell cycles. Pig embryos have the shortest development time,
reaching blastocyst stage in only 144 h, while cattle, ovine, and goat blastocysts
are formed within 168 h. Equine embryos develop the longest (168-192 h of
culture) (Abeydeera, 2002; Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006;
Lonergan, 2007; Lonergan and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-
Nguyen et al., 2011).
8. 224 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
Embryo transfer
After the transfer of farm species embryos obtained in vitro to recipients,
the percentage of offspring is lower than in case of embryos obtained in vivo
(Abeydeera, 2002; Cognie et al., 2003; Hoshi, 2003; Nagai et al., 2006; Lonergan,
2007; Lonergan and Fair, 2008; Gil et al., 2010; Paramio, 2010; Dang-Nguyen et
al., 2011).
The percentage of calving, kidding, or foaling after transfer of in vivo and in
vitro embryos is, respectively, cattle, 70 vs 45; sheep, 75 vs 30; horses, 75 vs 20;
goats, 75 vs 30; pigs, 60 vs 5. In the case of pigs the percentage is extremely low
(Pomar et al., 2005).
Embryos obtained in vitro can be frozen or vitrified and later transferred to
recipients. Their developmental abilities after transfer are much lower, however,
than non-frozen embryos obtained in vitro. In the majority of farm animal species,
progeny is obtained after transfer of embryos frozen at the early or mature
blastocyst stage. On the other hand, much less offspring can be obtained after
transfer of frozen, morula-stage in vitro embryos. Great hopes lie in vitrification,
which allows for higher survival rates of embryos and, moreover, enables obtaining
more progeny after their transfer (Dobrinsky, 2002; Vajta and Kuwayama, 2006).
CRYOPRESERVATION OF EMBRYOS AND OOCYTES
Cryopreservation of oocytes and embryos is performed in two ways: slow
freezing and vitrification. Slow freezing is a conventional method based on a low
concentration of cryoprotectant and slow temperature reduction. Vitrification is
an alternative method characterized by a high cryoprotectant concentration and
ultra-rapid cooling rates, which makes it possible to avoid water precipitation,
thus preventing intracellular ice crystal formation. In this case, the short time
of vitrification and elimination of costly programmable freezing equipment is
of great importance. There are many modifications of freezing and vitrification
procedures (Vajta and Kuwayama, 2006; Ledda et al., 2007; Pereira and Marques,
2008; Prentiz and Anzar, 2011; Saragusty and Arav, 2011).
Interspecific differences exist in cryopreservation resistance of both oocytes
and embryos. Oocytes can be cryopreserved at the GV stage and in metaphase
II, but compared with embryos, oocytes are much more sensitive. As a result of
cryoconservation they develop morphological and functional defects (Martins
et al., 2005; Contreras et al., 2008; Pereira and Marques, 2008), therefore, the
cryopreservationefficiencyislow.Thespecificityofoocytecryopreservationisdue
to the granulosa cells surrounding them; because of their role in oocyte maturation
they can not be removed. Further problems with oocyte cryopreservation are
9. 225DUSZEWSKA A.M. ET AL.
related to the morphology of the metaphase II oocyte cell membrane and cortical
granules and spindle apparatus (Pereira and Marques, 2008).
With the use of the conventional, slow method of freezing oocytes, offspring
have been obtained from many species, such as cattle (Otoi et al., 1996), sheep
(Woods et al., 2004), horse (Maclellan et al., 2002). After vitrification, offspring
have been obtained in cattle (Sripunya et al., 2010), pigs (Liu, et al., 2008), sheep
(Succu et al., 2007), and horse (Bogliolo et al., 2006).
