The purpose of our study was to compare the protein patterns originating from fluids of mature an... more The purpose of our study was to compare the protein patterns originating from fluids of mature and immature human follicles in order to gain further insight into their biochemical composition. A total of 10 patients were stimulated for in-vitro fertilization (FVF) using different stimulation protocols. Follicular fluids were aspirated transvaginally and analysed microscopically for the presence of oocytes. Follicular fluids were stored at-18°C. Samples of 500 |il were processed for two-dimensional gel electrophoresis. Up to 60 proteins in various groups could be detected. Seven protein spots were selected for chemical analysis by cutting them out of the gels and subjecting them to internal amino add sequencing procedures. Our results can be summarized as follows: (i) major differences were not detected between the protein patterns from the various mature follicles of a particular patient, nor were significant differences observed in the proteins derived from follicular fluids collected from the seven patients with mature follicles; (ii) considerable differences were observed in the protein patterns derived from fluids of immature compared with mature follicles. Fluid from the three patients with immature follicles contained many fewer proteins, some of which were expressed at low levels. We conclude that the observed variations in protein composition of follicles of different developmental age reflect their physiological condition and serve as biomedical markers for follicular maturity.
Objective: To analyze the concentration of growth factors secreted by autologous cumulus cell clu... more Objective: To analyze the concentration of growth factors secreted by autologous cumulus cell clusters during in vitro human embryo co-culture and investigate the impact of culture supplementation with autologous cumulus cell clusters on embryo development and pregnancy rate as compared to standard culture procedure. Design: Randomized prospective comparative study. Setting: Private IVF clinic. Patients: 90 IVF patient couples ranging at the age between 35 and 40 (mean age of 36.5) agreed to participate in this study. In this IVF group, 64 first trials, 22 s trials and 4 third trials were included. 48 cases have been terminated at day 3 of culture and 42 of them have been continued up to day 5. Of the 48 cases assigned for day-3 culture, 208 MII-oocytes were randomly assigned for co-culture and 197 MII-oocytes were assigned for standard culture. The selection was done by alternating the oocyte immediately after stripping to co-culture or control, starting always with co-culture. Interventions: All samples were analyzed with Milliplex factor-specific magnetic-bead panels with phycoerythrin fluorescent factor-specific antibodies using immunofluorescence cytometry technology by Lab Supplies Inc. laboratories in Athens, Greece. The MFI (Mean Fluorescence Intensity) values were calibrated and the readings were translated in pg/ml and calculated against the background values of culture medium alone. The statistical analysis of all data were processed with the Two-tailed T-test form. Main outcome measures: 36 IVF cases terminated at day 3 were randomly chosen. Supernatants from 17 cases were analyzed for seven different growth factors, namely, epidermal growth factor (EGF), fibroblast growth factor 1 and 2 (FGF-1 and FGF-2), vascular endothelial growth factor A, C and D (VEGF-A, VEGF-C and VEGF-D) and Leptin. Supernatants from 19 cases were analyzed for Insulin-like Growth Factor I and II (IGF-I and IGF-II). Results: VEGF-A and VEGF-C were found at statistically significant levels (p < 0.005) in culture supernatant samples where the autologous cumulus clusters were present but were not detectable in supernatants of embryos cultured alone. IGF-I was detected at considerable levels in supernatant samples where the autologous cumulus clusters were present. IGF-I was not detectable in supernatant samples of embryos cultured alone. IGF-II was detected in significant amounts (p < 0.05) in samples of all groups. The highest concentration level of IGF-II was detected in supernatant samples of embryos in standard culture. Conclusion(s): The investigated factors, among other substances, may be causally connected to the beneficial effect observed on embryo development. Our findings suggest that co-culture with autologous cumulus cell clusters improves the outcome of embryo culture in IVF programs.
gestational sacs with fetal heart beat by transvaginal ultrasound examination in week 7. RESULTS:... more gestational sacs with fetal heart beat by transvaginal ultrasound examination in week 7. RESULTS: The number of good quality embryo was significantly higher in the TL group compared to control group (4.63AE2.22 VS 3.65AE1.84, p¼0.0014); pregnancy rate and implantation rate showed a tendency to be higher in the TL group compared to control group.
Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell ... more Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell determination after transplantation to either the anterior or mid-ventral region of the early embryo. By means of similar transplantations, we have tested the ability of polar plasm of Drosophila immigrans to induce the formation of pole cells in a Drosophila melanogaster embryo. After the transplantation of polar plasm, "hybrid" pole cells were found in which both pole cell-specific organelles, the polar granules and nuclear body, were structurally similar to those characteristic of the transplanted cytoplasm. In order to determine whether these hybrid cells can function as germ cell precursors, these cells were transplanted to the posterior tip of genetically marked embryos. Approximately 5% of the flies obtained from embryos receiving potential pole cells produce offspring derived from the induced pole cells. This result demonstrates that polar plasm can function in interspe...
Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the c... more Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the cleavage stage. Methods: Human triploid embryos at the 6-8 cell stage were split into twin embryos (D and R) and cultured in vitro to the blastocyst stage. Twin embryos were processed for nested multiplex PCR using fluorescently labeled primers of six selected polymorphic STR markers within the human HLA locus. Results: A total of 11 pairs of twin embryos were analyzed with PCR technology. From five pairs of twin blastocysts, the fluorograms provided detectable STR profiles for identical peak position between the twin embryos. Conclusion: This is the first report on twinned human embryos to prove monozygocity at the DNA level. Embryo splitting exhibits novel potential for future applications in assisted reproductive medicine.
MRI and sonography. The FSH oestradiol inhibin B Lh levels were studied. After stimulation, oocyt... more MRI and sonography. The FSH oestradiol inhibin B Lh levels were studied. After stimulation, oocytes were retrieved and IVF was attempted. RESULTS: During the menopausal hormonal therapy, the FSH level varied from 50 UI/ml to 170 UI/ml. MRI vascularization analyse proved the presence of new vascularization originating from the sub cutaneous vessels. The new vascularization attained the graft from the external side of the cortex. The doppler resistance index of the grafting vessels fell when follicular growth was observed. MRI and sonography never displayed any ovarian activity on the remainder ovary. Five months after the graft, the FSH level fell to 18UI/ml. Following the first stimulation, the oestradiol level went up then down, and only one atretic oocyte was retrieved, eight other stimulations were performed and one spontaneous cycle was observed. A maximum of nine oocytes per cycle were obtained, with 910 pg/ml oestradiol level.A total of 23 oocytes were obtained which 3 atretics, 3 pellucid fractures,2 germinal vesicles, 4 in metaphase I, 7 in metaphase II. Three embryos were transferred after ICSI, one of these needed IVM. The follicle's maximum diameter was 12 mm. Hormonal variations were sub normal during the spontaneous cycles. CONCLUSION: According to literature, two human pregnancies were obtained after orthotopic cryopreserved ovarian grafting. Heterotopic ovarian grafting is an other way of using ovarian tissue, proved by animal experimentation. This case reports the second human embryo obtained by heterotopic grafting of ovarian cryopreserved tissue, and analyses the new vascularization and the hormonal variation of the graft.
ss as: Ill s.2011.03 Abstract Objective: To prove monozygocity of human twin blastocysts derived ... more ss as: Ill s.2011.03 Abstract Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the cleavage stage. Methods: Human triploid embryos at the 6–8 cell stage were split into twin embryos (D and R) and cultured in vitro to the blastocyst stage. Twin embryos were processed for nested multiplex PCR using fluorescently labeled primers of six selected polymorphic STR markers within the human HLA locus. Results: A total of 11 pairs of twin embryos were analyzed with PCR technology. From five pairs of twin blastocysts, the fluorograms provided detectable STR profiles for identical peak position between the twin embryos. Conclusion: This is the first report on twinned human embryos to prove monozygocity at the DNA level. Embryo splitting exhibits novel potential for future applications in assisted reproductivemedicine. 2011 Middle East Fertility Society. Production and hosting by Elsevier B.V. All rights reserved.
... Beispiel: der chinesische Schrift-steller Kuo Mo-jo vor und nach Mao. -- lEin hochinteressant... more ... Beispiel: der chinesische Schrift-steller Kuo Mo-jo vor und nach Mao. -- lEin hochinteressantes Buch ft~r jeden, der (noch) Freude am Sprechen und Schrei-ben hat. ... Man erhiilt Informatiouen fiber lEngliseh, Deutsch, Franz6siseh, Russisch und Chinesiseh als Sprache. ...
