Partial zona dissection (PZD) of human oocytes facilitates sperm penetration through mechanically... more Partial zona dissection (PZD) of human oocytes facilitates sperm penetration through mechanically made holes in the zona pellucida. Only 1 of 69 eggs was damaged when sucrose was used to shrink the ooplasm during micromanipulation. The fertilization rate of micromanipulated oocytes in 18 couples with male factor infertility was 68% (34/50), which compared favourably with insemination of non-micromanipulated controls (21/45, 47%). PZD was advantageous in oligozoospermic patients, but not in cases of asthenozoospermia, combined semen problems or immunological infertility. Three twin and two singleton pregnancies resulted following replacement of 23 micromanipulated and eight control embryos in 14 patients. No differences in embryo morphology and development rates were found between the micromanipulated and control groups. The incidence of polyspermy in couples with abnormal semen analyses was relatively low (less than 20%) possibly due to partial activation of the oocytes following exposure to sucrose. Polyspermy was high (57%) in normozoospermic patients with either immunological infertility (n = 3) or failure of fertilization in previous cycles (n = 4). In the three immunological patients, nine of 11 hyaluronidase and sucrose-exposed control embryos fertilized and six implanted, possibly indicating that cumulus and corona cells are contributing factors inhibiting fertilization in such cases.
Partial zona dissection (PZD), a method using mechanical force to open the human zona pellucida, ... more Partial zona dissection (PZD), a method using mechanical force to open the human zona pellucida, and zona drilling, which uses acidic Tyrode's (AT) medium, were compared in 1-day-old oocytes prior to reinsemination. The incidences of monospermy and polyspermy were 13/54 (24%) and 14/54 (26%) following PZD and 6/46 (13%) and 8/46 (17%) following the use of AT medium. This compared favorably with conventional reinsemination: 15/161 (9%) monospermy and 4/161 (3%) polyspermy. Three of the 27 PZD embryos became blastocysts, while none of the AT-exposed embryos developed satisfactorily. Eleven male-factor couples had some of their oocytes randomly treated with PZD prior to insemination; each of the patients had non-micromanipulated control oocytes. Monospermic fertilization and cleavage (23/34; 68%) doubled (P less than 0.05) when PZD was compared with the control oocytes (10/30; 33%). Replacing two PZD and a single control embryo in two patients resulted in twin pregnancies. A third twin pregnancy was established following replacement of only micromanipulated embryos.
... Jacques Cohen1*2, Carlene Eisner, Hilton Kort, Henry Malter2*3, Joe Massey, Mary Pat Mayer an... more ... Jacques Cohen1*2, Carlene Eisner, Hilton Kort, Henry Malter2*3, Joe Massey, Mary Pat Mayer and Klaus Wiemer ... In comparison, fertile couples with unrestricted intercourse are believed to conceive in 25% of menstrual cycles, when usually only one oocyte is released ...
Correction of polyspermy through pronucleus extraction in the absence of membrane relaxants was a... more Correction of polyspermy through pronucleus extraction in the absence of membrane relaxants was applied to 25 polyspermic human zygotes. Nine zygotes survived the procedure, and seven cleaved normally (two of which were fixed for chromosome analysis); two proceeded to compact and one cavitated. Eighteen of the polyspermic zygotes (14 with three pronuclei and 4 with four pronuclei) were obtained from zona pellucida-intact oocytes, and seven (1 five-pronucleate 3 four-pronucleate, and 3 three-pronucleate) from previously zona-drilled oocytes. Survival and cleavage occurred in all groups except in four- and five-pronucleate zona-drilled zygotes. Criteria used to identify male pronuclei were (1) pronucleus-associated sperm tails, (2) increased pronucleus size, and (3) greater distance (relative to female pronuclei) from the second polar body. Sperm tails were never seen and pronucleus size usually was identical. Therefore, the third criterion was used, although its reliability should be further evaluated. Until complete pronucleus removal techniques and reliable pronucleus selection criteria are perfected, embryo replacement after polyspermy correction could result in aneuploidy and molar pregnancy.
