ABSTRACT
Objective: This paper establishes levels and patterns of ability and willingness to pay (AWTP) for contraceptives, and associated factors. Study design: A three-stage cluster and stratified sampling was applied in selection of enumeration areas, households and individuals in a baseline survey for a 5-year Family planning programme. Multivariable linear and modified Poisson regressions are used to establish factors associated with AWTP. Results: Ability to pay was higher among men (84%) than women (52%). A high proportion of women (96%) and men (82%) were able to pay at least Ug Shs 1000 ($0.27) for FP services while 93% of women and 83% of men who had never used FP services will in future be able to pay for FP services costed at least Shs 2000 ($0.55). The factors independently associated with AWTP were lower age group (<25 years), residence in urban areas, attainment of higher education level, and higher wealth quintiles. Conclusion: AWTP for FP services varied by different measures. Setting the cost of FP services at Shs 1000 ($0.27) will attract almost all women (96%) and most of men (82%). Key determinants of low AWTP include residence in poor regions, being from rural areas and lack of/low education. Implications statement: Private providers should institute price discrimination for FP services by region, gender and socio-economic levels. More economic empowerment for disadvantaged populations is needed if the country is to realise higher contraceptive uptake. More support for total market approach for FP services needed
Subject(s)
Aptitude , Cleavage Stage, Ovum , Contraceptive Agents , Ambulatory Care Facilities , Uganda , Women , MenABSTRACT
No presente estudo, utilizou-se a melatonina e a proteína específica do oviduto (pOSP) nos meios de maturação in vitro. Foram avaliadas a expansão do complexo cumulus-ovócito (CCOs), as concentrações intracelulares de espécies reativas de oxigênio (ROS) e o desenvolvimento embrionário nos diferentes grupos (C = controle; T1 = somente com melatonina; T2 = com melatonina e pOSP e T3 somente com pOSP). No tocante à expansão do CCOs, houve diferença (P<0,05) dos valores obtidos no grupo C em relação aos valores médios dos grupos T1, T2 e T3, porém não houve diferença entre os valores obtidos nos tratamentos (P>0,05). Na dosagem de ROS, não houve diferença entre os valores médios obtidos no grupo C (26,4±10,9) e o valor verificado no grupo T1 (23,4±7,8), porém no grupo T2 (21,3±9,7) o valor médio mostrou-se satisfatório em relação ao valor do grupo C. No entanto, o valor médio do grupo T3 (16,6±10,5) foi o que demonstrou resultado mais satisfatório quando comparado aos demais grupos (P<0,05). A produção de embriões foi avaliada por meio da taxa de clivagem. Não houve diferença (P >0,05) entre os valores obtidos entre o grupo C (48,9 %) e os valores verificados nos grupos T1 (51,5 %), T2 (50 %), T3 (57,7 %), nem destes entre si. Este estudo permitiu concluir que a proteína específica do oviduto recombinante e a melatonina foram eficientes em melhorar a expansão dos CCOs. Além disso, as células tratadas com pOSP mostraram-se com menor quantidade de ROS, podendo a pOSP ser considerada um antioxidante proteico.(AU)
The present study used melatonin and recombinant oviduct specific protein (pOSP) in in vitro maturation medium (IVM). The expansion of the cumulus-oocyte complexes (COCs), the intracellular concentrations of reactive oxygen species (ROS) and embryo development of the different groups were evaluated (C = control; T1 = melatonin; T2 = melatonin and pOSP and T3 = pOSP). Regarding the COCs expansion, the groups T1, T2 and T3 showed satisfactory results compared with group C (P<0.05), but there was no difference between treatments (P>0.05). In the ROS dosage, there was no difference between the mean values obtained in group C (26.4 ± 10.9) and group 1 (23.4 ± 7.8). However, in group 2 (21.3 ± 9.7), the average value was found to be satisfactory in relation group C. Despite that, the average value of treatment 3 (16.6 ± 10.5) was the most satisfactory result found compared to the other groups (P<0.05). The production of embryos was evaluated by cleavage rate, there was no difference between the values obtained in group C and the values recorded in groups T1 (51.5 %), T2 (50 %), T3 (57.7 %), and among them. This study showed that the pOSP and the melatonin were effective in the improvement of the expansion of COCs cells. In addition, the cells that were treated with pOSP presented a lower amount of ROS, allowing the pOSP to be considered a proteic antioxidant.(AU)
Subject(s)
