Sperm competition risk generates phenotypic plasticity in ovum fertilizability.

PubMed

Firman, Renée C; Simmons, Leigh W

2013-12-07

Theory predicts that sperm competition will generate sexual conflict that favours increased ovum defences against polyspermy. A recent study on house mice has shown that ovum resistance to fertilization coevolves in response to increased sperm fertilizing capacity. However, the capacity for the female gamete to adjust its fertilizability as a strategic response to sperm competition risk has never, to our knowledge, been studied. We sourced house mice (Mus domesticus) from natural populations that differ in the level of sperm competition and sperm fertilizing capacity, and manipulated the social experience of females during their sexual development to simulate conditions of either a future 'risk' or 'no risk' of sperm competition. Consistent with coevolutionary predictions, we found lower fertilization rates in ova produced by females from a high sperm competition population compared with ova from a low sperm competition population, indicating that these populations are divergent in the fertilizability of their ova. More importantly, females exposed to a 'risk' of sperm competition produced ova that had greater resistance to fertilization than ova produced by females reared in an environment with 'no risk'. Consequently, we show that variation in sperm competition risk during development generates phenotypic plasticity in ova fertilizability, which allows females to prepare for prevailing conditions during their reproductive life.

Estimating Young's modulus of zona pellucida by micropipette aspiration in combination with theoretical models of ovum

PubMed Central

Khalilian, Morteza; Navidbakhsh, Mahdi; Valojerdi, Mojtaba Rezazadeh; Chizari, Mahmoud; Yazdi, Poopak Eftekhari

2010-01-01

The zona pellucida (ZP) is the spherical layer that surrounds the mammalian oocyte. The physical hardness of this layer plays a crucial role in fertilization and is largely unknown because of the lack of appropriate measuring and modelling methods. The aim of this study is to measure the biomechanical properties of the ZP of human/mouse ovum and to test the hypothesis that Young's modulus of the ZP varies with fertilization. Young's moduli of ZP are determined before and after fertilization by using the micropipette aspiration technique, coupled with theoretical models of the oocyte as an elastic incompressible half-space (half-space model), an elastic compressible bilayer (layered model) or an elastic compressible shell (shell model). Comparison of the models shows that incorporation of the layered geometry of the ovum and the compressibility of the ZP in the layered and shell models may provide a means of more accurately characterizing ZP elasticity. Evaluation of results shows that although the results of the models are different, all confirm that the hardening of ZP will increase following fertilization. As can be seen, different choices of models and experimental parameters can affect the interpretation of experimental data and lead to differing mechanical properties. PMID:19828504

Transformation between scientific and social representations of conception: the method of serial reproduction.

PubMed

Bangerter, A

2000-12-01

The social representation (SR) of conception was investigated using an adapted version of Bartlett's (1932) method of serial reproduction. A sample of 75 participants reproduced a text describing the conception process in 20 segregated chains of four reproductive generations. Changes in sentence structure and content were analysed. Results indicated that when the scientific representation of conception is apprehended by laypersons, two different processes take place. First, the abstract biological description of the process is progressively transformed into an anthropomorphic description centred on the sperm and ovum (personification). Second, stereotypical sex-role attributes are projected onto the sperm and ovum. Limitations of the method of serial reproduction are discussed, as well as its potential for modelling processes of cultural diffusion of knowledge.

FINE STRUCTURE OF THE HUMAN OVUM IN THE PRONUCLEAR STAGE

PubMed Central

Zamboni, Luciano; Mishell, Daniel R.; Bell, James H.; Baca, Manuel

1966-01-01

A penetrated ovum was recovered from the oviduct of a 33 year old surgical patient who had had sexual intercourse 26 hr before the operation. The ovum was in the pronuclear stage. The ooplasmic organelles were mainly represented by mitochondria, endoplasmic reticulum components, and Golgi elements. Small vesicles were found in the space between the two sheets of the pronuclear envelope. These vesicles appeared to be morphologically similar to the ER vesicles in the ooplasm and were considered to be involved in pronuclear development. Numerous annulate lamellae were seen in the ooplasm as well as in the pronuclei. Ooplasmic crystalloids were also observed. These were thought to represent cytoplasmic yolk. Remnants of the penetrating spermatozoon were found in close relation to one of the pronuclei. The fine structure of the first and second polar body is also described. The nuclear complement of the first polar body consisted of isolated chromosomes, whereas the second polar body contained a membrane-bounded nucleus. In consideration of the possibility that polar body fertilization may take place, these differences in nuclear organization could be of importance. Other recognizable differences between the two polar bodies were presence of dense cortical granules and microvilli in the first polar body, and absence of these structures in the second. These dissimilarities were considered to be related to the organization of the egg cytoplasm at the time of polar body separation. PMID:6008199

Distribution, abundance, and predation effects of epipelagic ctenophores and jellyfish in the western Arctic Ocean

NASA Astrophysics Data System (ADS)

Purcell, Jennifer E.; Hopcroft, Russell R.; Kosobokova, Ksenia N.; Whitledge, Terry E.

2010-01-01

The Arctic Ocean is undergoing changes at an unprecedented rate because of global climate change. Especially poorly-studied in arctic waters are the gelatinous zooplankton, which are difficult to study using traditional oceanographic methods. A distinct zooplanktivore community was characterized in the surface 100 m by use of a Remotely Operated Vehicle, net collections, and SCUBA diving. The large scyphomedusa, Chrysaora melanaster, was associated with the warm Pacific water at ˜35-75 m depth. A diverse ctenophore community lived mainly above the C. melanaster layer, including Dryodora glandula, a specialized predator of larvaceans, Beroe cucumis, a predator of other ctenophores, and the extremely fragile Bolinopsis infundibulum, which was the most abundant species. Gut content analyses showed that Mertensia ovum selectively consumed the largest copepods ( Calanus spp.) and amphipods ( Parathemisto libellula); B. infundibulum consumed smaller copepods and pteropods ( Limacina helicina). Large copepods were digested by M. ovum in ˜12 h at -1.5 to 0 °C, but by B. infundibulum in only ˜4 h. We estimated that M. ovum consumed an average of ˜2% d -1 of the Calanus spp. copepods and that B. infundibulum consumed ˜4% d -1 of copepods <3 mm prosome length. These are significant consumption rates given that Calanus spp. have life-cycles of 2 or more years and are eaten by vertebrates including bowhead whales and arctic cod.

The discrimination of fish egg quality and viability by using Raman spectroscopy

NASA Astrophysics Data System (ADS)

Ishigaki, Mika; Sato, Hidetoshi

2014-03-01

Sexual reproductive body can be produced from a fertilized ovum. Once the ovum is fertilized with sperm, it runs through the cell division, differentiates to all kinds of cells, and goes to make a complete body. However, not all of them are viable and some of them stop to ontogenesis showing the developmental abnormality. In order to discriminate the egg quality, we apply Raman spectroscopy for fish egg. After the measurement, these Raman data are checked up with the information about the eggs can survive or not, and we examine what factors are important in egg components to distinguish between "good quality" and "not good quality". We present the results of assessment of egg quality, and investigate whether Raman spectroscopy can be used to a discriminate of egg quality.

Embryo quality is the main factor affecting cumulative live birth rate after elective single embryo transfer in fresh stimulation cycles.

PubMed

Niinimäki, Maarit; Veleva, Zdravka; Martikainen, Hannu

2015-11-01

The study was aimed to evaluate which factors affect the cumulative live birth rate after elective single embryo transfer in women younger than 36 years. Additionally, number of children in women with more than one delivery per ovum pick-up after fresh elective single embryo transfer and subsequent frozen embryo transfers was assessed. Retrospective cohort study analysing data of a university hospital's infertility clinic in 2001-2010. A total of 739 IVF/ICSI cycles with elective single embryo transfer were included. Analyses were made per ovum pick-up including fresh and subsequent frozen embryo transfers. Factors affecting cumulative live birth rates were examined in uni- and multivariate analyses. A secondary endpoint was the number of children born after all treatments. In the fresh cycles, the live birth rate was 29.2% and the cumulative live birth rate was 51.3%, with a twin rate of 3.4%. In the multivariate analysis, having two (odds ratio (OR) 1.73; 95% confidence interval (CI) 1.12-2.67) or ≥3 top embryos (OR 2.66; 95% CI 1.79-3.95) was associated with higher odds for live birth after fresh and frozen embryo cycles. Age, body mass index, duration of infertility, diagnosis or total gonadotropin dose were not associated with the cumulative live birth rate. In cycles with one top embryo, the cumulative live birth rate was 40.2%, whereas it was 64.1% in those with at least three top embryos. Of women who had a live birth in the fresh cycle, 20.4% had more than one child after all frozen embryo transfers. Among women with three or more top embryos after ovum pick-up, 16.1% gave birth to more than one child. The cumulative live birth rate in this age group varies from 40% to 64% and is dependent on the quality of embryos. Women with three or more top embryos have good chance of having more than one child per ovum pick-up without elevated risk of multiple pregnancies. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Reproductive biology of the Alaska skate Bathyraja parmifera, with comments on an intersexual individual.

PubMed

Matta, M E

2015-09-01

A total of 1357 specimens of Alaska skate Bathyraja parmifera were collected in the eastern Bering Sea by fisheries observers and during scientific groundfish surveys from 2003 to 2005. Male and female gonads were examined for maturity stage and seasonal reproductive timing. Based on seasonal reproductive data, including the occurrence of egg cases, ovum size, ovum number, shell-gland width and gonado-somatic index, this species appears to reproduce continually throughout the year. Potential effects of maternal size upon the size and number of mature oocytes were also investigated, with total length having a significant, although weak, influence on both. Morphology of a single intersexual individual encountered during the collection period is also described. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Dynamics of intracellular phospholipid membrane organization during oocyte maturation and successful vitrification of immature oocytes retrieved by ovum pick-up in cattle.

PubMed

Aono, Akira; Nagatomo, Hiroaki; Takuma, Tetsuya; Nonaka, Rika; Ono, Yoshitaka; Wada, Yasuhiko; Abe, Yasuyuki; Takahashi, Masashi; Watanabe, Tomomasa; Kawahara, Manabu

2013-05-01

The objective was to determine if immature bovine oocytes with cumulus cells at the germinal vesicle (GV) stage could be vitrified by aluminum sheets (AS; pieces of sheet-like aluminum foil). Cleavage rates in fertilized oocytes previously vitrified by the AS procedure were higher than those vitrified by a nylon-mesh holder (NM) procedure (89.3 ± 2.1% vs. 65.0 ± 3.7%). Cleaved embryos derived from the AS but not from the NM procedures developed to blastocysts. Furthermore, to investigate the effects of vitrifying GV oocytes on cytoplasmic structure and on the ability to undergo cytoplasmic changes, the intracellular phospholipid membrane (IM) was stained with the lipophilic fluorescent dye, 3,3'-dioctadecyloxa-carbocyanine perchlorate. After vitrification by AS, the IM remained intact relative to that of oocytes vitrified by NM. During in vitro maturation, reorganization of the IM was also undamaged in oocytes vitrified by AS before oocyte maturation, and the IM within oocytes vitrified by the NM procedure was evidently impaired. Finally, vitrification (AS) was used for GV oocytes collected using the ovum pick-up method. A bull calf was born after in vitro production and subsequent embryo transfer. The vitrification techniques described herein should facilitate generation of viable in vitro production bovine blastocysts using oocytes recovered using the ovum pick-up method. Copyright © 2013 Elsevier Inc. All rights reserved.

The status of assisted reproductive technology in Korea in 2012.

PubMed

Lee, Gyoung Hoon; Song, Hyun Jin; Choi, Young Min; Han, Hyuck Dong

2017-03-01

This study was designed to report the status of assisted reproductive technology (ART) therapy in South Korea between January 1, 2012 and December 31, 2012. A localized online survey, originally developed by the International Committee Monitoring Assisted Reproductive Technologies, was first launched and provided to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized as standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI. Thawed embryo transfer (TET) and other related procedures, including surgical sperm retrieval, were surveyed. Data from 33,956 ovum pick-up procedures were provided by 75 clinics in 2012. Of the 33,088 cycles in which ovums were retrieved, a complete transfer was performed in 90.5% (29,932 cycles). In addition, 10,079 FET cycles were confirmed to have resulted in clinical pregnancy, representing a pregnancy rate of 30.5% per ovum pick-up and 33.7% per ET. The most common number of embryos transferred in FET was 2 (41.6%), followed by 3 (34.0%), and non-elective single ETs (10.0%). Of the 10,404 TET cycles in which transfer was completed, 3,760 clinical pregnancies (36.1%) were confirmed by ultrasonography. The overall clinical pregnancy rate for FET and TET cycles in 2012 was higher than in 2011 (33.7% vs. 33.2% and 36.1% vs. 31.1%, respectively). The most common number of embryos transferred in FET cycles was 2, unlike in 2011.

The status of assisted reproductive technology in Korea in 2012

PubMed Central

Lee, Gyoung Hoon; Song, Hyun Jin; Han, Hyuck Dong

2017-01-01

Objective This study was designed to report the status of assisted reproductive technology (ART) therapy in South Korea between January 1, 2012 and December 31, 2012. Methods A localized online survey, originally developed by the International Committee Monitoring Assisted Reproductive Technologies, was first launched and provided to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized as standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI. Thawed embryo transfer (TET) and other related procedures, including surgical sperm retrieval, were surveyed. Results Data from 33,956 ovum pick-up procedures were provided by 75 clinics in 2012. Of the 33,088 cycles in which ovums were retrieved, a complete transfer was performed in 90.5% (29,932 cycles). In addition, 10,079 FET cycles were confirmed to have resulted in clinical pregnancy, representing a pregnancy rate of 30.5% per ovum pick-up and 33.7% per ET. The most common number of embryos transferred in FET was 2 (41.6%), followed by 3 (34.0%), and non-elective single ETs (10.0%). Of the 10,404 TET cycles in which transfer was completed, 3,760 clinical pregnancies (36.1%) were confirmed by ultrasonography. Conclusion The overall clinical pregnancy rate for FET and TET cycles in 2012 was higher than in 2011 (33.7% vs. 33.2% and 36.1% vs. 31.1%, respectively). The most common number of embryos transferred in FET cycles was 2, unlike in 2011. PMID:28428944

Fiber optic laser light scattering measurement of ciliary function of the fallopian tube

NASA Astrophysics Data System (ADS)

Halbert, Sheridan A.; Lim, Kap; Lee, Wylie I.

1990-07-01

A fiber-optic laser light-scattering system (FLS) for measuring ciliary function was evaluated by means of three sets of in vitro experiments. First, FLS performance was compared to that of a previously proven benchtop laser system (BLS). Using tissue excised from rabbit fallopian tubes, ciliary beat frequency (CBF) of each sample was measured with FLS and BLS. Paired CBF measurements showed excellent correlation between the two systems (r =0.93). Second, the FLS was used to evaluate the dependency of CBF on temperature (T) by using tissue sampies of rabbit oviductal fimbna. Regression analysis of CBF vs T showed a linear relationship over the range of 18-37°C for both individual samples (r =0.98) and pooled data from all experiments (r = 0.84). Fmally, the relalionship between CBF and ciliary ovum transport rate (TR) was tested by using T to modulate CBF of rabbit fimbria, in vitro. The relationship was linear over the range of CBF from 10 to 30 Hz (r2 = 0.83). At 37°C, CBF = 31+/-1 Hz, and TR = O.12+/-.02 mm/sec. equal to ovum transport rate in situ. The FLS is a valuable tool for characterizing ciliary activity and thus ovum transport function. Owing to the fact that ciliary dyskinesia resulting from disease of the fallopian tube is associated with infeitility, the FLS may be useful to acquire data important to the clinical evaluation of fallopian tube function and female infertility.

New Conceptions: Biosocial Innovations and the Family.

ERIC Educational Resources Information Center

Edwards, John N.

1991-01-01

Discusses asexual reproductive technologies (in vitro fertilization, cryopreservation, artificial insemination, sperm banks, genetic engineering, ovum transfer, cloning, ectogenesis, surrogacy) and suggests that theoretical implications of technologies may bring dramatic social changes in the family. Outlines symbolically representational…

In vitro oocyte maturation, fertilization and culture after ovum pick-up in an endangered gazelle (Gazella dama mhorr).

PubMed

Berlinguer, F; González, R; Succu, S; del Olmo, A; Garde, J J; Espeso, G; Gomendio, M; Ledda, S; Roldan, E R S

2008-02-01

The recovery of immature oocytes followed by in vitro maturation, fertilization and culture (IVMFC) allows the rescue of biological material of great genetic value for the establishment of genetic resource banks of endangered species. Studies exist on sperm cryopreservation of endangered Mohor gazelle (Gazella dama mhorr), but no work has been carried out yet on oocyte collection, fertilization and culture in this or related species. The purpose of this study was to develop a protocol for ovarian stimulation for the recovery of oocytes and subsequent IVMFC in the Mohor gazelle using frozen-thawed spermatozoa. Ovum pick-up was performed after ovarian stimulation with a total dose of 5.28 mg of ovine FSH. A total of 35 oocytes were recovered from 56 punctured follicles (62%) (N=6 females). Out of 29 cumulus-oocyte complexes matured in vitro, 3% were found at germinal vesicle stage, 7% at metaphase I, 21% were degenerated, and 69% advanced to metaphase II. Fertilization and cleavage rates of matured oocytes were 40 and 30%, respectively. Embryos cleaved in vitro up to the 6-8 cell stage but none progressed to the blastocyst stage, suggesting the existence of a developmental block and the need to improve culture conditions. Although more studies are needed to improve hormonal stimulation and oocyte harvesting, as well as IVMFC conditions, this study demonstrates for the first time the feasibility of in vitro fertilization with frozen-thawed semen of in vitro matured oocytes collected by ovum pick-up from FSH-stimulated endangered gazelles.

Short communication: Follicle superstimulation before ovum pick-up for in vitro embryo production in Holstein cows.

PubMed

Oliveira, Louise H; Sanches, Carlos P; Seddon, Adriano S; Veras, Marcio B; Lima, Flávio A; Monteiro, Pedro L J; Wiltbank, Milo C; Sartori, Roberto

2016-11-01

The objective was to evaluate in vitro embryo production (IVEP) in nonlactating Holstein cows after ovarian superstimulation. Cows were randomly assigned in a crossover design to 1 of 2 groups: control (n=35), which was not synchronized and not treated with hormones before ovum pick-up (OPU), or hormone-treated (n=35), in which wave emergence was synchronized and animals treated with porcine (p)-FSH in the presence of norgestomet before OPU. In the hormone-treated group, all follicles ≥7mm in diameter were aspirated for synchronization of wave emergence and cows received a norgestomet ear implant. After 36h, treatment with p-FSH (6 doses of 40mg each, 12h apart, i.m.) started. Ovum pick-up from follicles >2mm in diameter was performed 44h after the last p-FSH (coasting). Then, IVEP was performed. The total number of cumulus-oocyte complexes recovered (16.0 vs. 20.5±2.2) and number of grades I to III (viable) oocytes (10.7 vs. 12.3±1.6) did not differ between hormone-treated and control groups Additionally, no differences were found in the number of blastocysts per cow per OPU (3.0 vs. 2.6±0.5) or in blastocyst rates (17.1 vs. 12.2±2.4%) between hormone-treated and control, respectively. Thus, in this study, ovarian follicle superstimulation with p-FSH followed by coasting in nonlactating Holstein cows that had synchronization of wave emergence and progestin supplementation did not improve oocyte quality or IVEP compared to no hormonal treatment. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Early pregnancy factor (EPF) as a marker for the diagnosis of subclinical embryonic loss.

PubMed

Shahani, S K; Moniz, C; Chitlange, S; Meherji, P

1992-01-01

The validation of EPF as a possible correlate of early fertilization has made it possible to study and detect fertilization of the ovum in normal fertile women (during the luteal phase) and also in women with infertility, where the fertilization of the ovum may not be affected but there may be impairment in early embryonic development which results in early embryo loss or subclinical embryo loss. Our results have suggested that using EPF as a marker, we could detect subclinical embryonic loss in 57.8% of the infertile women where more than one menstrual cycle was studied and the blood was collected 4-7 days after ovulation. After the missed period, 80% of the patients who were negative for EPF but positive for hCG had spontaneous abortions. It would be interesting to study how EPF behaves as a marker, to detect subclinical embryonic loss in diverse pathological situations such as recurrent abortions, parental age and translocation carrier parents.

Reevaluating geographic variation in life-history traits of a widespread Nearctic amphibian

USGS Publications Warehouse

Davenport, Jon M.; Hossack, Blake R.

2016-01-01

Animals from cold environments are usually larger than animals from warm environments, which often produce clines in body size. Because variation in body size can lead to trade-offs between growth and reproduction, life-history traits should also vary across climatic gradients. To determine if life-history traits of wood frogs Rana sylvatica vary with climate, we examined female and male body length, clutch size, and ovum size from 37 locations across an unprecedented 32° of latitude. In conflict with recent research, body size, and ovum size decreased in cold climates and at higher latitudes. Clutch size did not vary with climate or latitude, but reproductive effort (clutch size:female size) did, suggesting selection for a life-history traits that favors maximizing propagule number over propagule size in cold climates. With accelerating climate change that will expose populations to novel environmental conditions, it is important to identify the limits of adaptation, which can be informed by greater understanding of variation in life-history traits.

Anti-Muellerian hormone levels in plasma of Holstein-Friesian heifers as a predictive parameter for ovum pick-up and embryo production outcomes

PubMed Central

VERNUNFT, Andreas; SCHWERHOFF, Mona; VIERGUTZ, Torsten; DIEDERICH, Mike; KUWER, Andreas

2014-01-01

The aim of this study was to investigate whether plasma anti-Muellerian hormone (AMH) levels of Holstein-Friesian heifers could be used to predict ovum pick-up (OPU) and embryo production outcomes. Plasma samples and data were collected from 64 heifers, which underwent repeated OPU with subsequent in vitro embryo production followed by embryo flushing after superovulation. AMH levels were significantly positively correlated with the number of follicles aspirated per OPU session (r = 0.45), recovered oocytes per OPU (r =0.43) and in vitro produced embryos per OPU (r = 0.28). No significant correlations between AMH and in vivo produced embryos were ascertained. Our results suggest that correlations between AMH and outcomes of an OPU-IVF program are too low to use AMH as a precise predictive parameter for the success of a particular OPU procedure in Holstein-Friesian heifers. However, AMH can help to identify groups of very good or very poor oocyte donors. PMID:25482112

Are All Highly Malignant Cancer Cells Identical?