Zygotes from in vitro fertilization patients (n = 116) were randomly allocated to culture in eith... more Zygotes from in vitro fertilization patients (n = 116) were randomly allocated to culture in either conventional plastic petri dishes or coculture on a monolayer of fetal bovine uterine fibroblasts. Embryos (n = 288) remained 26 to 32 hours in these culture systems. Video tape recording for later morphological analysis (11 parameters) was performed on 117 conventionally cultured and 104 cocultured embryos, shortly before replacement, by an independent observer, unaware of the culture conditions for each embryo. A significantly greater number of cocultured embryos (52%) had "good" morphology (zero or only one abnormal characteristic) as compared with conventionally cultured embryos (30%). The most outstanding morphological characteristic of cocultured embryos was the expanded appearance of their blastomeres. The incidence of implantation per embryo increased from 13% to 19% when the coculture rather than conventional culture system was used, and the incidence of ongoing pregnancy per patient after coculture doubled to 35%.
To establish guidelines for application of partial zona dissection, subzonal sperm insertion, and... more To establish guidelines for application of partial zona dissection, subzonal sperm insertion, and regular in vitro fertilization (IVF) in severe male factor patients. Two studies were performed: partial zona dissection and IVF was applied in 57 couples during the first period, and subzonal sperm insertion was also applied in a second group of 47 couples. Procedures were performed in an academic research environment. Couples who failed fertilization previously, others not acceptable for IVF, and a third group in whom IVF was expected to fail. Oocytes were micromanipulated with either partial zona dissection or subzonal sperm insertion, or the zona pellucida was left intact. Embryos were replaced in patients prophylactically treated with methylprednisolone and antibiotics. Because several microsurgical fertilization techniques are now available, this study was performed to compare sperm parameters, embryo morphology, fertilization, and implantation rates after application of two successful micromanipulation procedures. Twenty-one pregnancies were established in 104 patients, 5 definitely from subzonal sperm insertion and 4 from partial zona dissection. Patients who failed IVF before had a similar chance of pregnancy after the use of micromanipulation, as first time patients (9/53 versus 12/51). In a subgroup of 15 patients who failed IVF with insufficient numbers of motile sperm, fertilization was significantly higher after subzonal sperm insertion. Partially zona-dissected embryos from couples with severe teratozoospermia (less than or equal to 5% normal forms; strict criteria) had significantly more morphological abnormalities than those from patients with moderate teratozoospermia (6% to 10% normal forms). In severely teratozoospermic patients, significantly fewer partially zona-dissected than subzonally inserted embryos implanted. The decision of which micromanipulation method to perform can possibly be based on careful analysis of sperm morphology.
Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embry... more Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embryos with Vero cells in both Dulbecco's modified Eagle's medium/Ham's F12 (DMEM/F12) and human tubal fluid (HTF) culture medium. Two-cell CB6F1 mouse embryos were cultured either in the presence of Vero cells (group A) or in culture medium alone (group B). In DMEM/F12 significantly more morulae developed in group A than in group B on day 3 (91 versus 23%; P less than 0.01). In contrast, the mouse embryos grew rapidly in HTF and significant differences were noted only in later embryonic stages (on day 5; 86% and 50%, P less than 0.01 of group A and B respectively, hatching or hatched). Similar experiments using DF1 and ICR mouse strains also revealed enhanced embryo development in the presence of Vero cells. To determine whether the embryo-enhancing effects of Vero cells were due to the removal of toxins or to the secretion of embryotrophic factors, ICR mouse embryos were cultured in fresh media with cells (group A), without cells (group B) and in cell-free conditions using cell-conditioned media which were obtained in the presence (group C) or absence (group D) of embryos. These results demonstrated that completion of hatching was highest (52%; P less than 0.01) in group A after 6 days in culture. There were no significant differences between groups B, C and D (rates of total hatching 18, 17 and 17%, respectively). It is concluded that Vero cells improve the development of mouse embryos and this is likely to be due to removal of substances inhibitory to development.(ABSTRACT TRUNCATED AT 250 WORDS)
Semen parameters were correlated with the outcome of partial zona dissection (PZD) in 42 couples ... more Semen parameters were correlated with the outcome of partial zona dissection (PZD) in 42 couples with male factor infertility. Although fertilization rates were reduced, 12% of the embryos implanted following replacement. Spermatozoa from teratozoospermic sperm populations were able to fuse with oocytes following zona penetration through the artificial gaps. PZD followed by insemination with less than 5% normal spermatozoa led to 20 embryos which, upon replacement, did not implant. Motility and sperm count were not clearly correlated with the outcome of PZD and are therefore less useful indicators for patient selection. Teratozoospermic patients who previously failed to fertilize were compared to a group of similar patients who had not attempted IVF before. Although fertilization was significantly improved in first-time patients, 41% of the patients whose spermatozoa were initially unable to fertilize had at least one embryo when PZD was performed. Several pregnancies were established in this group. Subzonal sperm insertion (SZI) and PZD were compared in 19 patients using sibling oocytes. A significant fraction of spermatozoa from infertile men were able to fuse with the oolemma when directly inserted into the perivitelline area. Using a sucrose solution to shrink the ooplasm, only 1% of the oocytes were damaged during SZI. Monospermic fertilization rates following PZD and SZI were 15 and 16%, respectively. Both micromanipulation methods were successful in most patients. However, in two small groups of patients, only one technique resulted in fertilization.
The antibody to the rabbit sperm inner acrosomal membrane, raised in guinea pig, completely inhib... more The antibody to the rabbit sperm inner acrosomal membrane, raised in guinea pig, completely inhibited the fertilization of rabbit ova in vitro. The F(ab')2 of the antibody was equally effective in inhibiting fertilization. The antibody appeared to exert its inhibitory effect by binding to the inner acrosomal membrane of acrosome-reacted sperm. The antibody-treated sperm did not attach to or penetrate the zona pellucida. Thus, anti-IAM offers a great potential as a contraceptive agent.
Reproductive biology and endocrinology : RB&E, 2014
Polycystic ovarian syndrome (PCOS) is characterized by increased ovarian angiogenesis and vascula... more Polycystic ovarian syndrome (PCOS) is characterized by increased ovarian angiogenesis and vascularity. Accumulating evidence indicates that vascular endothelial growth factor (VEGF) is increased in PCOS and may play an important role in these vascular changes and the pathogenesis of this disease. Placental growth factor (PlGF), a VEGF family member, has not been previously characterized in PCOS women. We investigated levels and temporal expression patterns of PlGF and its soluble receptor sFlt-1 (soluble Fms-like tyrosine kinase) in serum and follicular fluid (FF) of women with PCOS during controlled ovarian stimulation. This was a prospective cohort study of 14 PCOS women (Rotterdam criteria) and 14 matched controls undergoing controlled ovarian stimulation. Serum was collected on day 3, day of hCG and day of oocyte retrieval. FF was collected on retrieval day. PlGF, sFlt-1 and anti-mullerian hormone (AMH) protein concentrations were measured using ELISA. Since sFlt-1 binds free Pl...