Animals , Female , Embryonic Development , Fallopian Tubes/chemistry , Melatonin/administration & dosage , Swine , Antioxidants , Cleavage Stage, Ovum , Embryo Culture Techniques/veterinary , In Vitro Techniques/veterinaryABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of the rate of morphologically normal sperm (MNS) on the clinical outcomes of conventional in vitro fertilization (IVF) in patients with one retrieved oocyte.</p><p><b>METHODS</b>From January 2013 to January 2015, a total of 256 couples with one retrieved oocyte underwent conventional IVF in our center. According to the rate of MNS, the patients were divided into two groups: MNS < 4% (134 cycles) and MNS ≥ 4% (122 cycles). We compared the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo between the two groups. A total of 75 fresh embryo transfer cycles were performed, 43 in the MNS < 4% group and the other 32 in the MNS ≥ 4% group. We also compared the rates of implantation, clinical pregnancy and abortion between the two groups.</p><p><b>RESULTS</b>There were no statistically significant differences between the two groups in the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo (P > 0.05). The rates of implantation, clinical pregnancy and abortion exhibited no remarkable differences either in the fresh embryo transfer cycles between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The rate of MNS does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte.</p>
Subject(s)
Female , Humans , Male , Pregnancy , Abortion, Spontaneous , Cleavage Stage, Ovum , Embryo Implantation , Fertilization , Fertilization in Vitro , Methods , Oocyte Retrieval , Pregnancy Rate , Single Embryo Transfer , Sperm Count , Spermatozoa , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To explore the developmental potential of embryos at different developmental days and provide evidence for blastocyst culture of non-top quality cleavage stage embryos in frozen-thawed embryo transfer (FET) cycles.</p><p><b>METHODS</b>The clinical data of 687 FET cycles were retrospectively analyzed. According to the embryo freezing time, the patients were divided into day 5 (D5) blastocyst group (n=87), day 6 (D6) blastocyst group (n=111) and day 3 cleavage-stage embryo (D3) group (n=489) with hormone replacement cycles or natural cycles for endometrial preparation. The clinical pregnancy rates, miscarriage rates, and implantation rates were compared between the 3 groups.</p><p><b>RESULTS</b>The clinical pregnancy rate, miscarriage rate and implantation rate per transfer were 58.6%, 9.8%, and 42.9% in D5 group, 32.4%, 19.4%, and 23.3% in D6 group, and 44.9%, 16.4%, and 26.9% in D3 group, respectively. The clinical pregnancy rate and implantation rate were significantly higher in D5 group than in the other two groups (P<0.05).</p><p><b>CONCLUSION</b>The D5 blastocysts derived from non-top quality D3 embryos after cryopreservation can have better clinical outcomes than those derived from D3 cleavage-stage embryos and D6 blastocysts, and are therefore a better option than D3 cleavage-stage embryos in FET cycles.</p>
Subject(s)
Female , Humans , Pregnancy , Abortion, Spontaneous , Blastocyst , Cleavage Stage, Ovum , Cryopreservation , Embryo Implantation , Embryo Transfer , Pregnancy Rate , Retrospective StudiesABSTRACT
Background: Clarithromycin is a macrolide antibiotic used to treat common infections including respiratory tract, skin and Helicobacter pylori. This work investigates whether the administration of clarithromycin to pregnant females during the cleavage phase of gestation was associated with a risk of miscarriages and offspring morphological malformation and skeletal anomalies, histological changes and DNA fragmentation of embryos and liver of pregnant rats. Two major groups of pregnant albino rats were used. The animals of the control group received distilled water from the 1[st] to 7[th] days of gestation.One subgroup [C1] was sacrificed on the 8[th] day; and the other [C2] was sacrificed on the 20[th] day of gestation. The treated group was drenched 45 mg/kg clarithromycin [therapeutic dose] suspension from 1[st] to 7[th] day of gestation. The first subgroup [T1] was sacrificed on the 8th day and the other [T2] was sacrificed on the 20th day of gestation
Results: The obtained results showed a decrease in maternal body weight gain, increase in the rate of abortion, resorption and growth retardation of fetuses and some malformation in the skeletal system of the treated group. Histopathological studies of pregnant and fetal rats revealed congestion and dilatation of the central vein, fatty degeneration of the hepatocytes and severe DNA fragmentation
Subject(s)
Animals, Laboratory , Pregnancy, Animal , Rats , Cleavage Stage, Ovum/drug effects , DNA Fragmentation , Liver , Musculoskeletal SystemABSTRACT
From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. [...] During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.