DTIC Science & Technology

1979-01-01

embryo cells or even the original fertilized ovum. If this speculation has validity, the carcinogenesis and differentiation have the same destinies but...whose activity leads to the suppression of the transcrip- tion of the genes responsible for the unique set of embryo -cancer proteins and whose mutation

DYNAMIC BEHAVIOR OF A DELAY-DIFFERENTIAL EQUATION MODEL FOR THE HORMONAL REGULATION OF THE MENSTRUAL CYCLE

EPA Science Inventory

During the menstrual cycle, pituitary hormones stimulate the growth and development of ovarian follicles and the release of an ovum to be fertilized. The ovarian follicles secrete hormones during the cycle that regulate the production of the pituitary hormones creating positi...

Influence of Paternal Age on Assisted Reproduction Outcome

ClinicalTrials.gov

2017-04-27

We Will Retrospectively Assess Our Databases in Our Clinic; Instituto Valenciano de Infertilidad in Valencia (Spain); Searching for Assisted Reproduction Procedures; IUI Standard IVF/ICSI Cycles and Ovum Donation IVF/ICSI Cycles; Who Were Referred to Our Unit to Cryopreserve Sperm During the Period; From January 2000 to December 2006

Ova of Dioctophyme renale in a canine struvite urolith

PubMed Central

Whelen, Julie C.; Houston, Doreen M.; White, Carole; Favrin, Mike G.

2011-01-01

This report describes light and scanning electron microscopic appearance of an ovum from the giant kidney worm, Dioctophyme renale found in the nidus and stone layer of a struvite calculus. The stone had been surgically removed from the bladder of a 4-year-old spayed female mixed breed dog from northern Ontario. PMID:22654144

Factors affecting pregnancy rates after ovum pick up-derived embryo transfer in lactating Holstein recipients under tropical conditions

USDA-ARS?s Scientific Manuscript database

High milk production, heat, physiological status and management impair reproduction in Holstein cows. The use of in vivo-produced embryos has been reported as an alternative to enhance pregnancy outcome in the tropics; however there are several limitations for its production, especially from variati...

Improved embryo development in Japanese black cattle by in vitro fertilization using ovum pick-up plus intracytoplasmic sperm injection with dithiothreitol.

PubMed

Oikawa, Toshinori; Itahashi, Tomoko; Numabe, Takashi

2016-01-01

The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse, ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with low embryo production compared with the standard OPU-IVF.

Contributions of molecular size, charge distribution, and specific amino acids to the iron-binding capacity of sea cucumber (Stichopus japonicus) ovum hydrolysates.

PubMed

Sun, Na; Cui, Pengbo; Jin, Ziqi; Wu, Haitao; Wang, Yixing; Lin, Songyi

2017-09-01

This study investigated the contributions of molecular size, charge distribution and specific amino acids to the iron-binding capacity of sea cucumber (Stichopus japonicus) ovum hydrolysates (SCOHs), and further explored their iron-binding sites. It was demonstrated that enzyme type and degree of hydrolysis (DH) significantly influenced the iron-binding capacity of the SCOHs. The SCOHs produced by alcalase at a DH of 25.9% possessed the highest iron-binding capacity at 92.1%. As the hydrolysis time increased, the molecular size of the SCOHs decreased, the negative charges increased, and the hydrophilic amino acids were exposed to the surface, facilitating iron binding. Furthermore, the Fourier transform infrared spectra, combined with amino acid composition analysis, revealed that iron bound to the SCOHs primarily through interactions with carboxyl oxygen of Asp, guanidine nitrogen of Arg or nitrogen atoms in imidazole group of His. The formed SCOHs-iron complexes exhibited a fold and crystal structure with spherical particles. Copyright © 2017 Elsevier Ltd. All rights reserved.

Improved embryo development in Japanese black cattle by in vitro fertilization using ovum pick-up plus intracytoplasmic sperm injection with dithiothreitol

PubMed Central

OIKAWA, Toshinori; ITAHASHI, Tomoko; NUMABE, Takashi

2015-01-01

The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse, ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with low embryo production compared with the standard OPU-IVF. PMID:26460690

Science of breeding and heredity from ancient Persia to modern Iran.

PubMed

Kariminejad, Mohammad H; Khorshidian, Ardeshir

2012-01-01

About 1700 years BC, the prophet Zoroaster declared equal right for women and men to choose their "own ways." There is much evidence that ancient Persians believed in the equal contribution of women and men toward producing a child, and all its hereditary characteristics.Even more surprising are the phrases in Vandidad book, which were gathered by Mobedans in the Mad dynasty about egg extraction (gametes) from animal reproductive organs (gonads) and their storage for future conception.Centuries later, Western philosopher beliefs in regard to reproduction were contrary to Persian knowledge. The Greek philosophers believed that man's water (semen) contains all human characteristics, and the female uterus is only responsible for nurturing and development of fetus. After detection of the ovum (de Graaf 2(nd) half 17 century) Malpigy proposed the preformation theory (ovist) which means there is a miniature human inside ovum, that grows after Semen has entered the uterus and grow into a well-developed fetus. This hypothesis was later delegated to spermatozoa. These contradictory and inappropriate beliefs were subject to discussions and dispute, until C.E. Wolf demonstrated that the embryo is a product of the fertilization of ovum by spermatozoa.800 years prior this the sage Ferdowsi "The Great Iranian Poet" explains nicely the equal participation of man and woman in the production of the fetus and transmission of characters.After the renaissance and especially in recent years, tremendous achievements have been made in unraveling biological secrets of reproduction. There was no work o n genetics in Iran until 1936, when a genetic course was added to the biology curriculum in related colleges and universities; Iranian Genetics Society was founded in 1966, initiating a steady movement in this field.Although there was an inevitable gap during the revolution and war in our country, now there is great effort by researchers to eliminate the gap and bring us into the mainstream of world science, and development in biomedical sciences in the third millennium.

Theoretic Note: The Relation of Embryology to Linguistic and Cognitive Process

ERIC Educational Resources Information Center

Brown, Jason

2011-01-01

This paper is a conceptual essay that views the unfolding or individuation from the ovum to mind/brain form and process as the outcome of a unitary highly conserved pattern of epigenetic growth. The principle question concerns the extent to which the cognitive process can be understood as an extension or replication of primordial trends in the…

Effect of dietary marine lipids on female white bass ova compositions and progeny survival.

PubMed

Lewis, H A; Trushenski, J T; Lane, R L; Kohler, C C

2010-12-01

We evaluated white bass ovum fatty acid composition as well as embryonic and larval survival after varying n-3 and n-6 long-chain polyunsaturated fatty acid (LC-PUFA) concentrations in maternal diets. Diets containing graded levels (0, 33, 66, or 100%) of squid to menhaden oils were fed daily to apparent satiation to female white bass for 8 weeks prior to spawning. Embryonic survival was negatively related to maternal squid oil intake (P=0.015, R2=0.970). Squid oil-fed broodstock produced ova with decreased 20:5n-3 and increased C18 polyunsaturated fatty acid concentrations, largely reflecting the fatty acid profile of squid oil. Within ovum phospholipid, accumulation of 18:2n-6 may have altered biological function resulting in the lower embryonic survival among ova produced from the squid oil-fed broodstock. Our data suggest the importance of feeding white bass broodstock diets high in total n-3 LC-PUFA (at least 4.0% dry matter), and 20:5n-3-rich lipid sources such as menhaden oil can be effectively utilized by female white bass to produce quality ova.

Translation of globin messenger RNA by the mouse ovum

PubMed Central

Brinster, R. L.; Chen, H. Y.; Trumbauer, M. E.; Avarbock, M. R.

2016-01-01

It has been demonstrated that the Xenopus oocyte can translate rabbit haemoglobin messenger RNA (mRNA) following microinjection of the message into the cell1. The Xenopus oocyte has since been shown to be capable of translating a variety of messenger RNAs from different species2–4. This system has proved useful in understanding the mechanism of message translation and has also provided information about the translation capability of the Xenopus oocyte5,6. Several other cell types, including HeLa cells and fibroblasts, can also translate exogenous message injected into the cell7,8. However, there have been no reports of injection of mRNA into oocytes or fertilised one-cell ova of mammalian species. Nevertheless, the latter system could be of considerable use in studying the processing of exogenous messages in a mammalian system undergoing development, as well as providing insight into the way the early embryo processes injected messages and the protein products of such messages. We report here the results of injecting message into the fertilised one-cell mouse ovum and show that both mouse and rabbit globin mRNA are translated in this system. PMID:7352032

Scaphiopus couchii (Couch's spadefoot). Predation

USGS Publications Warehouse

Dayton, Gage H.; Jung, R.E.

1999-01-01

The observation described in this note appears to be the first record of ant predation upon anuran eggs. At an ephemeral pool in Big Bend National Park, Texas, we observed ants (Forelius mccooki) walking along a blade of grass onto the gelatin of a Couch?s spadefoot (Scaphiopus couchii) egg mass on the water surface. The ants had eaten through the gelatinous envelope and were harvesting the ovum and returning to their nest.

hCG Triggering in ART: An Evolutionary Concept.

PubMed

Hershko Klement, Anat; Shulman, Adrian

2017-05-17

Human chorionic gonadotropin (hCG) is no longer a single, omnipotent ovulation triggering option. Gonadotropin releasing hormone (GnRH) agonist, initially presented as a substitute for hCG, has led to a new era of administering GnRH agonist followed by hCG triggering. According to this new concept, GnRH agonist enables successful ovum maturation, while hCG supports the luteal phase and pregnancy until placental shift.

Ovarian endometriomas and IVF: a retrospective case-control study

PubMed Central

2011-01-01

We performed this retrospective case-control study analyzing 428 first-attempt in vitro fertilization (IVF) cycles, among which 254 involved women with a previous or present diagnosis of ovarian endometriosis. First, the results of these 254 cycles were compared with 174 cycles involving patients with proven non-endometriotic tubal infertility having similar age and body mass index. Women with ovarian endometriosis had a significantly higher cancellation rate, but similar pregnancy, implantation and delivery rates as patients with tubal infertility. Second, among the women with ovarian endometriosis, the women with a history of laparoscopic surgery for ovarian endometriomas prior to IVF and no visual endometriosis at ovum pick-up (n = 112) were compared with the non-operated women and visual endometriomas at ovum pick-up (n = 142). Patients who underwent ovarian surgery before IVF had significantly shorter period, lower antral follicle count and required higher gonadotropin doses than patients with non-operated endometriomas. The two groups of women with a previous or present ovarian endometriosis did, however, have similar pregnancy, implantation and live birth rates. In conclusion, ovarian endometriosis does not reduce IVF outcome compared with tubal factor. Furthermore, laparoscopic removal of endometriomas does not improve IVF results, but may cause a decrease of ovarian responsiveness to gonadotropins. PMID:21679474

Prepubertal propagation of transgenic cloned goats by laparoscopic ovum pick-up and in vitro embryo production.

PubMed

Baldassarre, H; Wang, B; Pierson, J; Neveu, N; Sneek, L; Lapointe, J; Cote, F; Kafidi, N; Keefer, C L; Lazaris, A; Karatzas, C N

2004-01-01

The use of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production was evaluated in the early propagation of cloned goats. Ten kinder goats produced by somatic cell nuclear transfer technology were used as oocyte donors. Half of the donor animals were subjected to LOPU at 2-3 months of age (prior to induction of lactation), whereas the other five goats were subjected to LOPU at 6-7 months of age (following induction to lactation). They were stimulated with 80 mg NIH-FSH-P1 (Folltropin, Vetrepharm, Canada) together with 300 IU eCG (Novormon, Vetrepharm, Canada) administered intramuscularly 36 h prior to LOPU. The number of follicles aspirated and oocytes recovered was higher in the younger group of donors (57 +/- 7 and 41 +/- 4 vs. 28 +/- 2 and 25.8 +/- 2, p < 0.05), however, oocytes from animals in the late prepubertal age showed higher developmental capacity resulting in higher transferable embryo yield (81.4% vs. 67.8%, p < 0.01), pregnancy rate (80% vs. 40%, p < 0.05) and total kids born (27 vs. 15, p < 0.01). In conclusion, LOPU in combination with in vitro embryo production techniques is an efficient method for the early propagation of valuable goats produced by somatic cell nuclear transfer.

Transcript map of the Ovum mutant (Om) locus: isolation by exon trapping of new candidate genes for the DDK syndrome.

PubMed

Le Bras, Stéphanie; Cohen-Tannoudji, Michel; Guyot, Valérie; Vandormael-Pournin, Sandrine; Coumailleau, Franck; Babinet, Charles; Baldacci, Patricia

2002-08-21

The DDK syndrome is defined as the embryonic lethality of F1 mouse embryos from crosses between DDK females and males from other strains (named hereafter as non-DDK strains). Genetically controlled by the Ovum mutant (Om) locus, it is due to a deleterious interaction between a maternal factor present in DDK oocytes and the non-DDK paternal pronucleus. Therefore, the DDK syndrome constitutes a unique genetic tool to study the crucial interactions that take place between the parental genomes and the egg cytoplasm during mammalian development. In this paper, we present an extensive analysis performed by exon trapping on the Om region. Twenty-seven trapped sequences were from genes in the databases: beta-adaptin, CCT zeta2, DNA LigaseIII, Notchless, Rad51l3 and Scya1. Twenty-eight other sequences presented similarities with expressed sequence tags and genomic sequences whereas 57 did not. The pattern of expression of 37 of these markers was established. Importantly, five of them are expressed in DDK oocytes and are candidate genes for the maternal factor, and 20 are candidate genes for the paternal factor since they are expressed in testis. This data is an important step towards identifying the genes responsible for the DDK syndrome.

Handbook of Human Tissue Sources. A National Resource of Human Tissue Samples

DTIC Science & Technology

1999-01-01

be frozen and thawed and still be viable for artificial insemination procedures or implan- tation. The newest type of human tissue storage for future...use is the storage of umbilical cord blood. SPERM, OVUM, AND EMBRYO BANKS Artificial insemination or donor insemination (DI) is a procedure to...anonymous human sperm for use in artificial insemination ; long-term semen storage for men facing the possibility of steril- ization, reduction in fertility

[Bioethical language in the law and jurisprudence about bioethical problems].

PubMed

Corral García, Eduardo

2013-01-01

The impact is analyzed that on the Spanish Law relative to questions bioethics--as the Law on artificial reproduction, the Law of biomedical investigation, and the Law on sexual and reproductive health--can have the conception of human embryo enunciated by the Court of Justice of the European Union in his judgment of October 18, 2011, considering it to be any ovum fertilized with independence of the degree of reached development.

Intrauterine instillation of diluted seminal plasma at oocyte pick-up does not increase the IVF pregnancy rate: a double-blind, placebo controlled, randomized study.

PubMed

von Wolff, M; Rösner, S; Germeyer, A; Jauckus, J; Griesinger, G; Strowitzki, T

2013-12-01

Does intrauterine application of diluted seminal plasma (SP) at the time of ovum pick-up improve the pregnancy rate by ≥14% in IVF treatment? Intrauterine instillation of diluted SP at the time of ovum pick-up is unlikely to increase the pregnancy rate by ≥14% in IVF. SP modulates endometrial function, and sexual intercourse around the time of embryo transfer has been suggested to increase the likelihood of pregnancy. A previous randomized double-blind pilot study demonstrated a strong trend towards increased pregnancy rates following the intracervical application of undiluted SP. As this study was not conclusive and as the finding could have been confounded by sexual intercourse, the intrauterine application of diluted SP was investigated in the present trial. A single-centre, prospective, double-blind, placebo-controlled, randomized, superiority trial on women undergoing IVF was conducted from April 2007 until February 2012 at the University Department of Gynaecological Endocrinology and Reproductive Medicine, Heidelberg, Germany. The study was powered to detect an 14% increase in the clinical pregnancy rate and two sequential tests were planned using the Pocock spending function. At the first interim analysis, 279 women had been randomly assigned to intrauterine diluted SP (20% SP in saline from the patients' partner) (n = 138) or placebo (n = 141) at the time of ovum pick-up. The clinical pregnancy rate per randomized patient was 37/138 (26.8%) in the SP group and 41/141 (29.1%) in the placebo group (difference: -2.3%, 95% confidence interval of the difference: -12.7 to +8.2%; P = 0.69). The live birth rate per randomized patient was 28/138 (20.3%) in the SP group and 33/141 (23.4%) in the placebo group (difference: -3.1%, 95% confidence interval of the difference: -12.7 to +6.6%; P = 0.56). It was decided to terminate the trial due to futility at the first interim analysis, at a conditional power of 62%. The confidence interval of the difference remains wide, thus clinically relevant differences cannot reliably be excluded based on this single study. The results of this study cast doubt on the validity of the concept that SP increases endometrial receptivity and thus implantation in humans. Funding was provided by the department's own research facilities. DRKS00004615.

Recurrent ectopic pregnancy in a woman suffering from infertility due to male factor presented with heterotopic pregnancy

PubMed Central

Chaudhari, Hemang D; Gandhi, Viplav S; Banker, Hiral; Suri, Amar

2012-01-01

Heterotopic pregnancy is the simultaneous occurrence of intrauterine and extrauterine pregnancies. A 27-year-old third gravida with history of two ectopic pregnancies, presented with cramping pain in pelvis radiating to left side and bleeding from vagina. Ultrasonographic diagnosis of heterotopic pregnancy was put forward. Unfortunately intrauterine component of heterotopic pregnancy resulted in blighted ovum and linear salpingostomy was done for left-sided tubal pregnancy. PMID:22605872

Ovum pick up, intracytoplasmic sperm injection and somatic cell nuclear transfer in cattle, buffalo and horses: from the research laboratory to clinical practice.

PubMed

Galli, Cesare; Duchi, Roberto; Colleoni, Silvia; Lagutina, Irina; Lazzari, Giovanna

2014-01-01

Assisted reproductive techniques developed for cattle in the last 25 years, like ovum pick up (OPU), intracytoplasmic sperm injection (ICSI), and somatic cell nuclear transfer, have been transferred and adapted to buffalo and horses. The successful clinical applications of these techniques require both the clinical skills specific to each animal species and an experienced laboratory team to support the in vitro phase of the work. In cattle, OPU can be considered a consolidated technology that is rapidly outpacing conventional superovulation for embryo transfer. In buffalo, OPU represents the only possibility for embryo production to advance the implementation of embryo-based biotechnologies in that industry, although it is still mainly in the developmental phase. In the horse, OPU is now an established procedure for breeding from infertile and sporting mares throughout the year. It requires ICSI that in the horse, contrary to what happens in cattle and buffalo, is very efficient and the only option because conventional IVF does not work. Somatic cell nuclear transfer is destined to fill a very small niche for generating animals of extremely high commercial value. The efficiency is low, but because normal animals can be generated it is likely that advancing our knowledge in that field might improve the technology and reduce its cost. Copyright © 2014 Elsevier Inc. All rights reserved.

Minimizing the Risk of Infection and Bleeding at Trans-Vaginal Ultrasound-Guided Ovum Pick-up: Results of a Prospective Web-Based World-Wide Survey.

PubMed

Bhandari, Harish; Agrawal, Rina; Weissman, Ariel; Shoham, Gon; Leong, Milton; Shoham, Zeev

2015-12-01

The objective of this study was to identify clinical practices worldwide, which would help in recognizing women at risk of excessive bleeding or of developing pelvic infection following trans-vaginal ovum pick-up (TV-OPU), measures taken to minimize risks and their management. A prospective, web-based questionnaire with distinct questions related to the practice of TV-OPU. A total of 155 units from 55 countries performing 97,200 IVF cycles annually responded to this web-based survey. A majority (65 %) responded that they would routinely carry out full blood count, while 35 % performed coagulation profile. Less than a third agreed screening women for vaginal infections. About a third used both sterile water and antiseptic to minimize ascending infection, and 52 % used antibiotics for prophylaxis. Doppler ultrasound was routinely used by 20 % of clinicians. 73 % of the clinicians preferred conservative management as their first line management for patients diagnosed with intra-abdominal bleeding. The study has identified a wide variation in the practices of minimizing infection and bleeding complications. The dearth of good quality evidence may be responsible for the lack of published guidelines, and therefore a lack of consensus on the optimum practice for minimizing the risk of infection and bleeding during TV-OPU.

Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Müllerian hormone profiles.

PubMed

Gamarra, G; Ponsart, C; Lacaze, S; Le Guienne, B; Humblot, P; Deloche, M-C; Monniaux, D; Ponter, A A

2015-11-01

Rapid genetic improvement in cattle requires the production of high numbers of embryos of excellent quality. Increasing circulating insulin and/or glucose concentrations improves ovarian follicular growth, which may improve the response to superovulation. The measurement of anti-Müllerian hormone (AMH) can help predict an animal's response to superovulation treatment. The aim of the present study was to investigate whether increasing circulating insulin concentrations, through propylene glycol (PG) drenches, could improve in vitro embryo production in oestrus-synchronised superovulated heifers with different AMH profiles. Holstein heifers were grouped according to pre-experimental AMH concentrations as low (L) or high (H). The PG drench increased circulating insulin and glucose concentrations and reduced β-hydroxybutyrate and urea concentrations compared with the control group. AMH was a good predictor of follicle and oocyte numbers at ovum pick-up (OPU), and of oocyte and embryo quality (AMH H>AMH L). PG in the AMH H group increased the number of follicles and blastocyst quality above that in the control group, but did not improve these parameters in the AMH L group. These results indicate that short-term oral PG supplementation modifies an animal's metabolic milieu and is effective in improving in vitro embryo production, after superovulation-OPU, more markedly in heifers with high rather than low AMH concentrations.