Polycystic ovarian syndrome (PCOS) ovaries are characterized by increased angiogenesis and hyperv... more Polycystic ovarian syndrome (PCOS) ovaries are characterized by increased angiogenesis and hypervascularity. While angiopoietin-1 (Ang-1) and its antagonist, angiopoietin-2 (Ang-2), are essential for ovarian function and angiogenesis, the levels of Ang-1 and Ang-2 in PCOS are unknown. This was a prospective cohort study of 14 PCOS women and 14 matched controls undergoing controlled ovarian stimulation (COS). Serum was collected on day 3, hCG and retrieval days. Follicular fluid (FF) was collected on retrieval day. Serum Ang-1 and Ang-2 levels were constant throughout COS, but serum Ang-1 levels were increased at all time points in PCOS women compared with controls (p < 0.05). No differences between groups were found in serum Ang-2 levels or FF Ang-1 levels. However, FF Ang-2 levels were increased almost 2-fold in PCOS women compared with controls (p < 0.01), and correlated positively with number of oocytes retrieved (r = 0.65, p < 0.0001). This study is the first to provide...
The Laxa group of the Panicum genus contains species which have CO(2) exchange and anatomical cha... more The Laxa group of the Panicum genus contains species which have CO(2) exchange and anatomical characteristics intermediate to C(3) and C(4) photosynthetic types (C(3)/C(4)), and also species characterized as C(3). Hybrids were made between two of the C(3)/C(4) species and two C(3) species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F(1) hybrids between Panicum milioides Nees. ex Trin (C(3)/C(4)) and P. laxum Mez. (C(3)), P. spathellosum Doell (C(3)/C(4)) and P. boliviense Hack. (C(3)), and P. spathellosum and P. laxum were studied. There were no consistent differences in apparent photosynthesis, although two of the three hybrids had higher net CO(2) uptake than the C(3) parent. Values of inhibition of apparent photosynthesis by 21% O(2), CO(2) loss in the light, and CO(2) compensation concentration for the hybrids were between those of the parents. All three hybrids showed leaf anatomical traits, especially organelle quantities in ...
Methods in molecular biology (Clifton, N.J.), 2004
ABSTRACT The cytoplasm of oocytes and early embryos is literally the “stuff of life.” The mature ... more ABSTRACT The cytoplasm of oocytes and early embryos is literally the “stuff of life.” The mature mammalian oocyte is an enormous cell invested with all the components required for the initiation of development. Through embryonic cloning scenarios, it is now clear that such cytoplasm is a primary requirement—necessary and sufficient (with some limitations) for supporting term development even through the control of a previously quiescent somatic genome (1,2). However, the components and mechanisms at work in the cytoplasm of eggs and early embryos satisfying this requirement remain mysterious and are the subject of considerable research. Ooplasm harbors the factors responsible for the events of fertilization/activation, maternal and paternal genome remodeling, epigenetic programming, embryonic genome activation and early development. Research into these areas is not only of great importance to our understanding of basic developmental biology, but also has critical implications in animal husbandry and biotechnology (3), as well as in assisted reproductive medicine (4). Experimental manipulation of the cytoplasm of oocytes and early embryos has been a significant component of this research. Also, such manipulation has recently been used to directly address ooplasm-specific defects in human reproduction (5). This chapter presents concepts and techniques for the manipulation of cytoplasm in mammalian eggs and early embryos and provides a brief review of knowledge gained from such research.
The role of FSH in inducing folliculogenesis is well established. Recently, the availability of p... more The role of FSH in inducing folliculogenesis is well established. Recently, the availability of pure FSH has led to a reevaluation of its role in the process of ovulation. Previously, these functions have been examined separately, usually with pregnant mares' serum gonadotropin (PMSG) followed by FSH for ovulation or FSH for folliculogenesis followed by hCG for ovulation. To determine if FSH alone can induce both folliculogenesis, ovulation and establish a functioning corpus luteum without exogenous LH, we injected sexually mature intact mice (CD-1) with either ovine FSH (oFSH, 5 micrograms; < 0.2% LH contamination) or recombinant FSH (RCFSH, 1 IU; devoid of any LH activity) to stimulate folliculogenesis, followed forty-eight hours later by a second injection of the same preparation (oFSH, 15 micrograms; RCFSH, 1 IU, respectively) to induce ovulation. Injected female mice were mated individually with a fertile male. On days 15-17, pregnancy rates and fetal development were ob...