Do Trópico de Capricórnio ao Equador, sabe-se que a sazonalidade no gato domestico é ausente, i.e., estes animais são considerados reprodutores não sazonais nestas regiões. [...] O objetivo deste experimento foi determinar a porcentagem de clivagem e formação de mórulas e blastocistos produzidos a partir de oócitos recuperados de ovários de gatas em três condições - folicular, lútea ou inativa - durante duas estações reprodutivas pelas quais gatas passam na região sudeste do Brasil (22°53'09" S e 48°26'42" O) - estação não reprodutiva (ENR), que compreende os meses de janeiro a março; e estação reprodutiva (ER), agosto à outubro. Trinta gatas foram castradas. [...] Durante a ENR, de um total de 272 (inativo), 162 (lútea) e 134 (folicular) oócitos fertilizados, a porcentagem de clivagem de zigotos, formação de mórulas e de blastocistos derivados de ovários inativos foi 24,63, 16,54 e 8,09 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 21,6, 12,96 e 8,64, e para aqueles provenientes de ovários na fase folicular, foi de 24,62, 16,41 e 8,21. Considerando a ER, de um total de 102 (inativo), 198 (lútea) e 86 (folicular) oócitos fertilizados, a frequência relativa (%) de zigotos clivados, mórulas e blastocistos derivados de ovários na condição inativa foi de 64,7, 41,17 e 23,53 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 64,14, 40,41 e 23,73, e para aqueles provenientes de ovários na fase folicular, foi de 63,95, 39,54 e 24,41. Os resultados deste experimento demonstraram que não houve diferença estatística significante (P < 0.05) na frequência de embriões clivados e na formação de mórulas e blastocistos quando comparadas as três condições ovarianas dentro da mesma estação. Entretanto, a ER apresentou resultados melhores considerando as taxas de clivagem e formação de mórula e de blastocisto se comparada à ENR.
Subject(s)
Animals , Cats , Blastocyst , Cleavage Stage, Ovum , Follicular Phase , Cats/embryology , Luteal Phase , Morula , In Vitro Oocyte Maturation Techniques/veterinary , Breeding , Oocyte Retrieval/veterinary , Reproduction/physiologyABSTRACT
Introdução: a biópsia embrionária tem como objetivo selecionar embriões geneticamente normais. Essa seleção ocorre por meio de testes genéticos pré-implantacionais. Espera-se, com isso, uma diminuição dos riscos de doenças genéticas e um aumento das taxas de implantação em fertilização in vitro. Objetivo: verificar, por meio de revisão bibliográfica, qual técnica de biópsia embrionária é considerada mais apropriada para feitura de testes genéticos pré-implantacionais. Método: pesquisa bibliográfica, na forma de revisão de publicações científicas, por meio das redes US National Library of Medicine (Pubmed), Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Google Acadêmico e Biblioteca Virtual em Saúde (BVS). Resultados e conclusão: existem três maneiras de efetuar a biópsia para reprodução humana assistida. A primeira consiste em retirar o primeiro e/ou o segundo corpúsculo polar estruído pelo oócito. Também se pode fazer a biópsia a partir de um blastômero do embrião em estágio de clivagem ou usar cinco a dez células do trofoectoderma de blastocisto. Normalmente as técnicas usadas para o diagnóstico são PCR, Fish, CGH array e SNP array, entre outras. Acredita-se que a biópsia de blastocistos é a melhor técnica para manter o potencial de implantação embrionária. Essa tendência se justifica por causa da maior quantidade de material genético disponível em fase avançada de desenvolvimento embrionário. Admite-se que nessa fase a incidência de mosaicismo seja menor em relação à biópsia de blastômeros, com consequente aumento na eficácia dos testes genéticos. Outra questão importante é que na biópsia de blastocistos as células são retiradas do trofoectoderma, enquanto que na biópsia em estágio de clivagem a remoção de um blastômero pode prejudicar o desenvolvimento embrionário.