Evolution of egg target size: an analysis of selection on correlated characters.

PubMed

Podolsky, R D

2001-12-01

In broadcast-spawning marine organisms, chronic sperm limitation should select for traits that improve chances of sperm-egg contact. One mechanism may involve increasing the size of the physical or chemical target for sperm. However, models of fertilization kinetics predict that increasing egg size can reduce net zygote production due to an associated decline in fecundity. An alternate method for increasing physical target size is through addition of energetically inexpensive external structures, such as the jelly coats typical of eggs in species from several phyla. In selection experiments on eggs of the echinoid Dendraster excentricus, in which sperm was used as the agent of selection, eggs with larger overall targets were favored in fertilization. Actual shifts in target size following selection matched quantitative predictions of a model that assumed fertilization was proportional to target size. Jelly volume and ovum volume, two characters that contribute to target size, were correlated both within and among females. A cross-sectional analysis of selection partitioned the independent effects of these characters on fertilization success and showed that they experience similar direct selection pressures. Coupled with data on relative organic costs of the two materials, these results suggest that, under conditions where fertilization is limited by egg target size, selection should favor investment in low-cost accessory structures and may have a relatively weak effect on the evolution of ovum size.

Observations on the Biology of Afrotropical Hesperiidae (Lepidoptera) with particular reference to Kenya. Part 11. Heteropterinae.

PubMed

Cock, Matthew J W; Congdon, T Colin E

2017-01-30

Partial life histories from Kenya or Tanzania are presented for Metisella midas midas (Butler), M. medea medea Evans, M. orientalis orientalis Aurivillius, M. quadrisignatus nanda Evans, M. congdoni De Jong & Kielland and M. willemi Wallengren. The ovum of Metisella formosus linda Evans is also illustrated from Zambia. All feed on species of grasses (Poaceae). The convergence of the biology of the grass-feeding skippers, particularly Heteropterinae and Hesperiinae, Baorini is discussed.

Locating a modifier gene of Ovum mutant through crosses between DDK and C57BL/6J inbred strains in mice.

PubMed

Tan, Jing; Song, Gen Di; Song, Jia Sheng; Ren, Shi Hao; Li, Chun Li; Zheng, Zhen Yu; Zhao, Wei Dong

2016-06-01

A striking infertile phenotype has been discovered in the DDK strain of mouse. The DDK females are usually infertile when crossed with males of other inbred strains, whereas DDK males exhibit normal fertility in reciprocal crosses. This phenomenon is caused by mutation in the ovum (Om) locus on chromosome 11 and known as the DDK syndrome. Previously, some research groups reported that the embryonic mortality deviated from the semilethal rate in backcrosses between heterozygous (Om/+) females and males of other strains. This embryonic mortality exhibited an aggravated trend with increasing background genes of other strains. These results indicated that some modifier genes of Om were present in other strains. In the present study, a population of N₂2 (Om/+) females from the backcrosses between C57BL/6J (B6) and F₁ (B6♀ × DDK♂) was used to map potential modifier genes of Om. Quantitative trait locus showed that a major locus, namely Amom1 (aggravate modifier gene of Om 1), was located at the middle part of chromosome 9 in mice. The Amom1 could increase the expressivity of Om gene, thereby aggravating embryonic lethality when heterozygous (Om/+) females mated with males of B6 strain. Further, the 1.5 LOD-drop analysis indicated that the confidence interval was between 37.54 and 44.46 cM, ~6.92 cM. Amom1 is the first modifier gene of Om in the B6 background.

Characterization of sea cucumber (stichopus japonicus) ovum hydrolysates: calcium chelation, solubility and absorption into intestinal epithelial cells.

PubMed

Sun, Na; Cui, Pengbo; Lin, Songyi; Yu, Cuiping; Tang, Yue; Wei, Ye; Xiong, Youling; Wu, Haitao

2017-10-01

Sea cucumber (Stichopus japonicus) ovum hydrolysates (SCOHs) chelated with calcium were produced to investigate the characteristics of calcium binding and solubility, as well as to study any effects on calcium absorption by human intestinal epithelial cells. The results of the present study show that the calcium-binding capacity of SCOHs depended greatly on the type of proteases. The maximum level of Ca binding (0.38 mmol L -1 ) occurred when trypsin was used, with a peptide yield of 85.7%. Investigation of the possible chelating modes between SCOHs and calcium ions indicated that calcium ions bound to SCOHs primarily via interactions with carboxyl oxygen and amino nitrogen atoms of Glu and Asp and also that the phosphoserine residues might be also responsible for SCOH-calcium chelation. Moreover, SCOH-calcium complexes maintained the solubility of calcium under simulated gastrointestinal digestion, regardless of the presence of dietary components such as oxalate. Furthermore, SCOH-Ca led to higher peak intracellular [Ca 2+ ] i in both Caco-2 cells (338.3 nmol L -1 versus 269.6 nmol L -1 ) and HT-29 cells (373.9 nmol L -1 versus 271.7 nmol L -1 ) than casein phosphopeptide-Ca. Carboxyl oxygen and amino nitrogen atoms in the SCOHs could bind calcium ions, forming SCOH-calcium complexes. These complexes improved calcium solubility under simulated gastrointestinal digestion and also promoted calcium absorption in Caco-2 and HT-29 cells. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Phenotypic correlations between ovum pick-up in vitro production traits and pregnancy rates in Zebu cows.

PubMed

Vega, W H O; Quirino, C R; Serapião, R V; Oliveira, C S; Pacheco, A

2015-07-03

The growth of the Gyr breed in Brazil in terms of genetic gain for milk, along with conditions for market, has led to the use of ovum pick-up in vitro production (OPU-IVP) as a leader in biotechnology for the multiplication of genetic material. The aim of this study was to investigate phenotypic correlations between OPU-IVP-linked characteristics and pregnancy rates registered in an embryo transfer program using Gyr cows as oocyte donors. Data collected from 211 OPU sessions and 298 embryo transfers during the years 2012 and 2013 were analyzed and statistical analysis was performed. Estimates of simple Pearson correlations were calculated for NVcoc and PVcoc (number and proportion of viable cumulus-oocyte complexes, respectively); NcleavD4 and PcleavD4 (number and proportion of cleaved embryos on day 4 of culture, respectively); NTembD7 and PTembD7 (number and proportion of transferable embryos on day 7 of culture, respectively); NPrD30 and PPrD30 (number and proportion of pregnancies 30 days after transfer, respectively); and NPrD60 and PPrD60 (number and proportion of pregnancies 60 days after transfer, respectively). Moderate to moderately high correlations were found for all numerical characteristics, suggesting these as the most suitable parameters for selection of oocyte donors in Gyr programs. NVcoc is proposed as a selection trait due to positive correlations with percentage traits and pregnancy rates 30 and 60 days after transfer.

Oocyte recovery by ovum pick up and embryo production in river buffaloes (Bubalus bubalis).

PubMed

Manjunatha, B M; Ravindra, J P; Gupta, P S P; Devaraj, M; Nandi, S

2008-08-01

Ovum pick up (OPU) was conducted twice a week for 12 weeks in six cycling, non-descriptive (local breed), Indian buffaloes to study the efficiency of OPU on recovery of oocytes for embryo production. OPU was performed using an ultrasound equipment with a 5-MHz transvaginal transducer, a single-lumen, 18-gauge, 55-cm-long needle and a constant vacuum pressure of 110 mmHg. The number and size of follicles were determined before puncture. The recovered oocytes were graded, washed, matured for 24 h and then fertilized with frozen-thawed semen, followed by embryo culture on the oviductal monolayer. The mean number of follicles observed per animal per session did not differ between animals or between puncture sessions. A mean number of 3.62 +/- 0.32 mm follicles were observed, 2.90 +/- 0.15 mm follicles were punctured and 1.21 +/- 0.07 oocytes were recovered per animal per session, with an average recovery rate of 42%. Of the total oocytes recovered, 64% were suitable for in vitro embryo production (grade A + B) whereas 36% were classified to be of grades C + D. A mean number of 0.25 +/- 0.2 transferable embryos was produced in vitro per buffalo per session with a transferable embryo production rate of 32%. In conclusion, this study demonstrated that twice-a-week OPU could be applied repeatedly, without any adverse effects on the follicular growth and oocyte recovery and that recovered oocytes could be used for in vitro embryo production in buffaloes.

A new species of hydra (Cnidaria: Hydrozoa: Hydridae) and molecular phylogenetic analysis of six congeners from China.

PubMed

Wang, An-Tai; Deng, Li; Liu, Hong-Tao

2012-12-01

A new species of genus Hydra (Cnidaria: Hydrozoa: Hydridae), Hydra shenzhensis sp. nov. from Guangdong Province, China, is described and illustrated. Most polyps have five tentacles. Column length reaches 11 mm when relaxed. Buds do not acquire tentacles synchronously. Stenotele is broad and pyriform in shape, 1.2 times as long as its width. Holotrichous isorhiza is asymmetrical and slender (more than 2.7 times as long as its width), with transverse and slanting coils. Atrichous isorhiza is long, resembling a melon-seed in shape. Desmoneme is asymmetrically pyriform in shape. The new species, belonging to the vulgaris group, is dioecious; sexual reproduction was found to occur mostly during November and December under conditions of dense culture or food shortage. Two to thirteen testes, cone-like shape with papilla, formed beneath the tentacles. One to three ovaries, with an egg cup, milky white in color, formed on body column. Ninety percent of individuals developed only one ovum. On a mother polyp, a fertilized ovum developed an embryonic theca covering its surface. The embryotheca is brown, with a spine-like structure, covering a layer of transparent, membrane-like material. For phylogenetic analysis, the mitochondrial cytochrome oxidase subunit I gene (COI) of six hydra species collected from China was amplified by polymerase chain reaction (PCR) and sequenced. Morphological characters in combination with molecular evidence support the hydra described here as a new species.

Water conservation features of ova of Agrilus planipennis (Coleoptera: Buprestidae).

PubMed

Rigsby, Chad M; Cipollini, Don; Amstutz, Evan M; Smith, Terrance J; Yoder, Jay A

2013-04-01

The emerald ash borer, Agrilus planipennis Fairmaire, has destroyed millions of ash trees (Fraxinus spp.) in North America since first identified in Detroit in 2002. With species of ash distributed throughout North America, it is easy to speculate the extinction of all susceptible species of ash on the continent given a lack of physical, environmental, or climactic barrier for dispersal of the insect. We investigated water balance characteristics of emerald ash borer ova by using gravimetric methods in an effort to measure their response to heat- and water-stress and explore possible influences this stress may have on the ecology and physiology of the ovum. We also explored the possible water balance benefit of a peculiar, "clustering," oviposition behavior, as well as the difference in responses to stress between ova from a laboratory colony and ova from two wild populations. We found no evidence of water vapor absorption as a water balance strategy; rather enhanced water retention, resistance to desiccation, and viability with low water content were important survival strategies for these ova. Surface lipids resist thermal breakdown as indicated by ova having no detectable critical transition temperature, maintaining their water-proofing function as temperature rises. The observed "clustering" behavior had no desiccation-avoidance benefit and ova from the wild populations behaved almost identically to the ova from the lab colony, although the lab ova were slightly larger and more sensitive to dehydration. Given this new information, there appears to be no heat- or water-stress barriers for the dispersal of this devastating pest at the ovum stage.

Methylxanthines and reproduction.

PubMed

Minelli, Alba; Bellezza, Ilaria

2011-01-01

Reproduction is the process by which organisms create descendants. In human reproduction, two kinds of sex cells, or gametes, are involved. Sperm, the male gamete, and egg egg , or ovum ovum Vedi egg , the female gamete, must meet in the female reproductive system to create a new individual and both the female and the male reproductive systems are essential to the occurrence of reproduction. Scientific reports dealing with the effects of methylxanthines on reproduction are mostly centred on the use of these compounds as phosphodiesterase inhibitors that, by maintaining high intracellular levels of cyclic AMP (cAMP) cyclic AMP , will affect the gametes differently. High cAMP levels will sustain sperm sperm maturation while they hold the oocytes in mitotic arrest. Caffeine caffeine , being the methylxanthine most widely consumed by every segment of the population, has been the subject of greatest interest among health professionals and researchers. Conflicting results still seem to characterize the association between male/female caffeine caffeine consumption in adult life and semen quality/fertility fertility , although moderate daily caffeine consumption of levels up to 400-450 mg/day (5.7-6.4 mg/kg/day in a 70-kg adult) do not seem to be associated with adverse effects, i.e. general toxicity, effects on bone status and calcium balance, cardiovascular effects, behavioural changes, increased incidence of cancer, or effects on male fertility. A clear stimulation of egg-laying by the coffee leaf pest Leucoptera coffeella was recently reported, providing support for the hypothesis that caffeine, in a dose-dependent way, in insects stimulates egg-laying, thus leading to the death of coffee trees.

Quantification of hookworm ova from wastewater matrices using quantitative PCR.

PubMed

Gyawali, Pradip; Ahmed, Warish; Sidhu, Jatinder P; Jagals, Paul; Toze, Simon

2017-07-01

A quantitative PCR (qPCR) assay was used to quantify Ancylostoma caninum ova in wastewater and sludge samples. We estimated the average gene copy numbers for a single ovum using a mixed population of ova. The average gene copy numbers derived from the mixed population were used to estimate numbers of hookworm ova in A. caninum seeded and unseeded wastewater and sludge samples. The newly developed qPCR assay estimated an average of 3.7×10 3 gene copies per ovum, which was then validated by seeding known numbers of hookworm ova into treated wastewater. The qPCR estimated an average of (1.1±0.1), (8.6±2.9) and (67.3±10.4) ova for treated wastewater that was seeded with (1±0), (10±2) and (100±21) ova, respectively. The further application of the qPCR assay for the quantification of A. caninum ova was determined by seeding a known numbers of ova into the wastewater matrices. The qPCR results indicated that 50%, 90% and 67% of treated wastewater (1L), raw wastewater (1L) and sludge (~4g) samples had variable numbers of A. caninum gene copies. After conversion of the qPCR estimated gene copy numbers to ova for treated wastewater, raw wastewater, and sludge samples, had an average of 0.02, 1.24 and 67 ova, respectively. The result of this study indicated that qPCR can be used for the quantification of hookworm ova from wastewater and sludge samples; however, caution is advised in interpreting qPCR generated data for health risk assessment. Copyright © 2017. Published by Elsevier B.V.

Secondary SCNT doubles the pre-implantation development rate of reconstructed interspecies embryos by using cytoplasts of Sannen dairy goat ova.

PubMed

Zhang, Ai Min; Chen, Jian Quan; Sha, Hong Ying; Chen, Juan; Xu, Xu Jun; Wu, You Bin; Ge, Lai Xiang; Da, Hu Wei; Cheng, Guo Xiang

2007-10-01

The aim of this study was to investigate whether ova of Sannen goat could support the pre-implantation development of interspecies embryos constructed through somatic cell nucleus transfer (SCNT) embryos and whether secondary SCNT (SSCNT) could improve the pre-implantation development of those embryos. The primary SCNT (PSCNT) embryos were produced by using Sannen goat ovum cytoplasts as recipients and fibroblast cells, derived from human, rabbit and Boer goat skins, as nucleus donors. The blastomeres of 8 to 16 cells stage of PSCNT embryos were subsequently used as nucleus donor cells and Sannen goat ovum cytoplasts as recipients to evaluate the effect of SSCNT on the pre-implantation development rate of these reconstructed interspecies embryos. Our results indicate that the pre-implantation development rates of SSCNT embryos reconstructed using these three different blastomeres are almost twice of that of corresponding PSCNT embryos (human, 15.8% vs. 7.8%; rabbit, 27.9% vs. 12.5%; Boer goat 55.3% vs. 24.5%; P < 0.05 in all three cases). The time durations that embryos need for the serial events of remodeling and reprogramming to take place vary, depending on the animal species of nucleus donors. These data suggest that remodeling and reprogramming of donor nucleus may be enhanced by prolonged exposure of donor nucleus to maternal cytoplast. We conclude that Sannen goat cytoplast can support the pre-implantation development of embryos constructed with nuclei from various donors, including fibroblasts of human, rabbit and Boer goat; and the somatic nucleus derived from different species requires more time to achieve its reprogramming necessary for pre-implantation development.

Ovum pick up, in vitro embryo production, and pregnancy rates from a large-scale commercial program using Nelore cattle (Bos indicus) donors.

PubMed

Pontes, J H F; Melo Sterza, F A; Basso, A C; Ferreira, C R; Sanches, B V; Rubin, K C P; Seneda, M M

2011-06-01

The objective was to clarify in vitro production of bovine embryos in Brazil. Data from 656 ovum pick-up/in vitro production (OPU/IVP) procedures, performed on 317 Nelore (Bos indicus) donors, without hormone stimulation or control of ovarian follicular waves, were analysed. Donors were subjected to OPU from one to nine times (no specific schedule), with < 15 d between consecutive procedures. There were 20,848 oocytes, of which 15,747 (75.53%) were considered viable, 5,446 embryos were obtained, 5,398 embryos were immediately transferred, resulting in 1,974 pregnancies (36.57%) at Day 30 and 1,788 (33.12%) pregnancies at Day 60. The average number of total and viable oocytes produced per OPU session was (mean ± SEM) 30.84 ± 0.88 and 23.35 ± 0.7 (average of 8.1 ± 0.3 embryos and 3.0 ± 0.1 pregnancies per OPU-IVP procedure). Since oocyte production varied widely among donor, they were designated as very high, high, intermediate, and low, with 58.94 ± 2.04, 32.61 ± 0.50, 22.13 ± 0.50, and 10.26 ± 0.57 oocytes, respectively, produced by 78, 80, 79, and 80 donors. The number of viable oocytes recovered ranged from 0 to 128; since donors with numerous viable oocytes produced many viable embryos and pregnancies, oocyte production was useful for donor selection. However, there was no significant effect of the number of OPU sessions per donor on mean numbers of oocytes produced. In conclusion, we confirmed field reports of high oocyte production by some Nelore donors and demonstrated individual variation in oocyte yield, which was associated with embryo production and pregnancy rates. Copyright © 2011 Elsevier Inc. All rights reserved.

In vitro embryo production in buffalo (Bubalus bubalis) using sexed sperm and oocytes from ovum pick up.

PubMed

Liang, X W; Lu, Y Q; Chen, M T; Zhang, X F; Lu, S S; Zhang, M; Pang, C Y; Huang, F X; Lu, K H

2008-04-15

The objective was to explore the use of sexed sperm and OPU-derived oocytes in an IVP system to produce sex-preselected bubaline embryos. Oocytes were recovered from 20 fertile Murrah and Nili-Ravi buffalo cows by repeated (twice weekly) ultrasound-guided transvaginal ovum pick up (OPU), or by aspiration of abbatoir-derived bubaline ovaries, and subjected to IVF, using frozen-thawed sexed or unsexed bubaline semen. On average, 4.6 oocytes were retrieved per buffalo per session (70.9% were Grades A or B). Following IVF with sexed sperm, oocytes derived from OPU had similar developmental competence as those from abattoir-derived ovaries, in terms of cleavage rate (57.6 vs. 50.4%, P=0.357) and blastocyst development rate (16.0 vs. 23.9%, P=0.237). Furthermore, using frozen-thawed sexed versus unsexed semen did not affect rates of cleavage (50.5 vs. 50.9%, P=0.978) or blastocyst development (15.3 vs. 19.1%, P=0.291) after IVF using OPU-derived oocytes. Of the embryos produced in an OPU-IVP system, 9 of 34 sexed fresh embryos (26.5%) and 5 of 43 sexed frozen embryos (11.6%) transferred to recipients established pregnancies, whereas 7 of 26 unsexed fresh embryos (26.9%) and 6 out of 39 unsexed frozen embryos (15.4%) transferred to recipients established pregnancies. Eleven sex-preselected buffalo calves (10 females and one male) and 10 sexed buffalo calves (six females and four males) were born following embryo transfer. In the present study, OPU, sperm sexing technology, IVP, and embryo transfer, were used to produce sex-preselected buffalo calves. This study provided proof of concept for further research and wider field application of these technologies in buffalo.

Improvement of bovine in vitro embryo production by ovarian follicular wave synchronization prior to ovum pick-up.

PubMed

Cavalieri, F L B; Morotti, F; Seneda, M M; Colombo, A H B; Andreazzi, M A; Emanuelli, I P; Rigolon, L P

2018-09-01

This study evaluated the effects of the synchronization of ovarian follicular wave emergence on the efficiency of in vitro embryo production. Bos indicus cows (n = 20) were divided into two groups (control vs. synchronization) and subjected to repeated ovum pick-up (OPU) sessions (8 replicates each, with an interval of 21 days in a 2 × 2 crossover design) and subsequent in vitro embryo production. Cows in the control group (n = 10) were submitted to OPU procedures without any stimulation every 21 days. Animals in the synchronization group received a protocol-based progesterone implant, estradiol benzoate and prostaglandin on a random day of the estrus cycle (Day 0) and the OPU was performed on Day 5. After in vitro production, embryos were transferred to recipients synchronized at a fixed time and the diagnosis was performed 60 days later. An evaluation of the parameters for each OPU session revealed that donors that received the synchronization protocol pre-OPU showed a greater number of embryos (5.9 ± 0.5 vs. 4.5 ± 0.4; P = 0.037), higher rate of embryo production (45.8% vs. 38.5%; P = 0.001) and higher mean number of conceptions per group (2.2 ± 0.2 vs. 1.6 ± 0.2; P = 0.07) in relation to the group that did not receive hormonal treatment. We concluded that synchronization of the follicular wave prior to OPU showed positive effects on in vitro embryo production as well as on pregnancy rates. Copyright © 2017 Elsevier Inc. All rights reserved.

[Explore microcosmic connection between autophagy mechanism and follicular development based on "kidney governing reproduction" theory].