Partial zona dissection (PZD) of human oocytes facilitates sperm penetration through mechanically... more Partial zona dissection (PZD) of human oocytes facilitates sperm penetration through mechanically made holes in the zona pellucida. Only 1 of 69 eggs was damaged when sucrose was used to shrink the ooplasm during micromanipulation. The fertilization rate of micromanipulated oocytes in 18 couples with male factor infertility was 68% (34/50), which compared favourably with insemination of non-micromanipulated controls (21/45, 47%). PZD was advantageous in oligozoospermic patients, but not in cases of asthenozoospermia, combined semen problems or immunological infertility. Three twin and two singleton pregnancies resulted following replacement of 23 micromanipulated and eight control embryos in 14 patients. No differences in embryo morphology and development rates were found between the micromanipulated and control groups. The incidence of polyspermy in couples with abnormal semen analyses was relatively low (less than 20%) possibly due to partial activation of the oocytes following exposure to sucrose. Polyspermy was high (57%) in normozoospermic patients with either immunological infertility (n = 3) or failure of fertilization in previous cycles (n = 4). In the three immunological patients, nine of 11 hyaluronidase and sucrose-exposed control embryos fertilized and six implanted, possibly indicating that cumulus and corona cells are contributing factors inhibiting fertilization in such cases.
Partial zona dissection (PZD), a method using mechanical force to open the human zona pellucida, ... more Partial zona dissection (PZD), a method using mechanical force to open the human zona pellucida, and zona drilling, which uses acidic Tyrode&#39;s (AT) medium, were compared in 1-day-old oocytes prior to reinsemination. The incidences of monospermy and polyspermy were 13/54 (24%) and 14/54 (26%) following PZD and 6/46 (13%) and 8/46 (17%) following the use of AT medium. This compared favorably with conventional reinsemination: 15/161 (9%) monospermy and 4/161 (3%) polyspermy. Three of the 27 PZD embryos became blastocysts, while none of the AT-exposed embryos developed satisfactorily. Eleven male-factor couples had some of their oocytes randomly treated with PZD prior to insemination; each of the patients had non-micromanipulated control oocytes. Monospermic fertilization and cleavage (23/34; 68%) doubled (P less than 0.05) when PZD was compared with the control oocytes (10/30; 33%). Replacing two PZD and a single control embryo in two patients resulted in twin pregnancies. A third twin pregnancy was established following replacement of only micromanipulated embryos.
... Jacques Cohen1*2, Carlene Eisner, Hilton Kort, Henry Malter2*3, Joe Massey, Mary Pat Mayer an... more ... Jacques Cohen1*2, Carlene Eisner, Hilton Kort, Henry Malter2*3, Joe Massey, Mary Pat Mayer and Klaus Wiemer ... In comparison, fertile couples with unrestricted intercourse are believed to conceive in 25% of menstrual cycles, when usually only one oocyte is released ...
Correction of polyspermy through pronucleus extraction in the absence of membrane relaxants was a... more Correction of polyspermy through pronucleus extraction in the absence of membrane relaxants was applied to 25 polyspermic human zygotes. Nine zygotes survived the procedure, and seven cleaved normally (two of which were fixed for chromosome analysis); two proceeded to compact and one cavitated. Eighteen of the polyspermic zygotes (14 with three pronuclei and 4 with four pronuclei) were obtained from zona pellucida-intact oocytes, and seven (1 five-pronucleate 3 four-pronucleate, and 3 three-pronucleate) from previously zona-drilled oocytes. Survival and cleavage occurred in all groups except in four- and five-pronucleate zona-drilled zygotes. Criteria used to identify male pronuclei were (1) pronucleus-associated sperm tails, (2) increased pronucleus size, and (3) greater distance (relative to female pronuclei) from the second polar body. Sperm tails were never seen and pronucleus size usually was identical. Therefore, the third criterion was used, although its reliability should be further evaluated. Until complete pronucleus removal techniques and reliable pronucleus selection criteria are perfected, embryo replacement after polyspermy correction could result in aneuploidy and molar pregnancy.