Introduction: the embryo biopsy aims to select genetically normal embryos. This selection occurs through pre- implantation genetic testing. It is expected the reduction of risk ofgenetic disorders and increase implantation rates in IVF.Objective: to verify, through bibliographical revision, which embryo biopsy technique is considered more suitable for pre-implantation genetic diagnosis. Method: bibliographical research, in the form of literary review of scientific publications via networks, US National Library of Medicine (Pubmed), Latin-American Literature and Caribbean Health Sciences (Lilacs), Google Scholar and Virtual Health Library. Results and conclusion: there are three ways to perform the biopsy on assisted human reproduction.The first one consists in removing the 1st and/or 2nd polar body (if there wasfertilization). You can also perform the biopsy from the one blastomere of embryo cleavage stage or use 5-10 trophoectoderm cells blastocyst. Usually the techniques used for diagnosticpurpose are PCR, Fish, CGH array, SNP array and others. Nowadays it is believed that blastocyst biopsy is the best technique in order to maintain the embryonic implantation. This tendency is justified by the larger amount of genetic material available in an advancedstage of embryonic development. It is assumed that in this stage the incidence of mosaicism is reduced with the consequent increase in the effectiveness of genetic testing. Another important question is that the blastocyst biopsy cells are removed from the trophoectoderm while inbiopsy incleavage stage, the removal of one blastomere can impair embryonicdevelopment.
Subject(s)
Humans , Biopsy/methods , Choice Behavior , Embryo, Mammalian/cytology , Genetic Testing/methods , Blastocyst/cytology , Blastomeres/cytology , Cleavage Stage, Ovum , Embryo, Mammalian/pathology , Embryo Implantation/physiologyABSTRACT
Melatonin is a scavenger agent that has been used to promote in vitro embryo development. This study was designed to show the effects of melatonin on the quality and quantity rate of preimplantation mouse embryo development and pregnancy. In this experimental study, super ovulated, mated mice were killed by cervical dislocation to collect two-cell zygotes from the oviduct of pregnant 1 day NMRI mice. Zygotes were cultured to the hatching blastocyst stage and the numbers of embryos at different stages were recorded under an inverted microscope. The cleavage rates of two-cell zygotes were assayed until the blastocyst and hatching blastocyst stage in drops of T6 medium that contained either melatonin [1, 10, and 100×10[6], 10 and 100×10[9] M] or no melatonin. The cell numbers of blastocysts were determined by differential staining, implantation outcomes were studied, and development and pregnancy rate were compared by the Chi-square [development] and Fisher's exact [pregnancy rate] tests. The addition of 10 and 100 nM melatonin to the embryo culture media promoted the development of the two-cell stage embryos to blastocyst and hatching blastocysts [p<0.01] and caused a significant increase in total cell number [TCN], trophoectoderm [TE], and inner cell mass [ICM] of the blastocysts [p<0.01]. A difference was observed in the percentage of transferred embryos that were successfully implanted between the control and treatment groups [p<0.05]. The data indicate that 10 and 100 nM of melatonin positively impact mouse embryo cleavage rates, blastocyst TCN, and their implantation. Therefore, melatonin at low concentrations promotes an embryonic culture system in mice
Subject(s)
Animals, Laboratory , Embryonic Development , Embryo Implantation , Cleavage Stage, Ovum , MiceABSTRACT
<p><b>OBJECTIVE</b>To analyze sex chromosome mosaicisms in early cleavage-stage human embryos and blastocysts with poor embryo quality score based on the numbers of pronucleus(PN) zygotes using X,Y dual color fluorescence in situ hybridization (FISH), and to discuss the possible mechanisms.</p><p><b>METHODS</b>Fresh or frozen-thawed early cleavage-stage human embryos and blastocysts with poor embryo quality score not suitable for embryo transfer were studied with dual color FISH.</p><p><b>RESULTS</b>Double signal rate of 2PN among early cleavage-stage embryos was 66.67%, which was significantly higher than 1PN and 3PN embryos. Single signal rate of 1PN early cleavage-stage embryos was 90.41%, which was significantly higher than 2PN and 3PN ones. Three signal rate of 3PN early cleavage-stage embryos was 28.00%, which was significantly higher than 1PN and 2PN ones. Double signal rate of 3PN ones was 46.00%, which was significantly higher than 1PN ones. The polyploid rate of frozen-thawed early cleavage-stage embryos was 23.53%, which was slightly higher than that of fresh embryos, but with no statistical significance. The mosaicism rate of 24 blastocysts was 100.00% and the double signal dominant (≥ 50%) rate was 62.50%, which was significantly higher than the rate of early cleavage-stage embryos.