PubMed

Bai, Jun; Wu, Ke-Ming; Gao, Ran-Ran

2018-03-01

In the theory of traditional Chinese medicine(TCM) that "kidney storing essence and governing reproduction", reproductive essence is an important part of the kidney essence and acts as the original material of offspring embryos. Sperm, oocyte and zygote should be all included in the range of reproductive essence. Ovum is the essence of reproduction from inborn. The follicles maturation depends on the quality of oocyte and the vigor of kidney essence. Meanwhile, discharge of mature ovum relies on the stimulation and promotion by kidney Qi. Autophagy almost exists in different cells stages and all various of mammalian cells. Many studies have found that autophagy not only participates in the formation of follicles, but also in every phase of the follicles development, and is involved in the occurrence and development of ovarian diseases. Recently, more and more scholars believe that autophagy is a new field to explore the microcosmic relationship between autophagy and TCM. Kidney-nourishing TCM could promote follicular growth and improve variety clinical symptoms by inhibiting the apoptosis of ovarian granulosa cells and reducing follicular atresia. Meanwhile, apoptosis of ovarian granulosa cells is closely related to autophagy of ovarian granulosa cells. In order to provide some theoretical foundation for kidney-nourishing therapy's promoting effect on follicular growth and improving effect on ovarian function, also to further explore the molecular mechanism of kidney-nourishing medicine in promoting follicular development, this paper would explain the microcosmic relationship between autophagy and follicular development based on the theory of "kidney governing reproduction". All of these would be of great significance to prevent and intervene the diseases of reproductive system timely and effectively. Copyright© by the Chinese Pharmaceutical Association.

Cryopreservation of Living Organs

NASA Astrophysics Data System (ADS)

Tanasawa, Ichiro; Nagata, Shinichi; Kimura, Naohiro

Cryopreservation is considered to be the most promising way of preserving living organs or tissues for a long period of time without casuing any damage to their biological functions. However, cryopreservation has been succeeded only for simple and small-size tissues such as spermatozoon, ovum, erythrocyte, bone marrow and cornea. Cryopreservation of more complex and large-scale organs are not yet succssful. The authors have attempted to establish a technique for cryopreservation of larger living organs. An experiment was carried out using daphnia (water flea). The optimum rates of freezing and thawing were determined together with the optimum selection of cryoprotectant. High recovery rate was achieved under these conditions.

Environmental and epigenetic effects upon preimplantation embryo metabolism and development

PubMed Central

Chason, Rebecca J; Csokmay, John; Segars, James H.; DeCherney, Alan H.; Armant, D. Randall

2011-01-01

In vitro fertilization has provided a unique window into the metabolic processes that drive embryonic growth and development from a fertilized ovum to a competent blastocyst. Post-fertilization development is dependent upon a dramatic reshuffling of the parental genomes during meiosis, as well as epigenetic changes that provide a new and autonomous set of instructions to guide cellular differentiation both in the embryo and beyond. While early literature focused simply on the substrates and culture conditions required for progress through embryonic development, more recent insights lead us to suggest that the surrounding environment can alter the epigenome, which can, in turn, impact embryonic metabolism and developmental competence. PMID:21741268

Morphological diversity and evolution of egg and clutch structure in amphibians

USGS Publications Warehouse

Altig, Ronald; McDiarmid, Roy W.

2007-01-01

The first part of this synthesis summarizes the morphology of the jelly layers surrounding an amphibian ovum. We propose a standard terminology and discuss the evolution of jelly layers. The second part reviews the morphological diversity and arrangement of deposited eggs?the ovipositional mode; we recognize 5 morphological classes including 14 modes. We discuss some of the oviductal, ovipositional, and postovipositional events that contribute to these morphologies. We have incorporated data from taxa from throughout the world but recognize that other types will be discovered that may modify understanding of these modes. Finally, we discuss the evolutionary context of the diversity of clutch structure and present a first estimate of its evolution.

Early development and acquisition of Zooxanthellae in the temperate symbiotic sea anemone Anthopleura ballii (Cocks).

PubMed

Davy, Simon K; Turner, John R

2003-08-01

The ova of Anthopleura ballii become infected with zooxanthellae (endosymbiotic dinoflagellates) of maternal origin just prior to spawning. After fertilization, the zygotes undergo radial, holoblastic cleavage, and then gastrulate by invagination to form ciliated planulae. Because the zooxanthellae are localized on one side of the ovum-and later, within the blastomeres at one end of the embryo-invagination leads to the zooxanthellae being restricted to the planular endoderm and hence to the gastrodermal cells of the adult anemone. We propose that maternal inheritance of zooxanthellae plays an important part in the success of these temperate sea anemones, which live in regions where potential sources of zooxanthellae are scarce.

Ovum pick-up interval in buffalo (Bubalus bubalis) managed under wetland conditions in Argentina: Effect on follicular population, oocyte recovery, and in vitro embryo development.

PubMed

Konrad, J; Clérico, G; Garrido, M J; Taminelli, G; Yuponi, M; Yuponi, R; Crudeli, G; Sansinena, M

2017-08-01

The excellent adaptation of water buffalo (Bubalis bubalis) to swampy environments means that animals are frequently managed in areas with restricted access for reproductive procedures. The objective of this study was to evaluate the effect of the ovum pick-up (OPU) interval on follicular population, oocyte recovery, oocyte quality and in vitro embryo production. Twelve Murrah buffaloes were subjected to two consecutive dominant follicle reductions, and randomly assigned to either 7-day (n=6) or 14-day (n=6) OPU interval groups. Although there was no significant difference in the average number of small (<3mm) and large (>8mm) diameter follicles available per OPU, a higher proportion of medium-sized follicles (3-8mm) were observed in the 14-day interval group (5.129 vs 3.267; p<0.05). The number of recovered oocytes per donor was also significantly higher (4.51 vs. 2.8; p<0.05) in the 14-day interval group, although this was attributed to an increase in the proportion of lower quality oocytes (grades III and IV). After in vitro fertilization, embryo developmental competence from grade I and II oocytes was superior to that from grade III and IV oocytes, irrespective of OPU interval group. There was no significant difference in the proportion of grade I and II oocytes cleaved after sperm co-incubation; however, there was a higher proportion of blastocysts produced in 14-day interval group (28 vs. 6%, p<0.05). No blastocysts were produced from grade III and IV oocytes. This study indicates it is possible to use a 14-day interval for oocyte collection in water buffalo; this approach could be considered as an alternative when access to animals is restricted. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of eCG and FSH on ovarian response, recovery rate and number and quality of oocytes obtained by ovum pick-up in Holstein cows.

PubMed

Sendag, Sait; Cetin, Yunus; Alan, Muhammet; Hadeler, Klaus-Gerd; Niemann, Heiner

2008-06-01

The goal of the present study was to compare the ovarian response, oocyte yields per animal, and the morphological quality of oocytes collected by ultrasound guided follicular aspiration from Holstein cows treated either with FSH or eCG. Twenty four normal cyclic, German Holstein cows were randomly divided into two groups. Fourteen cows received 3000 IU eCG on day-4 prior to ovum pick-up (OPU) (day 0), 2 days later (day-2), 625 microg cloprostenol was administered. On day-1 GnRH was administered i.m. and 24h later OPU (day 0) was performed. In ten cows a total dose of 500 IU follicle stimulating hormone (Pluset) was administered intramuscularly in a constant dosage for 4 days with intervals of 12h, starting on day-5. Luteolysis was induced by application of 625 microg cloprostenol on day-2. On day-1 (24h after the last FSH treatment) GnRH was administered i.m. and 24h later OPU (day 0) was performed. Ovarian follicles were visualized on the ultrasound monitor, counted and recorded. All visible antral follicles were punctured. Recovered oocytes were graded morphologically based on the cumulus investment. Average follicle number in ovaries was higher in FSH group than eCG group (p<0.05). Oocyte yields per animal did not differ between FSH and eCG groups. The proportion of grade A oocytes was higher in the FSH group in the than eCG group (p<0.05). Likewise, rate of grade C oocytes in FSH group were lower than eCG group (p<0.05). In conclusion, these results suggest that ovarian response, follicle number in ovaries and oocyte quality are affected by the type of gonadotropin and FSH is better alternative than eCG for OPU treatment.

Ovarian follicular dynamics, follicle deviation, and oocyte yield in Gyr breed (Bos indicus) cows undergoing repeated ovum pick-up.

PubMed

Viana, J H M; Palhao, M P; Siqueira, L G B; Fonseca, J F; Camargo, L S A

2010-04-15

The objective of this study was to evaluate ovarian follicular dynamics during intervals between successive ovum pick-up (OPU) and determine its effects on the number and quality of recovered cumulus-oocyte complexes (COCs) in Zebu cows (Bos indicus). Pluriparous nonlactating Gyr cows (Bos indicus; n=10) underwent four consecutive OPU sessions at 96-h intervals. The dynamics of ovarian follicular growth between OPU sessions was monitored by twice-daily ultrasonographic examinations. A single dominant follicle (DF) or two codominant (CDF) follicles (>9mm) were present in 63.3% (19 of 30) of intervals studied, with follicle deviation beginning when the future dominant follicle (F1) achieved a diameter of 6.2+/-0.3mm. The phenomenon of codominance was observed in four (13.3%) of the inter-OPU intervals. The remaining intervals (36.6%, 11 of 30) were characterized by a greater follicular population, lower rate of follicular growth, and a smaller diameter F1 (P<0.0001). There was a tendency (P=0.08) toward an increase in the number of recovered COCs when dominant follicles were not present (NDF). The quality of COCs was not affected by the presence of a single dominant follicle, but codominant follicles resulted in recovery of a lower proportion of viable embryos (40.0%, 62.1%, and 63.6%; P<0.05) and higher proportions of degenerate COCs (56.0%, 30.3%, and 28.6%; P<0.05) for CDF, NDF, and DF respectively. We concluded that, in Zebu cows, (a) repeated follicle aspirations altered ovarian follicular dynamics, perhaps by increasing follicular growth rate; (b) follicular dominance could be established in cows undergoing twice-a-week OPU; and (c) the presence of a dominant follicle during short inter-OPU intervals may not affect COC quality, except when a codominant follicle was present. Copyright 2010 Elsevier Inc. All rights reserved.

Cryosurvival and pregnancy rates after exposure of IVF-derived Bos indicus embryos to forskolin before vitrification.

PubMed

Sanches, B V; Marinho, L S R; Filho, B D O; Pontes, J H F; Basso, A C; Meirinhos, M L G; Silva-Santos, K C; Ferreira, C R; Seneda, M M

2013-09-01

In vitro-produced (IVP) bovine embryos are more sensitive to cryopreservation than their in vivo counterparts due to their higher lipid concentrations, whereas Bos indicus IVP embryos are even more sensitive than Bos taurus IVP embryos. To examine the effects of a lipolytic agent, before vitrification of Bos indicus IVP embryos, on embryo survival, viability, and pregnancy rates, two experiments were conducted. In experiment 1, Bos indicus (Nelore) embryos were produced from abattoir-derived ovaries and allocated into two groups. In the treatment group, 10 μM of forskolin was added to the in vitro culture medium on Day 5 and incubated for 48 hours. On Day 7 of culture, IVP-expanded blastocysts from both the control (n = 101) and treatment (n = 112) groups were vitrified with ethylene glycol and DMSO via the Cryotop procedure. Although there was no significant difference between the rates of blastocoel reexpansion and hatching of the embryos exposed to forskolin (87.5% and 70.5%, respectively) compared with the control embryos (79.2% and 63.3%, respectively), the numerically superior rates of the embryos exposed to forskolin led to another experiment. In experiment 2, blastocysts produced from the ovum pick up were exposed or not exposed to the lipolytic agent and vitrified as in experiment 1. Embryos treated with forskolin had higher pregnancy rates than the control group (48.8% vs. 18.5%). In view of these results, 1908 Bos indicus embryos were produced from ovum pick up, exposed to the lipolytic agent, and blastocysts were transferred to recipients, and the pregnancy rates of the embryos of various breeds were compared. The mean pregnancy rate obtained was 43.2%. All data were analyzed by chi-square or by binary logistic regression (P ≤ 0.05). In conclusion, treatment with forskolin before vitrification improved cryotolerance of Bos indicus IVP embryos, resulting in good post-transfer pregnancy rates. Copyright © 2013 Elsevier Inc. All rights reserved.

[Effect of kuntai capsule on the number of retrieved oocytes, high-quality oocytes and embryos in in vitro fertilization of poor ovarian response patients].

PubMed

Lian, Fang; Jiang, Xiao-Yuan

2014-08-01

To observe the effect of Kuntai Capsule (KC) on the number of retrieved oocytes, the quality of high-quality oocytes and embryos in in vitro fertilization of poor ovarian response (POR) patients. Totally 70 POR patients preparing for in vitro fertilization-embryo transfer (IVF-ET) were randomly assigned to the observation group and the control group, 35 cases in each group. KC was administered to patients in the observation group in the preparation cycle (i.e., three menstrual cycles before IVF-ET) and during the superovulation process. Those in the control group took placebo during this period. Before and after medication the improvement of Shen yin deficiency syndrome (SYDS) was observed in the two groups. The basal follicle-stimulating hormone (bFSH), luteinizing hormone (LH), estradiol (E2), anti-Mullerian hormone (AMH), the ratio of FSH to LH, and antral follicle count (AFC) were observed. Besides, the E2 level of a single ovum on the day of HCG injection, the number of retrieved oocytes, the high-quality oocyte rate, and the high-quality embryos were observed. Compared with the control group, the SYDS, decreased bFSH and LH levels, increased ACF numbers, the E2 level of a single ovum on the day of HCG injection, the number of retrieved oocytes, high-quality oocytes, and high-quality embryos were superior in the observation group (P < 0.05). There was no statistical difference in the decreased FSH/LH level (P > 0.05). E2 and AMH increased after medication of KC in the observation group, while they decreased after administration of placebos in the control group. There was statistical difference in the post-pre treatment difference of E2 and AMH between the two groups (P < 0.05). KC could increase the number of retrieved oocytes, and elevate the quality of occytes and embryos in the IVF-ET.

Ovum pick up and in vitro production in the bovine after use in several generations: a 2005 status.

PubMed

van Wagtendonk-de Leeuw, A M

2006-03-15

The first In Vitro Produced (IVP) calf was born in 1981 and the non-surgical Ovum Pick Up (OPU) technique for the bovine was adapted from the human in 1987. Since then, considerable research has been aimed at improving both technologies in the bovine. Both OPU and IVP can now be seen as mature technologies. It can be estimated that more than 200,000 IVP calves have been born world wide to date, and when the two technologies are combined they are capable of producing over 50 calves per donor cow per year, albeit with a large variation between donors. Not many new breakthroughs are expected for OPU. For IVP however, automation and miniaturization as well as a greater understanding of the embryo through the application of gene based technologies such as micro-arrays, may provide an in vitro environment that is more in vivo-like than traditional micro drop/well systems. This improved environment should result in higher embryo developmental rates as well as improved quality and welfare of subsequent offspring. The application of OPU/IVP has progressed from treating infertile high genetic multiple ovulation and embryo transfer (MOET) cows in commercial situations to enhancing breeding scheme designs. With the bovine genome being rapidly sequenced and bovine genes for traits of economic interest becoming available in the coming years, OPU/IVP will prove invaluable in rapidly multiplying rare genes or Quantitative Trait Loci (QTL) of high value. In due course, it is anticipated that Marker Assisted Selection or Gene Assisted Selection (MAS/GAS) schemes will be more widely implemented. In addition, OPU, and particularly IVP, provide the basis for more advanced technologies such as cloning and transgenics. This paper is dedicated to celebrate and recognize the significant contributions made by Theo Kruip (1939-2003) to the wide area of bovine OPU and IVP.

The influence of the law on clinical decisions affecting life and death.

PubMed

Havard, J D

1983-07-01

The purpose of this lecture is to review the justification for legal interference in physicians' clinical decisions and the consequences of that interference to patients. Discussion covers contraception, abortion, negligence, and defensive medicine. Contraception is normally interpreted as the prevention or inhibition of fertilization or of implantation of the fertilized ovum in the uterus. The extra corporeal or in vitro fertilization program has raised the question of the legal and ethical status of the fertilized ovum before implantation. This is turn has raised questions about contraceptive devices or procedures whose purpose it is to prevent the implantation of a fertilized ovum, of which the IUd is the most common in use in the UK. Congressman Doonan of California moved to amend the US Constitution to provide that "life begins when a sperm cell fertilizes an egg." It may be assumed that this amendment is designed to prevent unacceptable experiments on spare embryos, but the consequences to family planning could be enormous unless some exception is included in favor of IUDs inserted for purposes of contraception. This leads to the issue of abortion and a reminder that until the 19th century it was not regarded as a crime in English common law to abort a fetus before "quickening" had occurred, as this was the point at which the embryo was regarded as having been animated. The Offenses Against the Person Act of 1861 established the current criminal offense of induced abortion to which the Abortion Act of 1967 now provides a defense. Recent developments in life support mechanisms have created difficulties over the extent to which such measures should be employed in the management of children born with life threatening abnormalities. A draft bill has been introduced requiring doctors to take all possible steps to feed defective neonates with life threatening abnormalities who are experiencing serious feeding problems. This means that these infants would have to be force fed irrespective of their prospects of survival and suffering which this will cause them and their parents. Clinical decisions have been most seriously affected by recent developments in the law of negligence. 1 of the main reasons for this has been the unsatisfactory way in which the adversary system of law ldeals with expert evidence. Attempts have been made to provide courts of law in the UK an agreed statement on expert medical matters, there is a long way to go before reaching the position achieved in many civil law countries on the continent of Europe where the experts recognized by the court hammer out an agreed upon opinion through scientific discourse and without the restrictions of evidentiary rules which are aimed more at the establishment of facts than the validity of scientific opinion. Those who attack the medical profession as being paternalistic and authoritative in making clinical decisions involving life and death fail to realize that the easy way out for the medical profession is to treat every case, however hopeless, with the full technology available, disregarding the patient's and family's interests, the costs in resources, and ignoring the stark reality of the problem.

Ensuring Quality of Experience in the Connected Home: A New Battleground for Quality of Experience Differentiation

NASA Astrophysics Data System (ADS)

Philpott, Michael; Doran, Jonathan

2016-03-01

The connected home is a critical part of the network-but one that has seen little in the way of true investment in recent years. With customer loyalty and churn directly linked to quality of experience, this is a part of the network that no players in the supply chain can afford to ignore any longer. Beyond broadband experience, the addition of smart technology to the connected home also represents one of the biggest revenue growth areas in the consumer telecoms market for some time. From both a churn-reduction and a revenue-growth perspective, investment in the connected home network must accelerate. This article outlines the key trends identified by Ovum in the connected home space for 2016 and beyond.

Paradoxical effects of bovine somatotropin treatment on the ovarian follicular population and in vitro embryo production of lactating buffalo donors submitted to ovum pick-up.

PubMed

Ferraz, M L; Sá Filho, M F; Batista, E O S; Watanabe, Y F; Watanabe, M R; Dayan, A; Joaquim, D C; Accorsi, M R; Gimenes, L U; Vieira, L M; Baruselli, P S

2015-03-01

The aim of the present study was to evaluate the effect of bovine somatotropin (bST; 500mg) administration on lactating buffalo donors submitted to two different ovum pick-up (OPU) and in vitro embryo production schemes with a 7 or 14d intersession OPU interval. A total of 16 lactating buffalo cows were randomly assigned into one of four experimental groups according to the bST treatment (bST or No-bST) and the OPU intersession interval (7 or 14d) in a 2×2 factorial design (16 weeks of OPU sessions). The females submitted to OPU every 14d had a larger (P<0.001) number of ovarian follicles suitable for puncture (15.6±0.7 vs. 12.8±0.4) and an increased (P=0.004) number of cumulus-oocyte complexes (COCs) recovered (10.0±0.5 vs. 8.5±0.3) compared to the 7d interval group. However, a 7 or 14d interval between OPU sessions had no effect (P=0.34) on the number of blastocysts produced per OPU (1.0±0.1 vs. 1.3±0.2, respectively). In addition, bST treatment increased (P<0.001) the number of ovarian follicles suitable for puncture (15.3±0.5 vs. 12.1±0.4) but reduced the percentage (18.9% vs. 10.9%; P=0.009) and the number (1.4±0.2 vs. 0.8±0.1; P=0.003) of blastocysts produced per OPU session compared with the non-bST-treated buffaloes. In conclusion, the 14d interval between OPU sessions and bST treatment efficiently increased the number of ovarian follicles suitable for puncture. However, the OPU session interval had no effect on embryo production, and bST treatment reduced the in vitro blastocyst outcomes in lactating buffalo donors. Copyright © 2015 Elsevier B.V. All rights reserved.

Epidemiology of double aneuploidies involving chromosome 21 and the sex chromosomes.

PubMed

Kovaleva, Natalia V; Mutton, David E

2005-04-01

The chance of two chromosome abnormalities occurring in one conceptus is very small. However, some authors have suggested that double aneuplodies (DAs) might be more common than the product of their individual frequencies. The nonrandomness of such DA events was considered to be evidence that nondisjunction (NDJ) may be genetically determined. Data collected from the National Down syndrome Cytogenetic Register (NDSCR) in England and Wales and from the literature indicate that the frequencies of all nonmosaic DAs, except for 48,XXY,+21, are lower than expected, probably because of strong intrauterine selection against such pregnancies. Collectively, we identified 52 cases of nonmosaic 48,XXY,+21; 28 cases of 48,XYY,+21; and 14 cases of 48,XXX,+21 in liveborns and 13 cases of 48,XXY,+21; four cases of 48,XYY,+21; and two cases of 48,XXX,+21 after prenatal diagnoses. Among these cases, analysis of the published unbiased cytogenetic surveys of liveborn DS revealed 24 cases of 48,XXY,+21; nine cases of 48,XYY,+21; and seven cases of 48,XXX,+21. These figures are different from the expected proportion of 1:1:1 (P < 0.001), with carriers of XXY overrepresented in the group of carriers of DA. Mechanisms put forth to account for the higher occurrence of 48,XXY,+21 may include greater accessibility of disomic ovum to Y-carrying sperm, and promotion of NDJ in ovum by Y-bearing sperm. 48,XXY,+21 DA was found to be age-dependent, as the proportion of mothers over age 35 (x = 33.0) was increased over the general population. This is in contrast to the apparently age-independent 48,XYY,+21 DA, with a mean maternal age of 24.7 (P < 0.001). Paternal ages were also remarkably different between the groups, with a mean age of 37.9 in 48,XXY,+21 cases and a mean age of 27.9 in 48,XYY,+21 cases (P < 0.01). Maternal age-related factors, rather than genetic predisposition, may play a more important role in the etiology of the most common DA, 48,XXY,+21.