The purpose of our study was to compare the protein patterns originating from fluids of mature an... more The purpose of our study was to compare the protein patterns originating from fluids of mature and immature human follicles in order to gain further insight into their biochemical composition. A total of 10 patients were stimulated for in-vitro fertilization (FVF) using different stimulation protocols. Follicular fluids were aspirated transvaginally and analysed microscopically for the presence of oocytes. Follicular fluids were stored at-18°C. Samples of 500 |il were processed for two-dimensional gel electrophoresis. Up to 60 proteins in various groups could be detected. Seven protein spots were selected for chemical analysis by cutting them out of the gels and subjecting them to internal amino add sequencing procedures. Our results can be summarized as follows: (i) major differences were not detected between the protein patterns from the various mature follicles of a particular patient, nor were significant differences observed in the proteins derived from follicular fluids collected from the seven patients with mature follicles; (ii) considerable differences were observed in the protein patterns derived from fluids of immature compared with mature follicles. Fluid from the three patients with immature follicles contained many fewer proteins, some of which were expressed at low levels. We conclude that the observed variations in protein composition of follicles of different developmental age reflect their physiological condition and serve as biomedical markers for follicular maturity.
Objective: To analyze the concentration of growth factors secreted by autologous cumulus cell clu... more Objective: To analyze the concentration of growth factors secreted by autologous cumulus cell clusters during in vitro human embryo co-culture and investigate the impact of culture supplementation with autologous cumulus cell clusters on embryo development and pregnancy rate as compared to standard culture procedure. Design: Randomized prospective comparative study. Setting: Private IVF clinic. Patients: 90 IVF patient couples ranging at the age between 35 and 40 (mean age of 36.5) agreed to participate in this study. In this IVF group, 64 first trials, 22 s trials and 4 third trials were included. 48 cases have been terminated at day 3 of culture and 42 of them have been continued up to day 5. Of the 48 cases assigned for day-3 culture, 208 MII-oocytes were randomly assigned for co-culture and 197 MII-oocytes were assigned for standard culture. The selection was done by alternating the oocyte immediately after stripping to co-culture or control, starting always with co-culture. Interventions: All samples were analyzed with Milliplex factor-specific magnetic-bead panels with phycoerythrin fluorescent factor-specific antibodies using immunofluorescence cytometry technology by Lab Supplies Inc. laboratories in Athens, Greece. The MFI (Mean Fluorescence Intensity) values were calibrated and the readings were translated in pg/ml and calculated against the background values of culture medium alone. The statistical analysis of all data were processed with the Two-tailed T-test form. Main outcome measures: 36 IVF cases terminated at day 3 were randomly chosen. Supernatants from 17 cases were analyzed for seven different growth factors, namely, epidermal growth factor (EGF), fibroblast growth factor 1 and 2 (FGF-1 and FGF-2), vascular endothelial growth factor A, C and D (VEGF-A, VEGF-C and VEGF-D) and Leptin. Supernatants from 19 cases were analyzed for Insulin-like Growth Factor I and II (IGF-I and IGF-II). Results: VEGF-A and VEGF-C were found at statistically significant levels (p < 0.005) in culture supernatant samples where the autologous cumulus clusters were present but were not detectable in supernatants of embryos cultured alone. IGF-I was detected at considerable levels in supernatant samples where the autologous cumulus clusters were present. IGF-I was not detectable in supernatant samples of embryos cultured alone. IGF-II was detected in significant amounts (p < 0.05) in samples of all groups. The highest concentration level of IGF-II was detected in supernatant samples of embryos in standard culture. Conclusion(s): The investigated factors, among other substances, may be causally connected to the beneficial effect observed on embryo development. Our findings suggest that co-culture with autologous cumulus cell clusters improves the outcome of embryo culture in IVF programs.
gestational sacs with fetal heart beat by transvaginal ultrasound examination in week 7. RESULTS:... more gestational sacs with fetal heart beat by transvaginal ultrasound examination in week 7. RESULTS: The number of good quality embryo was significantly higher in the TL group compared to control group (4.63AE2.22 VS 3.65AE1.84, p¼0.0014); pregnancy rate and implantation rate showed a tendency to be higher in the TL group compared to control group.
Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell ... more Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell determination after transplantation to either the anterior or mid-ventral region of the early embryo. By means of similar transplantations, we have tested the ability of polar plasm of Drosophila immigrans to induce the formation of pole cells in a Drosophila melanogaster embryo. After the transplantation of polar plasm, "hybrid" pole cells were found in which both pole cell-specific organelles, the polar granules and nuclear body, were structurally similar to those characteristic of the transplanted cytoplasm. In order to determine whether these hybrid cells can function as germ cell precursors, these cells were transplanted to the posterior tip of genetically marked embryos. Approximately 5% of the flies obtained from embryos receiving potential pole cells produce offspring derived from the induced pole cells. This result demonstrates that polar plasm can function in interspe...
Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the c... more Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the cleavage stage. Methods: Human triploid embryos at the 6-8 cell stage were split into twin embryos (D and R) and cultured in vitro to the blastocyst stage. Twin embryos were processed for nested multiplex PCR using fluorescently labeled primers of six selected polymorphic STR markers within the human HLA locus. Results: A total of 11 pairs of twin embryos were analyzed with PCR technology. From five pairs of twin blastocysts, the fluorograms provided detectable STR profiles for identical peak position between the twin embryos. Conclusion: This is the first report on twinned human embryos to prove monozygocity at the DNA level. Embryo splitting exhibits novel potential for future applications in assisted reproductive medicine.
MRI and sonography. The FSH oestradiol inhibin B Lh levels were studied. After stimulation, oocyt... more MRI and sonography. The FSH oestradiol inhibin B Lh levels were studied. After stimulation, oocytes were retrieved and IVF was attempted. RESULTS: During the menopausal hormonal therapy, the FSH level varied from 50 UI/ml to 170 UI/ml. MRI vascularization analyse proved the presence of new vascularization originating from the sub cutaneous vessels. The new vascularization attained the graft from the external side of the cortex. The doppler resistance index of the grafting vessels fell when follicular growth was observed. MRI and sonography never displayed any ovarian activity on the remainder ovary. Five months after the graft, the FSH level fell to 18UI/ml. Following the first stimulation, the oestradiol level went up then down, and only one atretic oocyte was retrieved, eight other stimulations were performed and one spontaneous cycle was observed. A maximum of nine oocytes per cycle were obtained, with 910 pg/ml oestradiol level.A total of 23 oocytes were obtained which 3 atretics, 3 pellucid fractures,2 germinal vesicles, 4 in metaphase I, 7 in metaphase II. Three embryos were transferred after ICSI, one of these needed IVM. The follicle's maximum diameter was 12 mm. Hormonal variations were sub normal during the spontaneous cycles. CONCLUSION: According to literature, two human pregnancies were obtained after orthotopic cryopreserved ovarian grafting. Heterotopic ovarian grafting is an other way of using ovarian tissue, proved by animal experimentation. This case reports the second human embryo obtained by heterotopic grafting of ovarian cryopreserved tissue, and analyses the new vascularization and the hormonal variation of the graft.
ss as: Ill s.2011.03 Abstract Objective: To prove monozygocity of human twin blastocysts derived ... more ss as: Ill s.2011.03 Abstract Objective: To prove monozygocity of human twin blastocysts derived from embryo splitting at the cleavage stage. Methods: Human triploid embryos at the 6–8 cell stage were split into twin embryos (D and R) and cultured in vitro to the blastocyst stage. Twin embryos were processed for nested multiplex PCR using fluorescently labeled primers of six selected polymorphic STR markers within the human HLA locus. Results: A total of 11 pairs of twin embryos were analyzed with PCR technology. From five pairs of twin blastocysts, the fluorograms provided detectable STR profiles for identical peak position between the twin embryos. Conclusion: This is the first report on twinned human embryos to prove monozygocity at the DNA level. Embryo splitting exhibits novel potential for future applications in assisted reproductivemedicine. 2011 Middle East Fertility Society. Production and hosting by Elsevier B.V. All rights reserved.
... Beispiel: der chinesische Schrift-steller Kuo Mo-jo vor und nach Mao. -- lEin hochinteressant... more ... Beispiel: der chinesische Schrift-steller Kuo Mo-jo vor und nach Mao. -- lEin hochinteressantes Buch ft~r jeden, der (noch) Freude am Sprechen und Schrei-ben hat. ... Man erhiilt Informatiouen fiber lEngliseh, Deutsch, Franz6siseh, Russisch und Chinesiseh als Sprache. ...
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