</p><p><b>CONCLUSION</b>Using 2PN as the criterion for embryo quality score cannot guarantee the selection of normal fertilized embryo for transplantation. Frozen-thawed embryos may harbor more polyploid cells. To avoid the selection of embryos with abnormal chromosomes, combinations of pre-implantation genetic diagnosis (PGD) and prenatal diagnosis are necessary. Meanwhile, blastocysts with poor quality scores may provide an important source for embryo stem cells.</p>
Subject(s)
Humans , Blastocyst , Metabolism , Cleavage Stage, Ovum , Metabolism , Fertilization in Vitro , In Situ Hybridization, Fluorescence , Mosaicism , Embryology , Sex ChromosomesABSTRACT
OBJECTIVE: To evaluate the impact of sperm defect severity and the type of azoospermia on the outcomes of intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: This study included 313 ICSI cycles that were divided into two major groups according to the source of spermatozoa used for ICSI: 1) Ejaculated (group 1; n = 220) and 2) Testicular/Epididymal (group 2; n = 93). Group 1 was subdivided into four subgroups according to the results of the semen analysis: 1) single defect (oligo-[O] or astheno-[A] or teratozoospermia-[T], n = 41), 2) double defect (a combination of two single defects, n = 45), 3) triple defect (OAT, n = 48), and 4) control (no sperm defects; n = 86). Group 2 was subdivided according to the type of azoospermia: 1) obstructive (OA: n = 39) and 2) non-obstructive (NOA: n = 54). Fertilization (2PN), cleavage, embryo quality, clinical pregnancy and miscarriage rates were statistically compared using one-way ANOVA and Chi-square analyses. RESULTS: Significantly lower fertilization rates were obtained when either ejaculated sperm with triple defect or testicular sperm from NOA patients (63.4 + 25.9 percent and 52.2 + 29.3 percent, respectively) were used for ICSI as compared to other groups (~73 percent; P < 0.05). Epididymal and testicular spermatozoa from OA patients fertilized as well as normal or mild/moderate deficient ejaculated sperm. Cleavage, embryo quality, pregnancy and miscarriage rates did not differ statistically between ejaculated and obstructive azoospermia groups. However, fertilization, cleavage and pregnancy rates were significantly lower for NOA patients. CONCLUSION: Lower fertilization rates are achieved when ICSI is performed with sperm from men with oligoasthenoteratozoospermic and non-obstructive azoospermic, and embryo development and pregnancy rates are significantly lower when testicular spermatozoa from NOA men are used.
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Fertilization , Sperm Injections, Intracytoplasmic , Sperm Retrieval , Spermatozoa/physiology , Analysis of Variance , Azoospermia , Chi-Square Distribution , Cleavage Stage, Ovum , Ejaculation , Oligospermia , Pregnancy Outcome , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To assess the effects of the nuclear status of day 2 preembryos on day 3 embryo quality and implantation potential and to weigh its clinical value in the human in-vitro fertilization-embryo transfer (IVF-ET) program.</p><p><b>METHODS</b>Embryos obtained from 409 fresh conventional IVF-ET/ICSI cycles from July to October 2006 were assessed retrospectively. Day 2 preembryos were classified according to the number of nuclei in each blastomere in 3 groups: grade A with only mononucleated blastomeres, grade B with one or more blastomeres containing no visible nucleus, and grade C with one or more multinucleated blastomeres. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3 as well as of the pregnancy outcome and implantation potential of those in whom cohorts of similar nuclear scoring embryos were transferred.</p><p><b>RESULTS</b>There were fewer arrested embryos and more excellent embryos on day 3 in grade A than in grade B and C (P < 0.01), and so were there in grade B than in grade C (P < 0.01). Among the 234 cycles in which all the transfer embryos were derived from a similar day 2 nuclear scoring, 51 cycles originated from grade A embryos (group A) and 183 from grade B (group B), with similar clinical pregnancy rates between the two groups, while the implantation rate was higher in group A than in B (P < 0.05).</p><p><b>CONCLUSION</b>Day 2 nuclear scoring can be used to predict the devel- opment and implantation potential of embryos. A combined evaluation of day 2 nuclear scoring and day 3 embryo morphology helps identify the most viable embryos and reduce the number of embryos for transfer.</p>