Developmental competence of different quality bovine oocytes retrieved through ovum pick-up following in vitro maturation and fertilization.

PubMed

Saini, N; Singh, M K; Shah, S M; Singh, K P; Kaushik, R; Manik, R S; Singla, S K; Palta, P; Chauhan, M S

2015-12-01

In the present study, oocytes retrieved from cross bred Karan Fries cows by ovum pick-up technique were graded into Group 1 and Group 2, based on the morphological appearance of the individual cumulus-oocyte complexes (COCs). To analyze whether the developmental potential of the COCs bears a relation to morphological appearance, relative expression of a panel of genes associated with; (a) cumulus-oocyte interaction (Cx43, Cx37, GDF9 and BMP15), (b) fertilization (ZP2 and ZP3), (c) embryonic development (HSF1, ZAR1 and bFGF) and (d) apoptosis and survival (BAX, BID and BCL-XL, MCL-1, respectively) was studied at two stages: germinal vesicle (GV) stage and after in vitro maturation. The competence was further corroborated by evaluating the embryonic progression of the presumed zygotes obtained from fertilization of the graded COCs. The gene expression profile and development rate in pooled A and B grade (Group 1) COCs and pooled C and D grade (Group 2) COCs were determined and compared according to the original grades. The results of the study demonstrated that the morphologically characterized Group 2 COCs showed significantly (P<0.05) lower expression for most of the genes related to cumulus-oocyte interplay, fertilization and embryonic development, both at GV stage as well as after maturation. Group 1 COCs also showed greater expression of anti-apoptotic genes (BCL-XL and MCL1) both at GV stage and after maturation, while pro-apoptotic genes (BAX and BID) showed significantly (P<0.05) elevated expression in poor quality COCs at both the stages. The cleavage rate in Group 1 COCs was significantly higher than that of Group 2 (74.46±7.06 v. 31.57±5.32%). The development of the presumed zygotes in Group 2 oocytes proceeded up to 8- to 16-cell stages only, while in Group 1 it progressed up to morulae (35.38±7.11%) and blastocyst stages (9.70±3.15%), indicating their better developmental potential.

Causes of hatching failure in endangered birds

PubMed Central

Hemmings, N.; West, M.; Birkhead, T. R.

2012-01-01

About 10 per cent of birds' eggs fail to hatch, but the incidence of failure can be much higher in endangered species. Most studies fail to distinguish between infertility (due to a lack of sperm) and embryo mortality as the cause of hatching failure, yet doing so is crucial in order to understand the underlying problem. Using newly validated techniques to visualize sperm and embryonic tissue, we assessed the fertility status of unhatched eggs of five endangered species, including both wild and captive birds. All eggs were classified as ‘infertile’ when collected, but most were actually fertile with numerous sperm on the ovum. Eggs of captive birds had fewer sperm and were more likely to be infertile than those of wild birds. Our findings raise important questions regarding the management of captive breeding programmes. PMID:22977070

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mendizabal, A.F.; Quiroga, S.; Farinati, Z.

The usefulness of the direct 4-hour radioimmunoassay of estriol-16-glucuronide (E/sub 3/G) and pregnanediol-3-glucuronide (P/sub 2/G) in first morning urine (FMU) for establishing a prognosis of the early pregnancy outcome was evaluated in 106 patients that became pregnant. Microaliquots of FMU were serially assayed from day 3 of the conception cycle until day 80 of pregnancy. The E/sub 3/G and P/sub 2/G profiles of 19 pregnancies which terminated in spontaneous abortion with either a diagnosis of the blighted ovum syndrome (n = 11) or presumption of a corpus luteum/trophoblast failure (n = 8) have been compared with those of clinically normalmore » pregnancies (n = 87). Normal pregnancies displayed typical patterns of E/sub 3/G and P/sub 2/G development, while variations were observed in abortive events that reflected changes of the fetoplacental unit.« less

The baby broker boom.

PubMed

Annas, G J

1986-06-01

Annas comments on two 1986 court decisions involving surrogate motherhood: Surrogate Parenting Associates v. Kentucky and Smith v. Jones. In the first case, the Supreme Court of Kentucky ruled against the state's Attorney General in his attempt to revoke the charter of a company prompting surrogate arrangements. The court determined that the state's prohibition of child purchasing would not be violated if financial arrangements were worked out before conception and if the surrogate mother retained the right to cancel the contract up to the moment she relinquished her parental rights. In Smith v. Jones, a lower court judge in Michigan allowed an infertile ovum donor and her husband to be listed as the parents on a child's birth certificate, rather than the surrogate who had been artificially inseminated. Annas sees both decisions as accommodating the law to modern science, and as encouraging commercial surrogacy.

Surrogate motherhood revisited: maternal identity from a Jewish perspective.

PubMed

Jotkowitz, Alan

2011-12-01

A new bill regulating ovum donation in Israel is set to pass its second and third readings in the Israel Parliament in the upcoming months. The new law will expand the number of locally donated ova available, as previously Israeli women were prohibited from donating eggs unless they were undergoing fertility treatment. Parallel to this legislative initiative, there has been a change in rabbinical thinking over who is considered the mother in a case of surrogacy. Previously, the consensus has been that the birth mother is to be considered the mother, but over the last few years there has been a change in thinking and the genetic mother is now considered the mother. The purpose of this paper is to present the ethical and legal issues from a Jewish perspective in determining maternal identity. The dilemma also demonstrates some of the difficulties in applying Talmudic law to modern problems and the various methodologies used to overcome these issues.

Time-lapse cinematography of dynamic changes occurring during in vitro development of human embryos.

PubMed

Mio, Yasuyuki; Maeda, Kazuo

2008-12-01

The purpose of this study was to clarify developmental changes of early human embryos by using time-lapse cinematography (TLC). For human ova, fertilization and cleavage, development of the blastocyst, and hatching, as well as consequent changes were repeatedly photographed at intervals of 5-6 days by using an inverse microscope under stabilized temperature and pH. Photographs were taken at 30 frames per second and the movies were studied. Cinematography has increased our understanding of the morphologic mechanisms of fertilization, development, and behavior of early human embryos, and has identified the increased risk of monozygotic twin pregnancy based on prolonged incubation in vitro to the blastocyst stage. Using TLC, we observed the fertilization of an ovum by a single spermatozoon, followed by early cleavages, formation of the morula, blastocyst hatching, changes in the embryonic plates, and the development of monozygotic twins from the incubated blastocysts.

Upward swimming of a sperm cell in shear flow.

PubMed

Omori, Toshihiro; Ishikawa, Takuji

2016-03-01

Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.

Pregnancy rate and birth rate of calves from a large-scale IVF program using reverse-sorted semen in Bos indicus, Bos indicus-taurus, and Bos taurus cattle.

PubMed

Morotti, F; Sanches, B V; Pontes, J H F; Basso, A C; Siqueira, E R; Lisboa, L A; Seneda, M M

2014-03-15

Obtaining sexed sperm from previously frozen doses (reverse-sorted semen [RSS]) provides an important advantage because of the possibility of using the semen of bulls with desired genetic attributes that have died or have become infertile but from whom frozen semen is available. We report the efficiency of RSS on the pregnancy rate and birth rate of calves in a large-scale program using ovum pick-up and in vitro embryo production (IVEP) from Bos indicus, Bos indicus-taurus, and Bos taurus cattle. From 645 ovum pick-up procedures (Holstein, Gir, and Nelore), 9438 viable oocytes were recovered. A dose of frozen semen (Holstein, Nelore, Brahman, Gir, and Braford) was thawed, and the sperm were sex-sorted and cooled for use in IVF. Additionally, IVF with sperm from three Holstein bulls with freeze-thawed, sex-sorted (RSS) or sex-sorted, freeze-thawed (control) was tested. A total of 2729 embryos were produced, exhibiting a mean blastocyst rate of 29%. Heifers and cows selected for adequate body condition, estrus, and health received 2404 embryos, and 60 days later, a 41% average pregnancy rate was observed. A total of 966 calves were born, and 910 were of a predetermined sex, with an average of 94% accuracy in determining the sex. Despite the lower blastocyst rate with freeze-thawed, sex-sorted semen compared with sex-sorted semen, (P < 0.05), the pregnancy rate (bull I, 45% vs. 40%; II, 35% vs. 50%; and III, 47% vs. 48% for RSS and control, respectively; P > 0.05) and sex-sorted efficiency (bull I, 93% vs. 98%; II, 96% vs. 94%; and III, 96% vs. 97% for RSS and control, respectively; P > 0.05) were similar for each of the three bulls regardless of the sperm type used in the IVF. The sexing of previously frozen semen, associated with IVEP, produces viable embryos with a pregnancy rate of up to 40%, and calves of the desired sex are born even if the paternal bull has acquired some infertility, died, or is located a long distance from the sexing laboratory. Furthermore, these data show the feasibility of the process even when used in a large-scale IVEP program. Copyright © 2014 Elsevier Inc. All rights reserved.

Ovum pick-up in sheep: a comparison between different aspiration devices for optimal oocyte retrieval.

PubMed

Rodríguez, C; Anel, L; Alvarez, M; Anel, E; Boixo, J C; Chamorro, C A; de Paz, P

2006-04-01

In vivo ovum pick-up (OPU) in sheep may be improved with a proper choice of aspiration elements (needle and tubing) and aspiration vacuum pressure. In the present study, two experiments were carried out. In Expt 1, visible follicles in ovaries of slaughtered ewes (treated separately according to their diameters: small<3 mm, medium 3-5 mm and large>5 mm) were aspirated using different combinations of the three studied factors such as aspiration flow rate (10, 20, 30, 40 and 50 ml water/min), needle gauge (18 and 20 G) and tubing inner diameter (1, 2 or 3 mm internal diameter). In Expt 2, a study with two 18 G needles of different lengths (18 G: 82 mm; 18 GL: 600 mm) was carried out, using ovaries obtained post-mortem, and performing in vivo laparoscopic follicular aspiration on ewes. We considered good quality oocytes as those with both complete compact cumulus and a homogeneous cytoplasm. Recovery rate, proportion of good quality oocytes (good quality oocytes/100 oocytes recovered) and overall efficiency (good quality oocytes/100 follicles aspirated) were noted. In Expt 1, aspiration flow rate affect remarkable proportion of good quality oocytes (69.5%, 50.5%, 44.8%, 36.5% and 28.3% for flows from 10 to 50 ml/min respectively, p<0.05). Needle gauge did not affect aspiration device efficiency. Thin and intermediate tubings were more effective (overall efficiency rates: 34.9%, 32.3% and 28.1% for 1, 2 and 3 mm respectively, p<0.05). Follicle size did not affect recovery rate, but proportion of good quality oocytes was higher for large (77.9%) and medium (64.4%) follicles (p<0.05). Finally, some combinations of the aspiration device showed greater effectiveness. In Expt 2, needle length did not influence recovery rate, but good quality oocytes rate was significantly modified both post-mortem and in vivo (good quality rate for 18 G vs 18 GL needles: 69.5% vs 47.7% and 58.1% vs 25.4%, post-mortem and in vivo respectively, p<0.05). We conclude that low-aspiration flow rates (10 and 20 ml/min) with thin or intermediate tubings (1 and 2 mm), and any short needle (18 G or 20 G) are the most adequate aspiration factors for OPU in sheep.

Maternal influences on thiamine status of walleye Sander vitreus ova.

PubMed

Wiegand, M D; Johnston, T A; Brown, L R; Brown, S B; Casselman, J M; Leggett, W C

2011-03-01

Concentrations of the various forms of thiamine (vitamin B(1) ) were determined in walleye Sander vitreus ova from three central North American lakes. Total thiamine concentrations in ova from Lake Winnipeg S. vitreus were approximately three times greater (mean 12 nmol g(-1) ) than in those from Lakes Erie or Ontario. The percentage of thiamine in the active form (thiamine pyrophosphate, TPP) was highest in Lake Ontario ova (mean 88%) and lowest in those from Lake Winnipeg (mean 70%). Neither ova total thiamine concentration nor per cent ova thiamine as TPP showed any consistent relationships with maternal age, size, morphometric condition, somatic lipid concentrations or liver lipid concentrations. Ova total thiamine concentration, however, was negatively related to ovum size in some populations, as well as among populations, and was positively related to liver total thiamine concentration. Maternal transfer of thiamine to ova appears to be independent of female ontogenetic or conditional state in S. vitreus. © 2011 The Authors. Journal of Fish Biology © 2011 The Fisheries Society of the British Isles.

Unanswered questions on ectopic pregnancy.

PubMed

Jonas, E G

1980-07-19

In a previous article (3 May, p. 1127), the British Medical Journal attempted to assess the demography of ectopic pregnancy and noted that a rise in incidence might lead to a better diagnosis of the condition. Cited as possible causes of ectopic pregnancy are pelvic sepsis and IUD usage. There is clinical confirmation of the relationship between pelvic sepsis and IUD usage. A review of the records of 325 consecutive patients diagnosed as having ectopic pregnancy in 4 large London Hospitals during the period 1967-79 revealed that PID (Pelvic Inflammatory Disease) was uncommon (11%). 12% of the remaining patients had IUDs and a further 2% were progestogen-only contraceptive failures. As regards the role of IUDs in ectopic pregnancy, failed intrauterine contraception is hypothesized to result in pregnancy, but with an incidence of ectopic, mainly tubal, implantation by reasons of disturbed ovum migration along the oviduct. The physiology of the human oviduct is not well known. Further research should be done on the many common aberations of human reproduction, iatrogenic and spontaneous.

Comparison of techniques for the detection of helminth ova in drinking water and wastewater.

PubMed

Maya, C; Jimenez, B; Schwartzbrod, J

2006-02-01

Many countries use wastewater for irrigation. The World Health Organization established, as reuse guidelines, a maximum value of 1 helminth ovum/L for irrigation. Various techniques for enumerating helminth ova in water have been published. To determine the most adequate method for Mexico, four techniques were compared: the U.S. Environmental Protection Agency (U.S. EPA), membrane-filter, Leeds I, and Faust. Two types of water were used: drinking water and municipal wastewater effluent. Sensitivity, discrimination coefficients, precision, recovery efficiency, and cost were determined. In addition, several unseeded wastewater samples were analyzed. For drinking water, U.S. EPA and the membrane-filter techniques demonstrated comparable results; however, when wastewater was used, the membrane technique showed some deficiencies. Because the U.S. EPA technique may be used for samples with both high and low solids content, allows for the recovery of helminth ova with different specific gravities, and has the lowest total cost, it was selected as the best technique.

Rheotaxis facilitates upstream navigation of mammalian sperm cells

PubMed Central

Kantsler, Vasily; Dunkel, Jörn; Blayney, Martyn; Goldstein, Raymond E

2014-01-01

A major puzzle in biology is how mammalian sperm maintain the correct swimming direction during various phases of the sexual reproduction process. Whilst chemotaxis may dominate near the ovum, it is unclear which cues guide spermatozoa on their long journey towards the egg. Hypothesized mechanisms range from peristaltic pumping to temperature sensing and response to fluid flow variations (rheotaxis), but little is known quantitatively about them. We report the first quantitative study of mammalian sperm rheotaxis, using microfluidic devices to investigate systematically swimming of human and bull sperm over a range of physiologically relevant shear rates and viscosities. Our measurements show that the interplay of fluid shear, steric surface-interactions, and chirality of the flagellar beat leads to stable upstream spiralling motion of sperm cells, thus providing a generic and robust rectification mechanism to support mammalian fertilisation. A minimal mathematical model is presented that accounts quantitatively for the experimental observations. DOI: http://dx.doi.org/10.7554/eLife.02403.001 PMID:24867640

Major proteins of boar seminal plasma as a tool for biotechnological preservation of spermatozoa.

PubMed

Caballero, I; Vazquez, J M; García, E M; Parrilla, I; Roca, J; Calvete, J J; Sanz, L; Martínez, E A

2008-11-01

Boar seminal plasma is a complex mixture of secretions from the testes, epididymides, and the male accessory reproductive organs which bathe the spermatozoa at ejaculation. The seminal plasma contains factors, mostly proteins, which influence the spermatozoa, the female genital tract, and the ovum. In boars, most of the proteins belong to the spermadhesin family and bind to the sperm surface. Spermadhesins are multifunctional proteins with a wide range of ligand-binding abilities to heparin, phospholipids, protease inhibitors and carbohydrates; the family can be roughly divided into heparin-binding (AQN-1, AQN-3, AWN) and non-heparin-binding spermadhesins (PSP-I/PSP-II heterodimer). These proteins have various effects promoting or inhibiting sperm functions including motility, oviduct binding, zona binding/penetration, and ultimately fertilization. The complexity of the environmental signals that influence these actions have implications for the uses of these proteins in vivo and in vitro, and may lead to uses in improving sperm storage.

Development of insulin resistance in dairy cows by 150 days of lactation does not alter oocyte quality in smaller follicles.

PubMed

Oliveira, L H; Nascimento, A B; Monteiro, P L J; Guardieiro, M M; Wiltbank, M C; Sartori, R

2016-11-01

The objective of this study was to test the hypothesis that high-producing dairy cows become increasingly resistant to insulin throughout lactation and that, consequently, oocyte quality is compromised. We used Holstein cows at 50 (51.5±3.7; n=30), 100 (102.3±9.4; n=30), and 150 (154.5±18.9; n=30) days in milk (DIM). We measured circulating insulin and glucose and performed a glucose tolerance test (GTT) after 5h of fasting. To evaluate oocyte quality, we performed ovum pickup on the day before the GTT (581 oocytes). We performed statistical analyses using the MIXED procedure of SAS. The model included the fixed effects of DIM, period, time, parity, and an interaction between DIM and time. We observed no difference in the GTT between groups for any variable related to circulating glucose (for example, glucose peak=203.3±7.2, 208.8±6.3, and 194.3±5.9mg/dL). However, various measures of circulating insulin were different in cows at 150 DIM compared with 50 or 100 DIM: higher basal insulin (8.8±0.9, 8.8±0.8, and 11.9±0.8 µIU/mL), peak insulin (61.9±6.2 , 69.1±5.7, and 89.0±6.1 µIU/mL), delta maximum insulin (51.1±5.5 , 59.4±5.0, and 73.5±5.4 µIU/mL), and area under the curve 5-60 (1,874.8±171.0 , 2,189.5±157.8, and 2,610.5±174.0 µIU/mL × min). Nevertheless, we observed no difference among groups in the number of viable oocytes (3.2±0.7, 3.9±0.7, and 3.6±0.7 per cow per ovum pickup) or percentage of viable oocytes (49.3, 52.2, and 51.8%). Increased circulating insulin before and throughout the GTT in cows at 150 DIM indicates that cows develop increasing insulin resistance with increasing DIM; however, increased insulin resistance was not associated with a detectable alteration in the quality of oocytes aspirated from small and medium-sized follicles. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Beef donor cows with high number of retrieved COC produce more in vitro embryos compared with cows with low number of COC after repeated ovum pick-up sessions.

PubMed

Monteiro, F M; Batista, E O S; Vieira, L M; Bayeux, B M; Accorsi, M; Campanholi, S P; Dias, E A R; Souza, A H; Baruselli, P S

2017-03-01

The aim of the present study was to evaluate whether repeated ovum pick-up (OPU) procedures may affect the efficiency of OPU programs for in vitro embryo production (IVEP) in Bos indicus (Nelore) donors. In addition, the repeatability (r) efficiency of IVEP was also assessed. Data available were from 432 OPU-IVEP sessions that were performed at random stages of the estrous cycle in 36 cycling, nonlactating Nelore donors. Semen from three Nelore bulls was used for the IVF. Donors were submitted to 12 consecutive OPU procedures, with an interval of approximately 30 days between sessions. Data were analyzed as repeated measures using the GLIMMIX procedure of SAS 9.3. Cows yielding ≥15 COCs were defined as "high" COCs and cows with less than 15 COCs were defined as "low" COCs donors. The number of COCs retrieved decreased over time in donors classified with high COCs and remained fairly steady in cows with low COCs at the beginning of the program (P session*COC retrieved groups  = 0.02). Moreover, the number of COCs retrieved (P < 0.0001), and number of blastocysts produced per OPU (P = 0.001) was greater for the high COCs donors compared with the low COCs category, and these results were consistent across OPU sessions. Interestingly, there was no effect of COCs category on the rate of blastocyst development (P = 0.83). In addition, number of blastocysts produced were not affected by repeated OPU (P = 0.37) and interactions between time and COC category (P = 0.72). Similarly, blastocyst rate was not affected by repeated OPU (P = 0.21) and interactions between time and COC category (P = 0.58). Despite of COC category of the donor cows, repeatability was high for the number of COCs retrieved (r = 0.81), number of blastocysts produced per OPU (r = 0.79), and blastocyst rate (0.69). In conclusion, overall numbers of COCs decreased over time in donors classified as having high COCs. However, cumulative amounts of produced blastocysts were greater in donors with high COCs. More importantly, high repeatability was observed in terms of IVEP efficiency. Therefore, IVEP programs can be significantly improved with more aggressive selection toward donors with greater numbers of COCs. Copyright © 2016 Elsevier Inc. All rights reserved.

Superstimulation prior to the ovum pick-up to improve in vitro embryo production in lactating and non-lactating Holstein cows.