Cryopreservation of farm animal embryos obtained in vitro does not pose as
many problems as that of oocytes, however, even in that case the efficiency of
freezing depends on the species, developmental stage of the embryos, and their
origin (in vitro or in vivo), as well as the method of cryoconservation and choice
of cryoprotectant. Cattle and sheep embryos are the most resistant, the least - pig
and horse embryos (Pereira and Marques, 2008). In the 1980s and 1990s, progeny
was obtained after freezing and thawing of farm animal embryos. Also after
vitrification of in vitro embryos, progeny of many farm animals has been obtained
(Dobrinsky, 2002; Vajta and Kuwayama, 2006; Saragusty and Arav, 2011).
DEVELOPMENT POTENTIAL OF EMBRYOS OBTAINED IN VITRO
The aim of the procedure of in vitro embryo production is to obtain a large
number and high quality of blastocysts. The number of embryos obtained depends
on the number of collected oocytes and their quality. On the other hand, the
quality of the blastocysts also depends on the conditions during oocyte maturation
and embryo culture and on the bull’s influence. Both quality and quantity of
blastocysts determines the number of offspring and its healthiness after transfer of
those embryos into recipients (van Wagtendonk-de Leeuw et al., 2000; Rizos et
al., 2002; Lonergan et al., 2003; McEvoy et al., 2006; Hansen et al., 2010; Lazzari
et al., 2010; Romaguera et al., 2010).
The development potential of embryos obtained in vitro is lower in comparison
with embryos obtained in vivo due to suboptimal conditions during oocyte
maturation, their subsequent fertilization, and embryonic development (Wright
and Ellington, 1995; Young et al., 1998; Lonergan et al., 2006; Metwally and
Ledger, 2011).
Evaluation of embryos is based mainly on the timing of mitotic cleavage
and on embryo morphology. In the morphological aspect, during the oocyte and
early embryonic development stages, one can observe inhibition of development,
fragmentation, vacuolization, and lysis. Analysis of embryonic development
indicates that in all species of farm animals, high mortality occurs at the early
stages of in vitro development. Among the types of death of single blastomeres,
and sometimes of all the blastomeres of the embryo, the most prominent are
10. 226 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
necrosis and senescence. Far less frequent is programmed cell death (apoptosis),
which can be observed mainly in horse embryos. More recently other types of cell
death have been attracting greater attention, specifically, autophagy and mitotic
catastrophe (Okada et al., 2004). One can not exclude other, so far unknown, types
of cell death (Betts et al., 2001; Okada et al., 2004).
These events do not always lead to embryo death, sometimes they will only
cause perturbations of development, which can become manifest much later, after
implantationorduringpostnataldevelopment.Amongthereasonsofhighmortality
and occurrence of disorders are: changes of metabolism during oocyte maturation
and embryo development, anomalous activation of the embryonic genome and
defective gene expression, chromosomal aberrations (polyploidy, aneuploidy,
mixploidy) (van Wagtendonk-de Leeuw et al., 2000). Epigenetic modifications, i.e.
theposttranslationalchangesingeneexpression(includinggenomicimprinting)that
can occur in vitro present an interesting problem. Epigenetic modifications during
maturation and embryonic development, and probably also during fertilization,
can alter preimplantation, postimplantation, and also postnatal development of
individuals. This can lead to various disorders, including: hydrocephalus, brain
hypoplasia or damage, abnormal limb development, heart and liver enlargement,
respiratory problems, as well as large offspring syndrome (LOS). Epigenetic
modifications can lead to the death of individuals in the pre- and postnatal period
(resorption, abortion, or death shortly after birth) (Young et al., 2000; Weaver et
al., 2009; Bartolomei and Ferguson-Smith, 2011; Li and Sasaki, 2011).
It should be stressed, however, that in most cases the progeny obtained after
transfer of the farm animal embryos obtained in vitro is indistinguishable both
anatomically and physiologically from that obtained under natural conditions.
APPLICATION OF PROCEDURES FOR OBTAINING EMBRYOS IN VITRO
FROM FARM ANIMALS
Obtaining livestock embryos in vitro has applications in animal breeding,
biotechnology, medicine, and basic research (Betteridge, 2006; Hansen, 2006;
Galli and Lazzari, 2008; Duszewska et al., 2010).