Zygotes from in vitro fertilization patients (n = 116) were randomly allocated to culture in eith... more Zygotes from in vitro fertilization patients (n = 116) were randomly allocated to culture in either conventional plastic petri dishes or coculture on a monolayer of fetal bovine uterine fibroblasts. Embryos (n = 288) remained 26 to 32 hours in these culture systems. Video tape recording for later morphological analysis (11 parameters) was performed on 117 conventionally cultured and 104 cocultured embryos, shortly before replacement, by an independent observer, unaware of the culture conditions for each embryo. A significantly greater number of cocultured embryos (52%) had &quot;good&quot; morphology (zero or only one abnormal characteristic) as compared with conventionally cultured embryos (30%). The most outstanding morphological characteristic of cocultured embryos was the expanded appearance of their blastomeres. The incidence of implantation per embryo increased from 13% to 19% when the coculture rather than conventional culture system was used, and the incidence of ongoing pregnancy per patient after coculture doubled to 35%.
To establish guidelines for application of partial zona dissection, subzonal sperm insertion, and... more To establish guidelines for application of partial zona dissection, subzonal sperm insertion, and regular in vitro fertilization (IVF) in severe male factor patients. Two studies were performed: partial zona dissection and IVF was applied in 57 couples during the first period, and subzonal sperm insertion was also applied in a second group of 47 couples. Procedures were performed in an academic research environment. Couples who failed fertilization previously, others not acceptable for IVF, and a third group in whom IVF was expected to fail. Oocytes were micromanipulated with either partial zona dissection or subzonal sperm insertion, or the zona pellucida was left intact. Embryos were replaced in patients prophylactically treated with methylprednisolone and antibiotics. Because several microsurgical fertilization techniques are now available, this study was performed to compare sperm parameters, embryo morphology, fertilization, and implantation rates after application of two successful micromanipulation procedures. Twenty-one pregnancies were established in 104 patients, 5 definitely from subzonal sperm insertion and 4 from partial zona dissection. Patients who failed IVF before had a similar chance of pregnancy after the use of micromanipulation, as first time patients (9/53 versus 12/51). In a subgroup of 15 patients who failed IVF with insufficient numbers of motile sperm, fertilization was significantly higher after subzonal sperm insertion. Partially zona-dissected embryos from couples with severe teratozoospermia (less than or equal to 5% normal forms; strict criteria) had significantly more morphological abnormalities than those from patients with moderate teratozoospermia (6% to 10% normal forms). In severely teratozoospermic patients, significantly fewer partially zona-dissected than subzonally inserted embryos implanted. The decision of which micromanipulation method to perform can possibly be based on careful analysis of sperm morphology.
Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embry... more Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embryos with Vero cells in both Dulbecco&#39;s modified Eagle&#39;s medium/Ham&#39;s F12 (DMEM/F12) and human tubal fluid (HTF) culture medium. Two-cell CB6F1 mouse embryos were cultured either in the presence of Vero cells (group A) or in culture medium alone (group B). In DMEM/F12 significantly more morulae developed in group A than in group B on day 3 (91 versus 23%; P less than 0.01). In contrast, the mouse embryos grew rapidly in HTF and significant differences were noted only in later embryonic stages (on day 5; 86% and 50%, P less than 0.01 of group A and B respectively, hatching or hatched). Similar experiments using DF1 and ICR mouse strains also revealed enhanced embryo development in the presence of Vero cells. To determine whether the embryo-enhancing effects of Vero cells were due to the removal of toxins or to the secretion of embryotrophic factors, ICR mouse embryos were cultured in fresh media with cells (group A), without cells (group B) and in cell-free conditions using cell-conditioned media which were obtained in the presence (group C) or absence (group D) of embryos. These results demonstrated that completion of hatching was highest (52%; P less than 0.01) in group A after 6 days in culture. There were no significant differences between groups B, C and D (rates of total hatching 18, 17 and 17%, respectively). It is concluded that Vero cells improve the development of mouse embryos and this is likely to be due to removal of substances inhibitory to development.(ABSTRACT TRUNCATED AT 250 WORDS)
Semen parameters were correlated with the outcome of partial zona dissection (PZD) in 42 couples ... more Semen parameters were correlated with the outcome of partial zona dissection (PZD) in 42 couples with male factor infertility. Although fertilization rates were reduced, 12% of the embryos implanted following replacement. Spermatozoa from teratozoospermic sperm populations were able to fuse with oocytes following zona penetration through the artificial gaps. PZD followed by insemination with less than 5% normal spermatozoa led to 20 embryos which, upon replacement, did not implant. Motility and sperm count were not clearly correlated with the outcome of PZD and are therefore less useful indicators for patient selection. Teratozoospermic patients who previously failed to fertilize were compared to a group of similar patients who had not attempted IVF before. Although fertilization was significantly improved in first-time patients, 41% of the patients whose spermatozoa were initially unable to fertilize had at least one embryo when PZD was performed. Several pregnancies were established in this group. Subzonal sperm insertion (SZI) and PZD were compared in 19 patients using sibling oocytes. A significant fraction of spermatozoa from infertile men were able to fuse with the oolemma when directly inserted into the perivitelline area. Using a sucrose solution to shrink the ooplasm, only 1% of the oocytes were damaged during SZI. Monospermic fertilization rates following PZD and SZI were 15 and 16%, respectively. Both micromanipulation methods were successful in most patients. However, in two small groups of patients, only one technique resulted in fertilization.
The antibody to the rabbit sperm inner acrosomal membrane, raised in guinea pig, completely inhib... more The antibody to the rabbit sperm inner acrosomal membrane, raised in guinea pig, completely inhibited the fertilization of rabbit ova in vitro. The F(ab')2 of the antibody was equally effective in inhibiting fertilization. The antibody appeared to exert its inhibitory effect by binding to the inner acrosomal membrane of acrosome-reacted sperm. The antibody-treated sperm did not attach to or penetrate the zona pellucida. Thus, anti-IAM offers a great potential as a contraceptive agent.
Reproductive biology and endocrinology : RB&E, 2014
Polycystic ovarian syndrome (PCOS) is characterized by increased ovarian angiogenesis and vascula... more Polycystic ovarian syndrome (PCOS) is characterized by increased ovarian angiogenesis and vascularity. Accumulating evidence indicates that vascular endothelial growth factor (VEGF) is increased in PCOS and may play an important role in these vascular changes and the pathogenesis of this disease. Placental growth factor (PlGF), a VEGF family member, has not been previously characterized in PCOS women. We investigated levels and temporal expression patterns of PlGF and its soluble receptor sFlt-1 (soluble Fms-like tyrosine kinase) in serum and follicular fluid (FF) of women with PCOS during controlled ovarian stimulation. This was a prospective cohort study of 14 PCOS women (Rotterdam criteria) and 14 matched controls undergoing controlled ovarian stimulation. Serum was collected on day 3, day of hCG and day of oocyte retrieval. FF was collected on retrieval day. PlGF, sFlt-1 and anti-mullerian hormone (AMH) protein concentrations were measured using ELISA. Since sFlt-1 binds free Pl...