PubMed

Vieira, L M; Rodrigues, C A; Castro Netto, A; Guerreiro, B M; Silveira, C R A; Moreira, R J C; Sá Filho, M F; Bó, G A; Mapletoft, R J; Baruselli, P S

2014-07-15

The present study evaluated the efficacy of superstimulation with p-FSH (Folltropin) before the ovum pick-up (OPU) on IVP in lactating and nonlactating Holstein donors. A total of 30 Holstein cows (15 lactating and 15 nonlactating) were blocked by lactation status to one of two groups (control or p-FSH), in a cross-over design. On a random day of the estrous cycle, all cows received an intravaginal progesterone device and 2.0 mg IM of estradiol benzoate (Day 0). Cows in the control group received no further treatment, whereas cows in the p-FSH group received a total dosage of 200 mg of p-FSH on Days 4 and 5 in four decreasing doses 12 hours apart (57, 57, 43, and 43 mg). On Day 7, the progesterone device was removed, and OPU was conducted in both groups (40 hours after the last p-FSH injection in the p-FSH-treated group). There was no difference between groups (P = 0.92) in the numbers of follicles that were aspirated per OPU session (17.2 ± 1.3 vs. 17.1 ± 1.1 in control and p-FSH-treated cows, respectively); however, p-FSH-treated cows had a higher (P < 0.001) percentage of medium-sized follicles (6-10 mm) at the time of the OPU (55.1%; 285/517) than control cows (20.8%; 107/514). Although recovery rate was lower (60.0%, 310/517 vs. 69.8%, 359/514; P = 0.002), p-FSH-treated cows had a higher blastocyst production rate (34.5%, 89/258 vs. 19.8%, 55/278; P < 0.001) and more transferable embryos per OPU session were produced in the p-FSH group (3.0 ± 0.5 vs. 1.8 ± 0.4; P = 0.02). Regardless of treatment, non-lactating cows had a higher blastocyst rate (41.9%, 106/253 vs. 13.4%, 38/283; P = 0.001) and produced more transferable embryos per OPU session (3.5 ± 0.5 vs. 1.3 ± 0.3; P = 0.003) than lactating cows. Thus, superstimulation of Holstein donors with p-FSH before OPU increased the efficiency of IVP. In addition, non-lactating donors had higher percentage of in vitro blastocyst development and produced more embryos per OPU session than lactating cows. Copyright © 2014 Elsevier Inc. All rights reserved.

Long term effect of Ovum Pick-up in buffalo species.

PubMed

Neglia, Gianluca; Gasparrini, Bianca; Vecchio, Domenico; Boccia, Lucia; Varricchio, Ettore; Di Palo, Rossella; Zicarelli, Luigi; Campanile, Giuseppe

2011-02-01

The aim of this study was to evaluate the effect of an Ovum Pick-up (OPU) treatment carried out for 9 months in buffalo (Bubalus bubalis) species. Eight pluriparous non-lactating buffalo cows underwent OPU for 9 months. Recovered cumulus enclosed oocytes (COCs) were classified and COCs suitable for in vitro embryo production (IVEP) were in vitro matured (IVM), fertilized (IVF) and cultured (IVC) to the blastocyst (Bl) stage. Animals were monitored for a total period of 270 days, but at the summer solstice, follicular turnover decreased and at the 68-day of the trial, we decided to increase the OPU sampling interval from 3-4 to 7 days. It was therefore possible to distinguish two phases: a first phase (18 sessions), during which OPU was carried out twice weekly and a second phase (16 sessions) during which OPU sessions were performed weekly. This reduction did not modify the percentage of good quality COCs, while the incidence of grade D COCs decreased (P<0.01). Furthermore, embryo production was higher in the second phase, either if embryos were calculated on the total recovered COCs (8.3% vs. 21.4%; P<0.01) and on grade A+B COCs (13.0% vs. 32.1%; P<0.01), that supposedly should have given similar blastocyst yield. During the total period of the trial it was possible to distinguish a first period of 6 months (34 sessions) characterized by blastocyst production (0.36 blastocyst/buffalo/session), followed by an unproductive period of 3 months (12 sessions), during which embryos were not produced. During the first 6 months a higher (P<0.01) number of follicles (5.06 vs. 3.71), small follicles (3.38 vs. 2.07), total COCs (2.58 vs. 1.56) and good quality (A+B) COCs (1.51 vs. 0.94) per subject/session were recorded compared to the last 3 months. No Blastocyst were produced during the second period, even if the percentage of grade A+B COCs was similar to that recorded during the first period. In conclusion, buffalo cows submitted to repeated OPU sampling for a 9-month period, showed a decline of follicle recruitment and oocyte collection after the first two months of samplings. After 6-month of samplings, in spite of the quality grade of the collected oocytes, we found a drop in their developmental competence. Copyright © 2011 Elsevier B.V. All rights reserved.

Holographic diagnostics of biological microparticles

NASA Astrophysics Data System (ADS)

Dyomin, Victor V.; Sokolov, Vladimir V.

1996-05-01

Problem of studies of biological microojects is actual one for ecology, medicine, biology. Holographic techniques are useful to solve the problem. The above microojects are transparent or semitransparent ones in a visible light rather often. The case of an optically soft particle, (that is of a particle whose substance has the refractive index close to that of the surrounding medium) is quite probable in biological water suspensions. Some peculiarities of holographing optically soft microparticles are analyzed in this paper. We propose a technique to calculate a light intensity distribution in the plane of a hologram and in the plane of a holographic image of a particle of an arbitrary shape at an arbitrary distance from the latter plane. The efficiency of the approach proposed is demonstrated by calculational results obtained analytically for some simple cases. In a more complicated cases the technique can make a basis for numerical computations. The method of determining of refractive index of transparent and semitransparent microparticles is proposed. We also present in this paper some experimental results on holographic detection of the water drops and such optically soft particles as ovums of helmints in human jaundice.

Adrenomedullin increased the short-circuit current in the pig oviduct through chloride channels via the CGRP receptor: mediation by cAMP and calcium ions but not by nitric oxide.

PubMed

Liao, S B; Cheung, K H; Cheung, M P L; To, Y T; O, W S; Tang, F

2013-10-01

The oviduct serves as a site for the fertilization of the ovum and the transport of the conceptus down to the uterus for implantation. In this study, we investigated the presence of adrenomedullin (ADM) and its receptor component proteins in the pig oviduct. The effect of ADM on oviductal secretion, the specific receptor, and the mechanisms involved were also investigated. The presence of ADM and its receptor component proteins in the pig oviduct were confirmed using immunostaining. Short-circuit current (I(sc)) technique was employed to study chloride ion secretion in the oviductal epithelium. ADM increased I(sc) through cAMP- and calcium-activated chloride channels, and this effect could be inhibited by the CGRP receptor antagonist, hCGRP8-37. In contrast, the nitric oxide synthase inhibitor, L-NG-nitroarginine methyl ester (L-NAME), could not block the effect of ADM on I(sc). In summary, ADM may increase oviductal fluid secretion via chloride secretion independent of the nitric oxide pathway for the transport of sperm and the conceptus.

Differential expression of the metastasis suppressor KAI1 in decidual cells and trophoblast giant cells at the feto-maternal interface.

PubMed

Koo, Tae Bon; Han, Min-Su; Tadashi, Yamashita; Seong, Won Joon; Choi, Je-Yong

2013-10-01

Invasion of trophoblasts into maternal uterine tissue is essential for establishing mature feto-maternal circulation. The trophoblast invasion associated with placentation is similar to tumor invasion. In this study, we investigated the role of KAI1, an antimetastasis factor, at the maternal-fetal interface during placentation. Mouse embryos were obtained from gestational days 5.5 (E5.5) to E13.5. Immunohistochemical analysis revealed that KAI1 was expressed on decidual cells around the track made when a fertilized ovum invaded the endometrium, at days E5.5 and E7.5, and on trophoblast giant cells, along the central maternal artery of the placenta at E9.5. KAI1 in trophoblast giant cells was increased at E11.5, and then decreased at E13.5. Furthermore, KAI1 was upregulated during the forskolin-mediated trophoblastic differentiation of BeWo cells. Collectively, these results indicate that KAI1 is differentially expressed in decidual cells and trophoblasts at the maternal-fetal interface, suggesting that KAI1 prevents trophoblast invasion during placentation.

Immunocytogenetic effects of gonadotropin releasing hormone analogue: Triptorelin Pamoate (Decapeptyl) during in vitro fertilization treatment.

PubMed

Al-Qashi, S; Al-Qaoud, K M; Ja'fer, M; Khali, A M

2006-10-01

In this study, the immunocytogenetic effects of Decapeptyl (Triptorelin Pamoate) were assessed in the peripheral blood lymphocytes of females undergoing in vitro fertilization (IVF) treatment. Blood samples were taken from 34 females (23 treated and 11 controls), cultured and examined for sister chromatid exchanges (SCE) and cell replication index (CRI). The SCE frequency increased around ovulation time in the controls, and around the time of human chorionic gonadotropin administration in the IVF group. However, the SCE rate was significantly higher in the latter group. Furthermore, the white blood cells (WBC) count was significantly higher on the day of ovum pick up compared to the day preceding luteinizing hormone (LH) and follicle stimulating hormone (FSH) treatment. Similar observations were recorded with respect to phagocytic activity tested by nitroblue tetrazolium (NBT) assay. The nitric oxide production abilities of macrophages were not significantly changed in the LH, FSH-treated group relative to its control. Finally, the 50% complement hemolytic activity (CH50) assay results indicated that Decapeptyl lacks a significant potential to affect the complement system.

Assessment of sperm hyperactivated motility and acrosome reaction can discriminate the use of spermatozoa for conventional in vitro fertilisation or intracytoplasmic sperm injection: preliminary results.

PubMed

Wiser, A; Sachar, S; Ghetler, Y; Shulman, A; Breitbart, H

2014-04-01

Basic semen analysis is insufficient for determining the fertility potential. The aim of this study was to determine if hyperactivated motility (HAM) and acrosome reaction (AR) can be useful tests for evaluating semen quality during male infertility evaluations and to help the clinician decide whether regular insemination or intracytoplasmic sperm injection (ICSI) is preferable during in vitro fertilisation. A prospective study was conducted. Patients with normal sperm according to World Health Organization guidelines who underwent IVF treatment and planned regular insemination were asked to participate. A portion of sperm sample was evaluated for HAM and AR on day of ovum pick up. In HAM assessment, 93.3% of patients with increased HAM had a high fertilisation rate compared with 64% in the group without increased HAM (P = 0.059). For the AR evaluation, 91.7% of samples with a low rate of spontaneous AR had a high fertilisation rate compared with 39.3% in the group with a high rate of spontaneous AR (P = 0.004). © 2013 Blackwell Verlag GmbH.

[The desire for a child in today parenthood: Infertility and reproductive techniques].

PubMed

Alkolombre, Patricia

2014-01-01

The advances in reproductive medicine have unprecedentedly expanded the possibilities to form a family. Today we can speak of a diversity of ways to access to parenthood. The paradigm of family formations has also changed following the diversity of parenting projects: heterosexual couples, blended families (post-divorce), couples formed by lesbians and gays and single parenting. The vicissitudes of "desire for a child" under current parenthood lead us to think about their scope in clinical reproductive disorders or infertility and new family configurations. One of the destinies of the desire for a child in women is the "passion for a child", the child at any cost. Thinking of the changes in parenthood in this field involves not only the desire to have a child in a woman, a man or a couple, but also the history of the origins in children. Especially with some of the reproductive techniques used, such as surrogacy and the donation of gametes-eggs (ovum and sperm). In these cases we are dealing with new articulations of blood ties, kinship and parenthood.

Is there a 'new ethics of abortion'?

PubMed

Gillon, R

2001-10-01

This paper argues that the central issue in the abortion debate has not changed since 1967 when the English parliament enacted the Abortion Act. That central issue concerns the moral status of the human fetus. The debate here is not, it is argued, primarily a moral debate, but rather a metaphysical debate and/or a theological debate--though one with massive moral implications. It concerns the nature and attributes that an entity requires to have "full moral standing" or "moral inviolability" including a "right to life". It concerns the question when, in its development from newly fertilised ovum to unequivocally mature, autonomous morally inviolable person does a human being acquire that nature and those attributes, and thus a "right to life". The paper briefly reviews standard answers to these questions, outlining some problems associated with each. Finally there is a brief discussion of one way in which the abortion debate has changed since 1967--notably in the increasingly vociferous claim, especially from disability rights sectors, that abortion on grounds of fetal abnormality implies contempt for and rejection of disabled people--a claim that is rebutted.

Pregnancy outcome using general anesthesia versus spinal anesthesia for in vitro fertilization.

PubMed

Azmude, Azra; Agha'amou, Shahrzad; Yousefshahi, Fardin; Berjis, Katayoun; Mirmohammad'khani, Majid; Sadaat'ahmadi, Farahnaz; Ghods, Kamran; Dabbagh, Ali

2013-09-01

There is a considerable rate of fertility failure and this causes a great burden of untoward effects for patients. Usually a considerable number of these patients undergo anesthesia for their treatment. This study was designed to compare the effects of general and spinal anesthesia on these patients. In a randomized clinical trial, after taking informed written consent from the patients, 200 patients entered the study; 100 in each. During a 2 year period, women aged 20 to 40 years entered the study (one group receiving spinal anesthesia and the other, receiving general anesthesia). Ovum retrieval protocols were the same. Nonparametric and parametric analyses were used for data analysis. P value less than 0.05 was considered significant. There was no difference between the two groups regarding demographic variables. 15 of 100 patients (15%) in the general anesthesia group and 27 of 100 patients (27%) in the spinal anesthesia group had successful pregnancy after IVF; so, spinal anesthesia increased significantly the chance of IVF success (P value < 0.001; Chi Square). The results of this study demonstrated that spinal anesthesia increased the chance of fertilization success.

Analysis of intra-uterine fluid motion induced by uterine contractions.

PubMed

Eytan, O; Elad, D

1999-03-01

Evaluation of the fluid flow pattern in a non-pregnant uterus is important for understanding embryo transport in the uterus. Fertilization occurs in the fallopian tube and the embryo (fertilized ovum) enters the uterine cavity within 3 days of ovulation. In the uterus, the embryo is conveyed by the uterine fluid for another 3 to 4 days to a successful implantation site at the upper part of the uterus. Fluid movements within the uterus may be induced by several mechanisms, but they seem to be dominated by myometrial contractions. Intra-uterine fluid transport in a sagittal cross-section of the uterus was simulated by a model of wall-induced fluid motion within a two-dimensional channel. The time-dependent fluid pattern was studied by employing the lubrication theory. A comprehensive analysis of peristaltic transport resulting from symmetric and asymmetric contractions is presented for various displacement waves on the channel walls. The results provide information on the flow field and possible trajectories by which an embryo may be transported before implantation at the uterine wall.

Nonpermissiveness for mouse embryonic stem (ES) cell derivation circumvented by a single backcross to 129/Sv strain: establishment of ES cell lines bearing the Omd conditional lethal mutation.

PubMed

Kress, C; Vandormael-Pournin, S; Baldacci, P; Cohen-Tannoudji, M; Babinet, C

1998-12-01

The inbred mouse strain DDK carries a conditional early embryonic lethal mutation that is manifested when DDK females are crossed to males of other inbred strains but not in the corresponding reciprocal crosses. It has been shown that embryonic lethality could be assigned to a single genetic locus called Ovum mutant (Om), on Chromosome (Chr) 11 near Syca 1. In the course of our study of the molecular mechanisms underlying the embryonic lethality, we were interested in deriving an embryonic stem cell bearing the Om mutation in the homozygous state (Omd/Omd). However, it turned out that DDK is nonpermissive for ES cell establishment, with a standard protocol. Here we show that permissiveness could be obtained using Omd/Omd blastocysts with a 75% 129/Sv and 25% DDK genetic background. Several germline-competent Omd/Omd ES cell lines have been derived from blastocysts of this genotype. Such a scenario could be extended to the generation of ES cell lines bearing any mutation present in an otherwise nonpermissive mouse strain.

In vitro embryo production in goats: Slaughterhouse and laparoscopic ovum pick up-derived oocytes have different kinetics and requirements regarding maturation media.

PubMed

Souza-Fabjan, Joanna Maria G; Locatelli, Yann; Duffard, Nicolas; Corbin, Emilie; Touzé, Jean-Luc; Perreau, Christine; Beckers, Jean François; Freitas, Vicente José F; Mermillod, Pascal

2014-05-01

A total of 3427 goat oocytes were used in this study to identify possible differences during in vitro embryo production from slaughterhouse or laparoscopic ovum pick up (LOPU) oocytes. In experiment 1, one complex, one semi-defined, and one simplified IVM media were compared using slaughterhouse oocytes. In experiment 2, we checked the effect of oocyte origin (slaughterhouse or LOPU) on the kinetics of maturation (18 vs. 22 vs. 26 hours) when submitted to semi-defined or simplified media. In experiment 3, we determined the differences in embryo development between slaughterhouse and LOPU oocytes when submitted to both media and then to IVF or parthenogenetic activation (PA). Embryos from all groups were vitrified, and their viability evaluated in vitro after thawing. In experiment 1, no difference (P > 0.05) was detected among treatments for maturation rate (metaphase II [MII]; 88% on average), cleavage (72%), blastocyst from the initial number of cumulus oocyte complexes (46%) or from the cleaved ones (63%), hatching rate (69%), and the total number of blastomeres (187). In experiment 2, there was no difference of MII rate between slaughterhouse oocytes cultured for 18 or 22 hours, whereas the MII rate increased significantly (P < 0.05) between 18 and 22 hours for LOPU oocytes in the simplified medium. Moreover, slaughterhouse oocytes cultured in simplified medium matured significantly faster than LOPU oocytes at 18 and 22 hours (P < 0.05). In experiment 3, cleavage rate was significantly greater (P < 0.001) in all four groups of embryos produced by PA than IVF. Interestingly, PA reached similar rates for slaughterhouse oocytes cultured in both media, but improved (P < 0.05) the cleavage rate of LOPU oocytes. Slaughterhouse oocytes had acceptable cleavage rate after IVF (∼67%), whereas LOPU oocytes displayed a lower one (∼38%), in contrast to cleavage after PA. The percentage of blastocysts in relation to cleaved embryos was not affected by the origin of the oocytes (P > 0.05). Therefore, slaughterhouse oocytes developed a greater proportion of blastocysts than LOPU ones, expressed as the percentage of total cumulus oocyte complexes entering to IVM. Vitrified-thawed blastocysts presented similar survival and hatching rates between the oocyte origin, media, or method of activation. In conclusion, slaughterhouse and LOPU derived oocytes may have different IVM kinetics and require different IVM and IVF conditions. Although the IVM and IVF systems still need improvements to enhance embryo yield, the in vitro development step is able to generate good quality embryos from LOPU-derived oocytes. Copyright © 2014 Elsevier Inc. All rights reserved.

Genetic parameters for oocyte number and embryo production within a bovine ovum pick-up-in vitro production embryo-production program.

PubMed

Merton, J S; Ask, B; Onkundi, D C; Mullaart, E; Colenbrander, B; Nielen, M

2009-10-15

Genetic factors influencing the outcome of bovine ovum pick-up-in vitro production (OPU-IVP) and its relation to female fertility were investigated. For the first time, genetic parameters were estimated for the number of cumulus-oocyte complexes (Ncoc), quality of cumulus-oocyte complexes (Qcoc), number and proportion of cleaved embryos at Day 4 (Ncleav(D4), Pcleav(D4)), and number and proportion of total and transferable embryos at Day 7 of culture (Nemb(D7), Pemb(D7) and NTemb(D7), PTemb(D7), respectively). Data were recorded by CRV (formally Holland Genetics) from the OPU-IVP program from January 1995 to March 2006. Data were collected from 1508 Holstein female donors, both cows and pregnant virgin heifers, with a total of 18,702 OPU sessions. Data were analyzed with repeated-measure sire models with permanent environment effect using ASREML (Holstein Friesian). Estimates of heritability were 0.25 for Ncoc, 0.09 for Qcoc, 0.19 for Ncleav(D4), 0.21 for Nemb(D7), 0.16 for NTemb(D7), 0.07 for Pcleav(D4), 0.12 for Pemb(D7), and 0.10 for PTemb(D7). Genetic correlation between Ncoc and Qcoc was close to zero, whereas genetic correlations between Ncoc and the number of embryos were positive and moderate to high for Nemb(D7) (0.47), NTemb(D7) (0.52), and Ncleav(D4) (0.85). Genetic correlations between Ncoc and percentages of embryos (Pcleav(D4), Pemb(D7), and PTemb(D7)) were all close to zero. Phenotypic correlations were in line with genetic correlations. Genetic and phenotypic correlations between Qcoc and all other traits were not significant except for the phenotypic correlations between Qcoc and number of embryos, which were negative and low to moderate for Nemb(D7) (-0.20), NTemb(D7) (-0.24), and Ncleav(D4) (-0.43). Results suggest that cumulus-oocyte complex (COC) quality, based on cumulus investment, is independent from the total number of COCs collected via OPU and that in general, a higher number of COCs will lead to a higher number of embryos produced. The correlation between the estimated breeding values for Ncoc and PTemb(D7) of sires in this study and the sires breeding index for female-fertility based on the Dutch cattle population was close to zero. This study revealed OPU-IVP traits (Nemb(D7), NTemb(D7), and Ncoc) that could be of potential value for selection. Introduction of such traits in breeding programs would enhance the number of offspring from superior donors as well as improve the cost efficiency of OPU-IVP programs.

Parental occupation and childhood cancer: review of epidemiologic studies.