Subject(s)
Adult , Female , Humans , Male , Pregnancy , Blastomeres , Cell Nucleus , Physiology , Cleavage Stage, Ovum , Embryo Implantation , Physiology , Embryo Transfer , Fertilization in Vitro , Pregnancy Outcome , Sperm Injections, IntracytoplasmicABSTRACT
The best developmental stage and the best thawing protocol suitable for cryopreservation of the early embryos are not well documented. The present study aimed at evaluating the effect of the ultra rapid cryopreservation [vitrification] technique, followed by slow or fast thawing protocol, on the fertilized ova, 4-cell embryos and morula. The vitrification method included equilibrating the ova in the vitrification solution [EFS40; consisted of 40% ethylene glycol, 30% Ficoll, 0.5 M sucrose in D-PBS] for 2 minutes before immersion in liquid nitrogen. Slow and fast thawing were done and the cryoprotectants were withdrawn by a hyperosmolar sucrose solution, which was then gradually diluted and replaced by culture medium. The best results were obtained with vitrification of the 4-cell embryos both with slow and fast thawing, which gave survival rate of 86% and 94%, and in vitro development rate of 74% and 80%, respectively. Fast thawing showed better survival rates [80%, 94%, 77%] and better in vitro development rates [60%, 80%, 63%] than those of slow thawing, following vitrification of the fertilized ova, 4-cell embryos and morula, respectively. These criteria of vitrification/ thawing could be inferred to the human 4-cell embryos in the IVF protocol
Subject(s)
Humans , Embryo Culture Techniques , Embryonic Development , Embryo Research , Cleavage Stage, Ovum , Vitrification , Fertilization in Vitro , Reproductive Techniques, AssistedABSTRACT
Exposure of either gametes or embryos to conditions and/or factors that generate oxidative stress has been associated with impaired early embryogenesis. The effects of reactive oxygen species (ROS) on mouse preimplantation development, depending of the ROS-concentration and time of exposition, were studied. Two-cell embryos were incubated with 5, 10, 25 and 50 microM of hydrogen peroxide (H2O2) for 30 and 60 minutes of exposition and allowed to develop for 72 h to study the quality of Development. The incubation with 50 microM H2O2 for 30 or 60 minutes, strongly inhibited the 2-cell embryo development as compared to the control (p < 0.001). Twenty-five microM H2O2 produced inhibition of blastocyst formation (p < 0.001) and 10 microM H2O2 significantly decreased the percentages of expanded and hatchedblastocysts, which resulted morphologically altered (p < 0.05 and p < 0.01, respectively). The higher H2O2 concentrations were able to elicit necrotic morphology in the 2-cell arrested embryos, while 10 microM H2O2 induced moderate damage with the arrested embryos partially fragmented. In conclusion, important causes for defective preimplantation development and for early embryo losses may be due to oxidative stress because early mouse embryos exposed to ROS for short times arrested at the first cellular cycle (2-cell) and/or impaired embryo differentiation and morphogenesis, being these effects ROS-concentration-dependent.
Subject(s)
Animals , Male , Female , Mice , Blastocyst/cytology , Blastocyst , Blastocyst/metabolism , Reactive Oxygen Species/toxicity , Cleavage Stage, Ovum , Hydrogen Peroxide/administration & dosage , Hydrogen Peroxide/toxicity , Embryonic Development , Embryo Transfer , Oxidative StressABSTRACT
To study the effects of mouse cytomegalovirus (MCMV) on the in vitro maturation, fertilization, cleavage and blastula formation of mouse oocytes, the immature oocytes were infected in vitro by MCMVs of different dosages (100 TCID(50), 10 TCID(50) and 1 TCID(50)). The oocytes were then observed for in vitro maturation, fertilization, cleavage and blastula formation and the ultrastructural changes after the culture with the viruses. Our results showed that no significant differences were found in IVM, IVF, cleavage and blastula formation among the groups treated with of virus of various dosages. And ultrastructural abnormality was observed in the oocytes treated by 100 TCID(50) of viruses. It is concluded that MCMV did not have any conspicuous effects on IVM, IVF, cleavage and blastula formation of murine immature oocytes.