In animal breeding the procedure of obtaining embryos in vitro can be used
as an alternative technique to insemination or a multiple ovulation and embryo
transfer (MOET) programme, which consists of inducing multiple ovulations and
transferring embryos to recipients. It is possible to use females only as oocyte
donors without the burden of the further stages of this procedure. Immature or
pregnant females can also be used as oocyte donors. Oocytes and embryos can be
frozen or vitrified, which enables banking them (Gardner et al., 2011).
11. 227DUSZEWSKA A.M. ET AL.
Obtaining embryos in vitro is of the greatest importance in biotechnology
because this technique facilitates embryonic and somatic cloning, creating
transgenic individuals and their clones, and also creating chimeras. The great
hopes of human medicine lie in use of pigs as organ donors in xenotransplantation
(Duszewska et al., 2010).
Basic research in both oocyte maturation, oocyte fertilization and early
embryonic development is an important field that makes use of embryos obtained
in vitro. Early development of bovine embryos is similar to human embryos and,
therefore, cattle is considered to be a model species (Niemann et al., 2000).
In recent years more attention has been paid to the application of this
biotechnique in infertility treatment in farm animals, where it plays the same
role as assisted reproduction techniques (ART) used in humans. There are many
indications connected with farm animal infertility for the application of the in
vitro embryo obtaining procedure, including ICSI. Moreover, in special cases, the
microsurgical epididymal sperm aspiration (MESA) or testicular sperm aspiration
(TESA) (testicle biopsy) procedures can be applied, with subsequent use of ICSI
(Greve and Callesen, 2005; Duszewska et al., 2010).
Recommendations for use of procedures for obtaining embryos in vitro can
include:
1. Oviductal obstruction; in this case females are used only as oocyte donors.
2. The use of females only at the most desired moment, which is facilitated by
cryoconservation of gametes and embryos, that is, freezing and vitrification
of immature and mature oocytes and embryos, which enables banking them
similarly to freezing sperm, and for subsequent transfer of embryos to the
mother or surrogate mother.
3. Sex-linked genetic aberrations; in this case it is possible to regulate the sex in
a herd by use of sexed semen for in vitro fertilization.
4. Carrier-state of lethal genes can be detected at the early stages of embryonic
development and makes it possible to eliminate those embryos from the
population.
5. Polyspermy, that is, the penetration of more than one sparmatozoa into
the oocyte, generates polyploidy, e.g., triploid embryos, which develop
only until the blastocyst stage and undergo resorption at the later stages of
development.
6. Immunological infertility.
7. Insufficient number of motile spermatozoa (oligospermia) or excessive
thickness of the zona pellucida, which prevents sperm penetration, are
indications for ICSI.
12. 228 FARM ANIMALS EMBRYOS OBTAINING IN VITRO
8. Preimplantation genetic diagnosis (PGD) can supplement prenatal diagnostics,
which is a separate field in human medicine in infertility treatment. Among
the many directions of PGD, the most important include diagnostics of
chromosomal aberrations, including hereditary diseases.
9. Others.
Thanks to progress in many fields of science it has become possible to
treat infertility not only in humans, but also in farm animals by use of assisted
reproduction techniques and techniques used in biotechnology.
CONCLUSIONS
The procedure of obtaining farm animal embryos in vitro is commonly used
and is considered to be one of the most important biotechniques in reproduction
since it combines animal breeding with biotechnology as well as human and
veterinary medicine. Obtaining embryos in vitro is of the greatest importance in
biotechnology because this technique facilitates embryonic and somatic cloning,
creating transgenic individuals, their cloning, and also creating chimeras. The great
hopes of human medicine lie in use of pigs as organ donors in xenotransplantation.
Obtaining farm animal embryos in vitro may be used in infertility treatment.
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