PubMed Central

Savitz, D A; Chen, J H

1990-01-01

Parental occupational exposures might affect childhood cancer in the offspring through genetic changes in the ovum or sperm or through transplacental carcinogenesis. The 24 published epidemiologic studies of this association have all used case-control designs, with controls generally selected from birth certificates or from general population sampling. Occupational exposures were inferred from job titles on birth certificates or through interviews. A large number of occupation-cancer associations have been reported, many of which were not addressed or not confirmed in other studies. Several associations have been found with consistency: paternal exposures in hydrocarbon-associated occupations, the petroleum and chemical industries, and especially paint exposures have been associated with brain cancer; paint exposures have also been linked to leukemias. Maternal exposures have received much less attention, but studies have yielded strongly suggestive results linking a variety of occupational exposures to leukemia and brain cancer. The primary limitations in this literature are the inaccuracy inherent in assigning exposure based on job title alone and imprecision due to limited study size. Although no etiologic associations have been firmly established by these studies, the public health concerns and suggestive data warrant continued research. PMID:2272330

Sperm and ova as property.

PubMed Central

Jansen, R P

1985-01-01

To whom do sperm and ova belong? Few tissues are produced by the human body with more waste than the germ cells. Yet dominion over the germ cells, and over the early embryo that results from their union in vitro, is behind much of the emotion that modern reproductive intervention can engender. The germ cells differ from other human tissues that can be donated or transplanted because they carry readily utilizable genetic information. Eventual expression of the germ cells' genetic potential is the legitimate concern and responsibility of their donors, although in the right circumstances the responsibility can by agreement be entrusted to institutions administering gamete or embryo donor programmes; these institutions, in turn, may need to assume responsibility for decisions if, in the case of embryo storage, the wishes of the two donors conflict. The fact of sperm and ovum ownership (and the genetic potential that goes with it) before individuals part with these tissues is beyond dispute. Some contentious issues may be clarified if this area of human dominion, namely control over genetic expression among offspring, is acknowledged to be the legitimate persisting concern of those who have produced sperm and ova after storage commences. PMID:3840532

Method for observing phase objects without halos and directional shadows

NASA Astrophysics Data System (ADS)

Suzuki, Yoshimasa; Kajitani, Kazuo; Ohde, Hisashi

2015-03-01

A new microscopy method for observing phase objects without halos and directional shadows is proposed. The key optical element is an annular aperture at the front focal plane of a condenser with a larger diameter than those used in standard phase contrast microscopy. The light flux passing through the annular aperture is changed by the specimen's surface profile and then passes through an objective and contributes to image formation. This paper presents essential conditions for realizing the method. In this paper, images of colonies formed by induced pluripotent stem (iPS) cells using this method are compared with the conventional phase contrast method and the bright-field method when the NA of the illumination is small to identify differences among these techniques. The outlines of the iPS cells are clearly visible with this method, whereas they are not clearly visible due to halos when using the phase contrast method or due to weak contrast when using the bright-field method. Other images using this method are also presented to demonstrate a capacity of this method: a mouse ovum and superimposition of several different images of mouse iPS cells.

Les électrolytes du sang et des sécrétions endométriales de la vache à la suite d'une glucocorticothérapie

PubMed Central

Ibrahim, M.; Guay, P.; Lamothe, P.

1972-01-01

The effect of two synthetic glucocorticoids, 9α-fluoroprednisolone acetate and dexamethasone (9α-fluoro, 16α-methylprednisolone) was studied on 12 normally breeding cows. Na, Mg, K, Ca and P concentrations were evaluated in the serum and in the uterine fluid at four stages of the estrous cycle. No significant changes were noted in Na and Ca concentration in either serum or uterine fluid. On the other hand, significant changes in the Mg, K and P were observed. Both drugs have caused a decrease in the content of Mg in the serum (P<0.05) during diestrus, a fall in the level of uterine K (P<0.01) during proestrus and a decrease of P during proestrus both in serum (P<0.001 after dexamethasone treatment) and in uterine fluid (P<0.001 for both drugs). It is obvious that the two drugs change the chemical composition of blood and endometrial secretions. This could jeopardize the survival of the spermatozoids and of the ovum as well as the fertilization and the implantation processes. PMID:4259930

[The early pregnancy factor (EPF) as an early marker of disorders in pregnancy].

PubMed

Straube, W; Römer, T; Zeenni, L; Loh, M

1995-01-01

The early pregnancy factor (EPF) seems to be very helpful in clinical applications such as early detection of pregnancy, differential diagnosis of failure of fertilization or implementation and prognosis of a fertilized ovum. Our purpose was to investigate the diagnostic value of single and serial measurement of EPF, especially in the differential diagnosis of abortion and extrauterine pregnancy. Women with a history of 6-16 weeks amenorrhoea with/without vaginal bleeding were included in the prospective study. The EPF-test system was carried out by means of the rosette inhibition method. EPF proved to be always positive in normal pregnant women and always negative in nonpregnant controls. In case of threatened abortion the prognosis was good, when the EPF values were positive, and poor when they became negative. Patients suffering from spontaneous and missed abortion mostly showed negative EPF-values. This was also true in ectopic pregnancies. The sensitivity and specificity of EPF-test system were 83%. The positive predictive value was observed to be 54% and the negative predictive value 95%. The EPF as an early embryonic signal may be a suitable parameter for the clinical use detecting pregnancy disturbances very early.

A longitudinal study of the control of renal and uterine hemodynamic changes of pregnancy.

PubMed

Ogueh, Onome; Clough, Angela; Hancock, Maggie; Johnson, Mark R

2011-01-01

We have determined the sequence and extent of maternal renal and uterine adaptation to pregnancy and examined the role of hormonal factors in their regulation. Renal bipolar diameter (RBD), renal artery resistance index (RARI), uterine artery pulsatility index (UAPI), and plasma relaxin, human chorionic gonadotropin (hCG), progesterone, estradiol, urea, and creatinine were measured longitudinally in women with normal spontaneous singleton pregnancies, in vitro fertilization (IVF) singleton pregnancies, ovum donation (OD) singleton pregnancies, and multiple pregnancies from prepregnancy to postpartum. There was a progressive increase in the RBD and the RARI and a decrease in the UAPI during pregnancy. These changes reversed toward prepregnancy levels by 6 weeks post delivery. There was no difference in the rate of change of RBD, RARI, and UAPI between spontaneous singleton, IVF singleton, OD singleton, and multiple pregnancies (p < 0.05), but relaxin was directly correlated to the RARI (r = 0.654, p = 0.015), and progesterone was inversely correlated to uterine artery PI (r = 0.554, p = 0.049). These data show that renal size and resistance to blood flow increase with advancing gestation, whereas the uterine artery resistance declined with gestation. These changes may be influenced by relaxin and progesterone.

Manipulation of follicle development to ensure optimal oocyte quality and conception rates in cattle.

PubMed

Baruselli, P S; Sá Filho, M F; Ferreira, R M; Sales, J N S; Gimenes, L U; Vieira, L M; Mendanha, M F; Bó, G A

2012-08-01

Over the last several decades, a number of therapies have been developed that manipulate ovarian follicle growth to improve oocyte quality and conception rates in cattle. Various strategies have been proposed to improve the responses to reproductive biotechnologies following timed artificial insemination (TAI), superovulation (SOV) or ovum pickup (OPU) programmes. During TAI protocols, final follicular growth and size of the ovulatory follicle are key factors that may significantly influence oocyte quality, ovulation, the uterine environment and consequently pregnancy outcomes. Progesterone concentrations during SOV protocols influence follicular growth, oocyte quality and embryo quality; therefore, several adjustments to SOV protocols have been proposed depending on the animal category and breed. In addition, the success of in vitro embryo production is directly related to the number and quality of cumulus oocyte complexes harvested by OPU. Control of follicle development has a significant impact on the OPU outcome. This article discusses a number of key points related to the manipulation of ovarian follicular growth to maximize oocyte quality and improve conception rates following TAI and embryo transfer of in vivo- and in vitro-derived embryos in cattle. © 2012 Blackwell Verlag GmbH.

Is there a ‘new ethics of abortion'?

PubMed Central

Gillon, R.

2001-01-01

This paper argues that the central issue in the abortion debate has not changed since 1967 when the English parliament enacted the Abortion Act. That central issue concerns the moral status of the human fetus. The debate here is not, it is argued, primarily a moral debate, but rather a metaphysical debate and/or a theological debate—though one with massive moral implications. It concerns the nature and attributes that an entity requires to have "full moral standing" or "moral inviolability" including a "right to life". It concerns the question when, in its development from newly fertilised ovum to unequivocally mature, autonomous morally inviolable person does a human being acquire that nature and those attributes, and thus a "right to life". The paper briefly reviews standard answers to these questions, outlining some problems associated with each. Finally there is a brief discussion of one way in which the abortion debate has changed since 1967—notably in the increasingly vociferous claim, especially from disability rights sectors, that abortion on grounds of fetal abnormality implies contempt for and rejection of disabled people—a claim that is rebutted. Key Words: Abortion • moral status of fetus • abortion of abnormal fetuses PMID:11574651

Pregnancy Outcome Using General Anesthesia Versus Spinal Anesthesia for In Vitro Fertilization

PubMed Central

Azmude, Azra; Agha'amou, Shahrzad; Yousefshahi, Fardin; Berjis, Katayoun; Mirmohammad'khani, Majid; Sadaat'ahmadi, Farahnaz; Ghods, Kamran; Dabbagh, Ali

2013-01-01

Background There is a considerable rate of fertility failure and this causes a great burden of untoward effects for patients. Usually a considerable number of these patients undergo anesthesia for their treatment. Objectives This study was designed to compare the effects of general and spinal anesthesia on these patients. Patients and Methods In a randomized clinical trial, after taking informed written consent from the patients, 200 patients entered the study; 100 in each. During a 2 year period, women aged 20 to 40 years entered the study (one group receiving spinal anesthesia and the other, receiving general anesthesia). Ovum retrieval protocols were the same. Nonparametric and parametric analyses were used for data analysis. P value less than 0.05 was considered significant. Results There was no difference between the two groups regarding demographic variables. 15 of 100 patients (15%) in the general anesthesia group and 27 of 100 patients (27%) in the spinal anesthesia group had successful pregnancy after IVF; so, spinal anesthesia increased significantly the chance of IVF success (P value < 0.001; Chi Square). Conclusions The results of this study demonstrated that spinal anesthesia increased the chance of fertilization success. PMID:24282775

Use of FSH in two different regimens for ovarian superstimulation prior to ovum pick up and in vitro embryo production in Holstein cows.

PubMed

da Silva, Júlio César Barboza; Ferreira, Roberta Machado; Maturana Filho, Milton; Naves, Julianne de Rezende; Santin, Thiago; Pugliesi, Guilherme; Madureira, Ed Hoffmann

2017-03-01

We aimed with the present study to evaluate the effects of FSH treatment (200 mg) split in four or six administrations on ovarian follicle stimulation and in vitro oocyte competence for embryo production in dairy cows with synchronized follicular wave emergence. On random days of the estrous cycle (Day 0), non-lactating Holstein cows received a progesterone (P4)-releasing intravaginal device and 2 mg estradiol benzoate IM. On Day 3, they received 0.530 mg sodium cloprostenol (PGF2α) IM. Control cows (n = 35) received no further treatments, whereas FSH-treated cows received 200 mg FSH split in four (FSH4 group; n = 33) or six (FSH6 group; n = 33) administration regimens. Starting on Day 4, cows in FSH4 group received 200 mg FSH split in four equivalent doses of 50 mg 12 h apart. Cows in FSH6 group received the same total FSH dose split in six equivalent doses of 33.3 mg 12 h apart, but treatments started on Day 3. On Day 7 AM (36 h of "coasting" period for FSH-treated groups), the P4 devices were removed and cows were subjected to ovum pick up (OPU). Viable oocytes were in vitro fertilized using sexed-sorted semen. Although FSH treatment did not (P > 0.1) increase the total number of follicles (Control, 53.2 ± 4.5 vs. FSH-treated, 51.4 ± 3.1), the two hormonal stimulation regimens, FSH4 and FSH6, increased the number of medium follicles (6-10 mm; 5.2 ± 0.5 vs. 18.1 ± 1.4; P < 0.0001) and reduced the number of small follicles (2-5.9 mm; 46.3 ± 5.1 vs. 31.0 ± 2.4 P < 0.0001). Also, FSH treatment or regimen did not increase (P > 0.1) the number of viable oocytes (Control, 12.6 ± 1.26 vs. FSH-treated, 12.70 ± 1.03), recovery rate (Control, 36.5% vs. FSH-treated, 36%) and the number of in vitro produced blastocyst (Control, 4.1 ± 0.52 vs. FSH-treated 4.3 ± 0.5). We concluded that FSH stimulation protocol proposed herein is effective to stimulate the growth of small antral follicle population prior to OPU, but it was ineffective to improve in vitro oocyte competence for embryo production in non-lactating Holstein cows with synchronized follicular wave emergence. Copyright © 2017 Elsevier Inc. All rights reserved.

Efficacy of a single intramuscular injection of porcine FSH in hyaluronan prior to ovum pick-up in Holstein cattle.

PubMed

Vieira, L M; Rodrigues, C A; Castro Netto, A; Guerreiro, B M; Silveira, C R A; Freitas, B G; Bragança, L G M; Marques, K N G; Sá Filho, M F; Bó, G A; Mapletoft, R J; Baruselli, P S

2016-03-15

Plasma FSH profiles, in vitro embryo production (IVP) after ovum pickup (OPU), and establishment of pregnancy with IVP embryos were compared in untreated Holstein oocyte donors and those superstimulated with multiple injections or a single intramuscular (IM) injection of porcine FSH (pFSH) in hyaluronan (HA). Plasma FSH profiles were determined in 23 heifers randomly allocated to one of four groups. Controls received no treatment, whereas the F200 group received 200 mg of pFSH in four doses, 12 hours apart. The F200HA and F300HA groups received 200- or 300-mg pFSH in 5 mL or 7.5 mL, respectively of a 0.5% HA solution by a single IM injection. Plasma FSH levels were determined before the first pFSH treatment and every 6 hours over 96 hours. All data were analyzed by orthogonal contrasts. Circulating FSH area under curve (AUC) in pFSH-treated animals was greater than that in the control group (P = 0.02). Although the AUC did not differ among FSH-treated groups (P = 0.56), the total period with elevated plasma FSH was greater in the F200 group than in the HA groups (P < 0.0001). However, the F300HA group had a greater AUC than the F200HA group (P = 0.006), with a similar total period with elevated plasma FSH (P = 0.17). The IVP was performed in 90 nonlactating Holstein cows randomly allocated to one of the four treatment groups as in the first experiment. A greater proportion of medium-sized (6-10 mm) follicles was observed in cows receiving pFSH, regardless of the treatment group (P < 0.0001). Also, numbers of follicles (P = 0.01), cumulus-oocyte complexes (COCs) retrieved (P = 0.01) and matured (P = 0.02), cleavage rates (P = 0.002), and blastocysts produced per OPU session (P = 0.06) were greater in cows receiving pFSH, regardless of the treatment group. Cows in the F200HA group had a greater recovery rate (P = 0.009), number of COCs cultured (P = 0.04), and blastocysts produced per OPU session (P = 0.06) than cows in the F300HA group. Similar pregnancy rates were observed 50 to 60 days after transferring IVP embryos from donors in the different treatment groups (P > 0.05). In conclusion, a single IM injection of pFSH combined in 0.5% HA resulted in similar plasma FSH profiles as twice-daily pFSH treatments. Treatment of nonlactating donors with pFSH, with or without HA, resulted in increased IVP over untreated controls. A single dose of 200 mg of pFSH in 0.5% HA resulted in greater IVP than 300-mg pFSH in HA. Finally, pregnancy rates with IVP embryos were similar, regardless donor treatment. Copyright © 2016 Elsevier Inc. All rights reserved.

[Identification of NMDA receptor in normal bovine ovary and ovum].

PubMed

Tachibana, Naoko; Ikeda, Shu-ichi

2014-01-01

To clarify the pathogenesis of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis in patients without ovarian teratoma, we investigate normal human ovary, normal bovine ovary and bovine ova. On the basis of immunohistochemical studies, normal human ovary expressed NR2B epitope in primordial oocytes. The results of SDS-PAGE and immunoblotting using bovine ovarian tissues and ova, we identified two bands of NR1 and NR2B. Moreover, reverse phase liquid chromatography coupled to tandem mass spectrometry showed peptides fractions of NR1, NR2A, NR2B and NR2C. Immunocytochemical study disclosed that normal bovine oocyte has a strong affinity for a patient's disease-specific IgG. Anti-NMDAR encephalitis involves mainly young women who are in their reproductive age. Ovarian teratoma is important as simultaneous tumor, the percentage of patients with ovarian teratoma is less than 40%. It is obvious that the origin of ovarian teratoma is oocyte. So the existence of NMDAR in normal oocytes is very important to assert that ovary itself is the antigen presenting tissue. And also it is helpful to explain why young women are mainly affected from this disease. It seems to conclude that anti-NMDAR encephalitis is one form of autoimmune synaptic encephalitis and that the antigen presenting tissue is ovary itself.

Therapeutic abortion in Islam: contemporary views of Muslim Shiite scholars and effect of recent Iranian legislation

PubMed Central

Hedayat, K M; Shooshtarizadeh, P; Raza, M

2006-01-01

Abortion is forbidden under normal circumstances by nearly all the major world religions. Traditionally, abortion was not deemed permissible by Muslim scholars. Shiite scholars considered it forbidden after implantation of the fertilised ovum. However, Sunni scholars have held various opinions on the matter, but all agreed that after 4 months gestation abortion was not permitted. In addition, classical Islamic scholarship had only considered threats to maternal health as a reason for therapeutic abortion. Recently, scholars have begun to consider the effect of severe fetal deformities on the mother, the families and society. This has led some scholars to reconsider the prohibition on abortion in limited circumstances. This article reviews the Islamic basis for the prohibition of abortion and the reasons for its justification. Contemporary rulings from leading Shiite scholars and from the Sunni school of thought are presented and reviewed. The status of abortion in Muslim countries is reviewed, with special emphasis on the therapeutic abortion law passed by the Iranian Parliament in 2003. This law approved therapeutic abortion before 16 weeks of gestation under limited circumstances, including medical conditions related to fetal and maternal health. Recent measures in Iran provide an opportunity for the Muslim scholars in other countries to review their traditional stance on abortion. PMID:17074823

Therapeutic abortion in Islam: contemporary views of Muslim Shiite scholars and effect of recent Iranian legislation.

PubMed

Hedayat, K M; Shooshtarizadeh, P; Raza, M

2006-11-01

Abortion is forbidden under normal circumstances by nearly all the major world religions. Traditionally, abortion was not deemed permissible by Muslim scholars. Shiite scholars considered it forbidden after implantation of the fertilised ovum. However, Sunni scholars have held various opinions on the matter, but all agreed that after 4 months gestation abortion was not permitted. In addition, classical Islamic scholarship had only considered threats to maternal health as a reason for therapeutic abortion. Recently, scholars have begun to consider the effect of severe fetal deformities on the mother, the families and society. This has led some scholars to reconsider the prohibition on abortion in limited circumstances. This article reviews the Islamic basis for the prohibition of abortion and the reasons for its justification. Contemporary rulings from leading Shiite scholars and from the Sunni school of thought are presented and reviewed. The status of abortion in Muslim countries is reviewed, with special emphasis on the therapeutic abortion law passed by the Iranian Parliament in 2003. This law approved therapeutic abortion before 16 weeks of gestation under limited circumstances, including medical conditions related to fetal and maternal health. Recent measures in Iran provide an opportunity for the Muslim scholars in other countries to review their traditional stance on abortion.

Modeling man: the monkey colony at the Carnegie Institution of Washington's Department of Embryology, 1925-1971.

PubMed

Wilson, Emily K

2012-01-01

Though better recognized for its immediate endeavors in human embryo research, the Carnegie Department of Embryology also employed a breeding colony of rhesus macaques for the purposes of studying human reproduction. This essay follows the course of the first enterprise in maintaining a primate colony for laboratory research and the overlapping scientific, social, and political circumstances that tolerated and cultivated the colony's continued operation from 1925 until 1971. Despite a new-found priority for reproductive sciences in the United States, by the early 1920s an unfertilized human ovum had not yet been seen and even the timing of ovulation remained unresolved. Progress would require an organized research approach that could extend beyond the limitations of working with scant and inherently restrictive human subjects or with common lab mammals like mice. In response, the Department of Embryology, under the Carnegie Institution of Washington (CIW), instituted a novel methodology using a particular primate species as a surrogate in studying normal human reproductive physiology. Over more than 40 years the monkey colony followed an unpremeditated trajectory that would contribute fundamentally to discoveries in human reproduction, early embryo development, reliable birth control methods, and to the establishment of the rhesus macaque as a common model organism.

The Origin and Fate of Annulate Lamellae in Maturing Sand Dollar Eggs

PubMed Central

Merriam, R. W.

1959-01-01

Electron micrograph evidence is presented that the nuclear envelope of the mature ovum of Dendraster excentricus is implicated in a proliferation of what appear as nuclear envelope replicas in the cytoplasm. The proliferation is associated with intranuclear vesicles which apparently coalesce to form comparatively simple replicas of the nuclear envelope closely applied to the inside of the nuclear envelope. The envelope itself may become disorganized at the time when fully formed annulate lamellae appear on the cytoplasmic side and parallel with it. The concept of interconvertibility of general cytoplasmic vesicles with most of the membrane systems of the cytoplasm is presented. The structure of the annuli in the annulate lamellae is shown to include small spheres or vesicles of variable size embedded in a dense matrix. Dense particles which are about 150 A in diameter are often found closely associated with annulate lamellae in the cytoplasm. Similar structures in other echinoderm eggs are basophilic. In this species, unlike other published examples, the association apparently takes place in the cytoplasm only after the lamellae have separated from the nucleus. If 150 A particles are synthesized by annulate lamellae, as their close physical relationship suggests, then in this species at least the necessary synthetic mechanisms and specificity must reside in the structure of annulate lamellae. PMID:13630942

Sentan: A Novel Specific Component of the Apical Structure of Vertebrate Motile Cilia

PubMed Central

Yuba-Kubo, Akiko; Tsukita, Sachiko; Tsukita, Shoichiro; Amagai, Masayuki

2008-01-01

Human respiratory and oviductal cilia have specific apical structures characterized by a narrowed distal portion and a ciliary crown. These structures are conserved among vertebrates that have air respiration systems; however, the molecular components of these structures have not been defined, and their functions are unknown. To identify the molecular component(s) of the cilia apical structure, we screened EST libraries to identify gene(s) that are exclusively expressed in ciliated tissues, are transcriptionally up-regulated during in vitro ciliogenesis, and are not expressed in testis (because sperm flagella have no such apical structures). One of the identified gene products, named sentan, was localized to the distal tip region of motile cilia. Using anti-sentan polyclonal antibodies and electron microscopy, sentan was shown to localize exclusively to the bridging structure between the cell membrane and peripheral singlet microtubules, which specifically exists in the narrowed distal portion of cilia. Exogenously expressed sentan showed affinity for the membrane protrusions, and a protein–lipid binding assay revealed that sentan bound to phosphatidylserine. These findings suggest that sentan is the first molecular component of the ciliary tip to bridge the cell membrane and peripheral singlet microtubules, making the distal portion of the cilia narrow and stiff to allow for better airway clearance or ovum transport. PMID:18829862

Combined ovulation triggering with GnRH agonist and hCG in IVF patients.