Polycystic ovarian syndrome (PCOS) ovaries are characterized by increased angiogenesis and hyperv... more Polycystic ovarian syndrome (PCOS) ovaries are characterized by increased angiogenesis and hypervascularity. While angiopoietin-1 (Ang-1) and its antagonist, angiopoietin-2 (Ang-2), are essential for ovarian function and angiogenesis, the levels of Ang-1 and Ang-2 in PCOS are unknown. This was a prospective cohort study of 14 PCOS women and 14 matched controls undergoing controlled ovarian stimulation (COS). Serum was collected on day 3, hCG and retrieval days. Follicular fluid (FF) was collected on retrieval day. Serum Ang-1 and Ang-2 levels were constant throughout COS, but serum Ang-1 levels were increased at all time points in PCOS women compared with controls (p < 0.05). No differences between groups were found in serum Ang-2 levels or FF Ang-1 levels. However, FF Ang-2 levels were increased almost 2-fold in PCOS women compared with controls (p < 0.01), and correlated positively with number of oocytes retrieved (r = 0.65, p < 0.0001). This study is the first to provide...
The Laxa group of the Panicum genus contains species which have CO(2) exchange and anatomical cha... more The Laxa group of the Panicum genus contains species which have CO(2) exchange and anatomical characteristics intermediate to C(3) and C(4) photosynthetic types (C(3)/C(4)), and also species characterized as C(3). Hybrids were made between two of the C(3)/C(4) species and two C(3) species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F(1) hybrids between Panicum milioides Nees. ex Trin (C(3)/C(4)) and P. laxum Mez. (C(3)), P. spathellosum Doell (C(3)/C(4)) and P. boliviense Hack. (C(3)), and P. spathellosum and P. laxum were studied. There were no consistent differences in apparent photosynthesis, although two of the three hybrids had higher net CO(2) uptake than the C(3) parent. Values of inhibition of apparent photosynthesis by 21% O(2), CO(2) loss in the light, and CO(2) compensation concentration for the hybrids were between those of the parents. All three hybrids showed leaf anatomical traits, especially organelle quantities in ...
Methods in molecular biology (Clifton, N.J.), 2004
ABSTRACT The cytoplasm of oocytes and early embryos is literally the “stuff of life.” The mature ... more ABSTRACT The cytoplasm of oocytes and early embryos is literally the “stuff of life.” The mature mammalian oocyte is an enormous cell invested with all the components required for the initiation of development. Through embryonic cloning scenarios, it is now clear that such cytoplasm is a primary requirement—necessary and sufficient (with some limitations) for supporting term development even through the control of a previously quiescent somatic genome (1,2). However, the components and mechanisms at work in the cytoplasm of eggs and early embryos satisfying this requirement remain mysterious and are the subject of considerable research. Ooplasm harbors the factors responsible for the events of fertilization/activation, maternal and paternal genome remodeling, epigenetic programming, embryonic genome activation and early development. Research into these areas is not only of great importance to our understanding of basic developmental biology, but also has critical implications in animal husbandry and biotechnology (3), as well as in assisted reproductive medicine (4). Experimental manipulation of the cytoplasm of oocytes and early embryos has been a significant component of this research. Also, such manipulation has recently been used to directly address ooplasm-specific defects in human reproduction (5). This chapter presents concepts and techniques for the manipulation of cytoplasm in mammalian eggs and early embryos and provides a brief review of knowledge gained from such research.
The role of FSH in inducing folliculogenesis is well established. Recently, the availability of p... more The role of FSH in inducing folliculogenesis is well established. Recently, the availability of pure FSH has led to a reevaluation of its role in the process of ovulation. Previously, these functions have been examined separately, usually with pregnant mares' serum gonadotropin (PMSG) followed by FSH for ovulation or FSH for folliculogenesis followed by hCG for ovulation. To determine if FSH alone can induce both folliculogenesis, ovulation and establish a functioning corpus luteum without exogenous LH, we injected sexually mature intact mice (CD-1) with either ovine FSH (oFSH, 5 micrograms; < 0.2% LH contamination) or recombinant FSH (RCFSH, 1 IU; devoid of any LH activity) to stimulate folliculogenesis, followed forty-eight hours later by a second injection of the same preparation (oFSH, 15 micrograms; RCFSH, 1 IU, respectively) to induce ovulation. Injected female mice were mated individually with a fertile male. On days 15-17, pregnancy rates and fetal development were ob...
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