Subject(s)
Blastocyst , Cells, Cultured , Cleavage Stage, Ovum , Cytomegalovirus Infections , Fertilization , Muromegalovirus/pathogenicity , Oocytes/cytology , Oocytes/growth & development , Oocytes/virologyABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of Er'zhi Tiangui Granule (ETG) in improving the quality of oocyte.</p><p><b>METHODS</b>Ninety mice were randomly divided into 6 groups. The number of high-quality oocytes was comparatively observed in the 1st experimental group and the 1st control group; the embryonic cleavage rate was observed in the 2nd experimental group and the 2nd control group and the quantity of insulin-like growth factor-1R mRNA (IGF-1R mRNA) expression in ovarian granular cells was determined by using in situ hybridization in the 3rd experimental group and the 3rd control group.</p><p><b>RESULTS</b>The high-quality oocytes rate, the embryonic cleavage rate and the quantity of IGF-1R mRNA expression in the three paired groups was (78 +/- 8)% vs (71 +/- 5)%, (88 +/- 3)% vs (83 +/- 5)%, 0.4890 +/- 0.0454 vs 0.4439 +/- 0.0283, respectively. The difference between the experimental groups to the respective control groups was significant (all P < 0.05), and positive correlation was shown between the high-quality oocytes rate and the quantity of IGF-1R mRNA expression.</p><p><b>CONCLUSION</b>The mechanism of ETG in improving the quality of oocyte may be related with the elevation of IGF-1R mRNA level in ovarian granular cells.</p>
Subject(s)
Animals , Female , Male , Mice , Cleavage Stage, Ovum , Physiology , Drugs, Chinese Herbal , Pharmacology , Gene Expression , Oocytes , Physiology , Ovary , Metabolism , RNA, Messenger , Genetics , Random Allocation , Receptor, IGF Type 1 , GeneticsABSTRACT
The interaction between molecular biology and embryology made an extensive progress in the research on gametogenesis, fertilization and early embryogenesis in mice. In this article, molecules involving in meiotic maturation and apoptosis of oocytes, sperm-oocyte interactions and early cleavage of fertilized embryos in mice are described including our recent following experiments. 1] Phosphatidylinositol 3-kinase and Akt participate in the follicle stimulating hormone-induced meiotic maturation of mouse oocytes. 2] Mos plays a crucial role in normal spindle and chromosome morphology and the reactivation of maturation promoting factor after first meiosis. 3] Follicular atresia is caused by apoptosis and the apoptosis associated with internucleosomal DNA fragmentation is directly regulated by the Fas-Fas ligand system. 4] Integrin alpha 6 beta 1 is involved in sperm-egg binding leading to fusion via direct association of the integrin alpha 6 with sperm. 5] MAP kinase cascade is activated at the M-phase and some MAP kinases other than ERKs are activated during early cleavage of fertilized eggs
Subject(s)
Animals, Laboratory , Apoptosis , Oocytes , Cleavage Stage, Ovum , Mice , Membrane GlycoproteinsABSTRACT
The study was carried out to determine the effectiveness and safety of the infrared 1.48 microm laser in cleavage stage mouse embryo biopsy, compared to the conventional acid Tyrode's solution. One hundred and thirty cryopreserved cleavage stage mouse embryos were included in the study. Fifty embryos were biopsied by acid Tyrode's solution. Forty-seven embryos were biopsied by the infrared 1.48 microm laser. Thirty-three embryos were incubated without biopsy as the control group. Thirteen of 50 embryos in the acid Tyrode's group and 16 of 47 in the laser assisted group became cavitating morulae on day 4, meanwhile 23 of 33 in the control group reached this stage. The blastocyst formation of acid Tyrode's, laser assisted and control group were 94.0, 97.8 and 100.0 per cent, respectively. The hatching rate of acid Tyrode's solution, laser assisted and control group were 78.7, 84.7 and 63.6 per cent, respectively. No significant difference in blastocyst formation and hatching rate was found. The percentage of grade I blastocysts in control group (96.9%) was significantly higher than those in acid Tyrode's solution (68.0%) and the laser assisted group (76.0%). There was no significant difference in the percentage of grade 1 blastocysts between the acid Tyrode's solution and the laser assisted group. In conclusion, the infrared 1.48 microm wavelength laser may be an alternative to acid Tyrode's solution in embryo biopsy.