PubMed

Kasum, Miro; Kurdija, Kristijan; Orešković, Slavko; Čehić, Ermin; Pavičić-Baldani, Dinka; Škrgatić, Lana

2016-11-01

The aim of the review is to analyse the combination of a gonadotrophin releasing hormone (GnRH) agonist with a human chorionic gonadotrophin (hCG) trigger, for final oocyte maturation in in vitro fertilisation (IVF) cycles. The concept being a ''dual trigger'' combines a single dose of the GnRH agonist with a reduced or standard dosage of hCG at the time of triggering. The use of a GnRH agonist with a reduced dose of hCG in high responders demonstrated luteal phase support with improved pregnancy rates, similar to those after conventional hCG and a low risk of ovarian hyperstimulation syndrome (OHSS). The administration of a GnRH agonist and a standard hCG in normal responders, demonstrated significantly improved live-birth rates and a higher number of embryos of excellent quality, or cryopreserved embryos. The concept of the ''double trigger" represents a combination of a GnRH agonist and a standard hCG, when used 40 and 34 h prior to ovum pick-up, respectively. The use of the ''double trigger" has been successfully offered in the treatment of empty follicle syndrome and in patients with a history of immature oocytes retrieved or with low/poor oocytes yield. Further prospective studies are required to confirm the aforementioned observations prior to clinical implementation.

A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation

PubMed Central

Roldan, Eduardo R. S.

2016-01-01

Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in non-capacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage. PMID:26936246

Determining the need for rescue intracytoplasmic sperm injection in partial fertilisation failure during a conventional IVF cycle.

PubMed

Cao, S; Wu, X; Zhao, C; Zhou, L; Zhang, J; Ling, X

2016-12-01

To explore the need for rescue intracytoplasmic sperm injection (ICSI) in cases of partial fertilisation failure during a conventional in vitro fertilisation cycle, rescue ICSI was performed for cycles with a fertilisation rate of <50%. The data were divided into three groups based on the fertilisation rate: group 1 (0%), group 2 (<25%) and group 3 (>25%). The impact of rescue ICSI on each group was then analysed in terms of ovum fertilisation, embryo development, embryo utilisation and selection of embryos for transfer. Rescue ICSI was performed on 1831 unfertilised oocytes from 313 cycles. The fertilisation rates for group 1, group 2 and group 3 were 74.66, 68.35 and 65.46%, and the rate of polyploidy in the three groups was 8.55, 11.33, and 14.47%. The percentage of embryos that can be transferred from rescue ICSI for group 2 was 38.33%, and this value was higher than those of the other two groups. It is concluded that rescue ICSI is not recommended for patients with an IVF rate of >25% as the procedure is associated with a greater risk and low returns. However, it is feasible to perform rescue ICSI for patients with IVF rates of <25%. © 2016 Blackwell Verlag GmbH.

Levonorgestrel decreases cilia beat frequency of human fallopian tubes and rat oviducts without changing morphological structure.

PubMed

Zhao, Weihong; Zhu, Qian; Yan, Mingxing; Li, Cheng; Yuan, Jiangjing; Qin, Guojuan; Zhang, Jian

2015-02-01

Levonorgestrel, a derivative of progesterone, effectively protects women against unwanted pregnancy as an emergency contraceptive. Previous studies have not been successful in determining the mechanism by which levonorgestrel acts. In the present study we analysed cilia beat action and cilia morphology following levonorgestrel exposure in vitro and in vivo using both light and electron microscopy. There was a significant decrease in the ciliary beat frequency (CBF) of human fallopian tubes between mucosal explants bathed in 5 μmol/L levonorgestrel and those bathed in medium alone (P < 0.05). There was a tendency for CBF to decrease more in the ampulla than in isthmus, but there were no differences between the proliferative and secretory phases. In rat oviducts, levonorgestrel produced a similar reduction in CBF (~ 10%) compared with the saline control group (P < 0.05). Histological and ultrastructural analysis demonstrated no changes in the percentage of ciliated cells or in the classic '9 + 2' structure of cilia following levonorgestrel treatment in either system. Thus, levonorgestrel reduces CBF without damaging cilia morphology. Decreases in CBF may indicate a pathological role for levonorgestrel in the transportation of the ovum and zygote in the fallopian tube. © 2014 Wiley Publishing Asia Pty Ltd.

Genetics and biology of human ovarian teratomas. I. Cytogenetic analysis and mechanism of origin.

PubMed Central

Surti, U; Hoffner, L; Chakravarti, A; Ferrell, R E

1990-01-01

One hundred and two benign, mature ovarian teratomas and two immature, malignant teratomas were karyotyped and scored for centromeric heteromorphisms as part of an ongoing project to determine the chromosomal karyotype and the genetic origin of ovarian teratomas and to assess their utility for gene-centromere mapping. Karyotypic analysis of the benign cases revealed 95 46,XX teratomas and 7 chromosomally abnormal teratomas (47,XXX, 47,XX,+8 [two cases], 47,XX,+15, 48,XX,+7,+12 91,XXXX,-13 [mosaic], 47,XX,-15,+21,+mar). Our study reports on the first cases of tetraploidy and structural rearrangement in benign ovarian teratomas. The two immature cases had modal chromosome numbers of 78 and 49. Centromeric heteromorphisms that were heterozygous in the host were homozygous in 65.2% (n = 58) of the benign teratomas and heterozygous in the remaining 34.8% (n = 31). Chromosome 13 heteromorphisms were the most informative, with 72.7% heterozygosity in hosts. The cytogenetic data indicate that 65% of teratomas are derived from a single germ cell after meiosis I and failure of meiosis II (type II) or endoreduplication of a mature ovum (type III); 35% arise by failure of meiosis I (type I) or mitotic division of premeiotic germ cells (type IV). Images Figure 1 PMID:2220805

Peripheral Reproductive Organ Health and Melatonin: Ready for Prime Time

PubMed Central

Reiter, Russel J.; Rosales-Corral, Sergio A.; Manchester, Lucien C.; Tan, Dun-Xian

2013-01-01

Melatonin has a wide variety of beneficial actions at the level of the gonads and their adnexa. Some actions are mediated via its classic membrane melatonin receptors while others seem to be receptor-independent. This review summarizes many of the published reports which confirm that melatonin, which is produced in the ovary, aids in advancing follicular maturation and preserving the integrity of the ovum prior to and at the time of ovulation. Likewise, when ova are collected for in vitro fertilization-embryo transfer, treating them with melatonin improves implantation and pregnancy rates. Melatonin synthesis as well as its receptors have also been identified in the placenta. In this organ, melatonin seems to be of particular importance for the maintenance of the optimal turnover of cells in the villous trophoblast via its ability to regulate apoptosis. For male gametes, melatonin has also proven useful in protecting them from oxidative damage and preserving their viability. Incubation of ejaculated animal sperm improves their motility and prolongs their viability. For human sperm as well, melatonin is also a valuable agent for protecting them from free radical damage. In general, the direct actions of melatonin on the gonads and adnexa of mammals indicate it is an important agent for maintaining optimal reproductive physiology. PMID:23549263

A case report of a patient with high β-hCG levels after operation because of primary broad ligament pregnancy.

PubMed

Zu, M; Zhao, G Q; Liu, Z Q; Zhang, H T; Chen, L; Zhao, D H

2017-01-01

A broad ligament pregnancy is an extremely rare condition and diagnosis is frequently missed and finally made during laparotomy. This is a case of a young patient with high serum beta-human chorionic gonadotropin (β-hCG) levels after operation because of broad ligament pregnancy. A 31-year-old multipara complained of intermittent lower abdominal pain with vaginal bleeding for four months. A color ultrasonography revealed a cystic mass in the left attachment area, indicating an interstitial tubal pregnancy. However, trophoblastic disease could not be excluded. She accepted conservative treatment with methotrexate (MTX) at first, but observation showed that conservative treatment was slow and accompanied with liver function damage. Therefore, exploratory laparotomy was performed. Intraoperative situations and postoperative pathology confirmed broad ligament pregnancy. Her serum p- hCG was sustained at a high level for three months after operation. Her examinations of serum, CT, and ultrasonography could explain this situation. Primary broad ligament pregnancy refers to pregnancy where implantation of the fertilized ovum occurs directly between the two leaves of the broad ligament. The gravid substance was removed, however serum β-hCG could not gradually re- turn to normal levels. This case should be followed-up closely to prevent adverse outcomes.

In vitro production of small ruminant embryos: late improvements and further research.

PubMed

de Souza-Fabjan, Joanna Maria Gonçalves; Panneau, Barbara; Duffard, Nicolas; Locatelli, Yann; de Figueiredo, José Ricardo; Freitas, Vicente José de Figueirêdo; Mermillod, Pascal

2014-06-01

Beyond the potential use of in vitro production of embryos (IVP) in breeding schemes, embryos are also required for the establishment of new biotechnologies such as cloning and transgenesis. Additionally, the knowledge of oocyte and embryo physiology acquired through IVP techniques may stimulate the further development of other techniques such as marker assisted and genomic selection of preimplantation embryos, and also benefit assisted procreation in human beings. Efficient in vitro embryo production is currently a major objective for livestock industries, including small ruminants. The heterogeneity of oocytes collected from growing follicles by laparoscopic ovum pick up or in ovaries of slaughtered females, remains an enormous challenge for IVM success, and still limits the rate of embryo development. In addition, the lower quality of the IVP embryos, compared with their in vivo-derived counterparts, translates into poor cryosurvival, which restricts the wider use of this promising technology. Therefore, many studies have been reported in an attempt to determine the most suitable conditions for IVM, IVF, and in vitro development to maximize embryo production rate and quality. This review aims to present the current panorama of IVP production in small ruminants, describing important steps for its success, reporting the recent advances and also the main obstacles identified for its improvement and dissemination. Copyright © 2014 Elsevier Inc. All rights reserved.

Ultrasonographic-guided retrieval of cumulus oocyte complexes after super-stimulation in dromedary camel (Camelus dromedarius).

PubMed

Wani, N A; Skidmore, J A

2010-08-01

In Experiment 1, studies were conducted to apply the transvaginal ultrasound guided ovum pick-up (OPU) technique in dromedary camels after their ovarian super-stimulation and in vivo oocyte maturation. In Experiment 2, the developmental potential of two commonly used oocyte types, i.e., in vivo matured oocytes collected by OPU and abattoir derived in vitro-matured oocytes was compared after their chemical activation. In Experiment 3, developmental competence of oocytes collected from super-stimulated camels by OPU, matured either in vivo or in vitro, was compared after their chemical activation. Mature female dromedary camels super-stimulated with a combination of eCG and pFSH were given an injection of 20 microg of the GnRH analogue, buserelin 24, 26, or 28 h before the scheduled OPU. For collection of cumulus oocyte complexes (COCs) the transducer was guided through the vulva into the cranial most portion of the vagina and 17-gauge, 55 cm single-lumen needle was placed in the needle guide of the ultrasound probe and advanced through the vaginal fornix and into the follicle. Follicular fluid was aspirated using a regulated vacuum pump into tubes containing embryo-flushing media. Aspirates were searched for COCs using a stereomicroscope, and they were then denuded of cumulus cells by hyaluronidase and repeated pipetting. The oocytes were classified as mature (with a visible polar body), immature (with no visible polar body), activated (with divided or fragmented ooplasm) and others (degenerated and abnormal). Overall an average of 12.12 +/- 7.9 COCs were aspirated per animal with an oocyte recovery rate from the aspirated follicles of about 77%. The majority (> 90%) of the collected COCs by OPU were with loose and expanded cumulus cells. The proportion of matured oocytes obtained at 28-29 h (91.2 +/- 4.1) and 26-27 h (82.1 +/- 3.4) were higher (P < 0.005) when compared with those obtained at 24-25 h (40.4 +/- 16.3) after GnRH administration. In Experiment 2, a higher proportion (P < 0.05) of in vivo matured oocytes cleaved (84.6 +/- 2.1 vs. 60.9 +/- 6.6) and developed to blastocyst stages (52.4 +/- 4.1 vs. 30.5 +/- 3.3) when compared with in vitro matured oocytes collected from slaughterhouse ovaries. In Experiment 3, no difference was observed between the developmental competences of oocytes, collected from super stimulated camels, matured in vitro with those collected after their in vivo maturation. In conclusion, about 80-90% mature oocytes can be collected by ultrasound guided transvaginal ovum pick-up from super-stimulated dromedary camels 26-28 h after GnRH administration. The developmental response, to chemical activation, of in vivo matured oocytes collected by ultrasound guided transvaginal OPU is better than in vitro matured oocytes obtained from slaughterhouse ovaries. However, no difference was observed in the developmental competence of oocytes collected by OPU whether they were matured in vivo or in vitro. Copyright 2010 Elsevier Inc. All rights reserved.

Formation and regression of the corpus luteum of the American alligator

USGS Publications Warehouse

Guillette, L.J.; Woodward, A.R.; You-Xiang, Q.; Cox, M.C.; Matter, J.H.; Gross, T.S.

1995-01-01

Luteal morphology of the American alligator is unique when compared to other reptiles but is similar to that of its phylogenetic relatives, the birds. The theca is extensively hypertrophied, but the granulosa never fills the cavity formed following the ovulation of the ovum. The formation of the corpus luteum (CL) is correlated with elevated plasma progesterone concentrations, which decline dramatically after oviposition with the onset of luteolysis. Unlike those of most other reptiles, the central luteal cell mass is composed of two cell types; one presumably is derived from the granulosa, whereas the other is from the theca interna. Both cell types are present throughout gravidity but only one cell type is seen during mid to late luteolysis. A significant decline in luteal volume occurs following oviposition and continues throughout the post-oviposition period. The fastest decline in luteal volume occurs in the month immediately after oviposition; this rate then slows. Luteolysis appears to continue for a year or more following oviposition, as distinct structures of luteal origin can still be identified in animals 9 months after oviposition. The size of persistent CL can be used to determine whether a given female oviposited during the previous nesting season. Females with CL having volumes greater than 0.2 cm2 or CL diameters greater than 0.4 cm were active the previous season. 

In vitro fertilization experiments using sockeye salmon reveal that bigger eggs are more fertilizable under sperm limitation

PubMed Central

Macfarlane, Christopher P.; Hoysak, Drew J.; Liley, N. Robin; Gage, Matthew J.G.

2009-01-01

Although theory and widespread evidence show that the evolution of egg size is driven primarily by offspring and maternal fitness demands, an additional explanation invokes sperm limitation as a selective force that could also influence egg size optima. Levitan proposed that constraints from gamete encounter in external fertilization environments could select for enlargement of ova to increase the physical size of the fertilization target. We test this theory using in vitro fertilization experiments in an externally fertilizing fish. Sockeye salmon (Onchorhyncus nerka) females show considerable between-individual variation in ovum size, and we explored the consequences of this natural variation for the fertilization success of individual eggs under conditions of sperm limitation. By engineering consistent conditions where in vitro fertilization rate was always intermediate, we were able to compare the sizes of fertilized and unfertilized eggs across 20 fertilization replicates. After controlling for any changes in volume through incubation, results showed that successfully fertilized eggs were significantly larger than the eggs that failed to achieve fertilization. Under conditions without sperm limitation, fertility was unaffected by egg size. Our findings therefore support Levitan's theory, demonstrating empirically that some element of egg size variation could be selected by fertilization demands under sperm limitation. However, further research on sperm limitation in natural spawnings is required to assess the selective importance of these results. PMID:19364734

In-vitro maturation of germinal vesicle and metaphase I eggs prior to cryopreservation optimizes reproductive potential in patients undergoing fertility preservation.

PubMed

Lee, Joseph A; Sekhon, Lucky; Grunfeld, Lawrence; Copperman, Alan B

2014-06-01

To evaluate current and previous findings related to a timely implementation of in-vitro maturation (IVM) of germinal vesicle, metaphase I and metaphase II oocytes with an optimal cryopreservation to determine whether IVM should be attempted prior to (fresh IVM) or IVM after cryopreservation (postthaw IVM). Mitochondrion, chromatin and spindle formation in both groups were interpreted from referenced studies to establish best management of all oocytes. The postthaw survival of germinal vesicle, metaphase I, fresh IVM-metaphase II and control metaphase II oocytes did not differ significantly [83.3% (n=9), 86.7% (n=12), 83% (n=57) and 86% (n=68), respectively]. Overall, combined survival and maturation were significantly higher (P<0.05) in the fresh IVM group at 63.8% (44 of 69) compared with the postthaw IVM group at 33.3% (nine of 27). Conservation of retrieved immature oocytes after vaginal oocyte retrieval has become a major concern for patients, as they strive to maximize the reproductive viability of all oocytes obtained during treatment. Oocyte cryopreservation is important for patients at risk of ovarian cancer, elective fertility preservation and potentially for ovum donation. The superior maturation rate of germinal vesicle and metaphase I oocytes in the fresh IVM vs. postthaw groups provides strong impetus to mature oocytes to the metaphase II stage prior to cryopreservation.

Re-evaluation of post-wash sperm is a helpful tool in the decision to perform in vitro fertilisation or intracytoplasmic sperm injection.

PubMed

Wiser, A; Ghetler, Y; Gonen, O; Piura, E; Berkovits, A; Itskovich, A; Rom, T; Shulman, A

2012-04-01

The aim of this study was to find discriminatory parameters, based on sperm characteristics on the day of ovum pickup, that can help guide the decision to perform either intracytoplasmic sperm injection (ICSI) or in vitro fertilisation (IVF). We evaluated 112 cycles fertilised with both regular and ICSI insemination during the same cycle. A total of 112 cycles were analysed. In 62 cycles, fertilisation was obtained with both ICSI and IVF, and in 50 cycles, fertilisation was obtained by ICSI alone. The sperm samples were re-evaluated after the preparation process. The mean initial total motile sperm count (TMSC) was 66.3 × 10(6) ± 47.5 in the group that underwent both methods and 23.1 × 10(6) ± 20.4 in the ICSI only group (P < 0.05). After sperm preparation, the mean post-wash TMSC was 4.4 × 10(6) ± 3.4 and 1.06 × 10(6) ± 0.9 respectively (P < 0.05). A cutoff of 1.5 × 10(6) or fewer sperm after preparation as an indicator for ICSI has a sensitivity of 80% and a specificity of 77%. Re-evaluation of TMSC can prevent unexpected fertilisation failure. Fewer than 1.5 million TMSC after wash should be considered an indication for ICSI fertilisation. © 2011 Blackwell Verlag GmbH.

In vitro fertilization experiments using sockeye salmon reveal that bigger eggs are more fertilizable under sperm limitation.

PubMed

Macfarlane, Christopher P; Hoysak, Drew J; Liley, N Robin; Gage, Matthew J G

2009-07-07

Although theory and widespread evidence show that the evolution of egg size is driven primarily by offspring and maternal fitness demands, an additional explanation invokes sperm limitation as a selective force that could also influence egg size optima. Levitan proposed that constraints from gamete encounter in external fertilization environments could select for enlargement of ova to increase the physical size of the fertilization target. We test this theory using in vitro fertilization experiments in an externally fertilizing fish. Sockeye salmon (Onchorhyncus nerka) females show considerable between-individual variation in ovum size, and we explored the consequences of this natural variation for the fertilization success of individual eggs under conditions of sperm limitation. By engineering consistent conditions where in vitro fertilization rate was always intermediate, we were able to compare the sizes of fertilized and unfertilized eggs across 20 fertilization replicates. After controlling for any changes in volume through incubation, results showed that successfully fertilized eggs were significantly larger than the eggs that failed to achieve fertilization. Under conditions without sperm limitation, fertility was unaffected by egg size. Our findings therefore support Levitan's theory, demonstrating empirically that some element of egg size variation could be selected by fertilization demands under sperm limitation. However, further research on sperm limitation in natural spawnings is required to assess the selective importance of these results.

A review of techniques of induced abortion.

PubMed

Hepburn, S

1981-04-01

Various techniques are available for inducing abortion. Evacuation of the uterus through the vagina is generally the preferred method in first trimester pregnancies. Dilation of the cervical canal by inserting rod dilators or laminaria tents allows the withdrawal of the fetus. Suction procedures (vacuum aspiration, uterine aspiration, or suction curettage) are possible since the decidua are separable from the basal layer of endometrium. This removal by force does not damage other maternal tissue. A cannula is introduced into the uterine cavity through the dilated cervix and its operator is then connected to a pump by way of a flexible tube which delivers negative pressure of about 600 mm of mercury. When the fetus is withdrawn, the uterus is felt to contract onto the cannula. The average time for this procedure is 5 minutes. Surgical curettage or dilatation and evacuation first dilates the cervical canal and then removes fetal parts and tissue from ovum forceps; a sharp curette does the rest. Anesthesia for these procedures may be general, local, or spinal. The techniques of menstrual regulation is used before pregnancy can be confirmed. However with the advent of the RIA test for the beta subunit of HCG this procedure is rarely indicated. Induction of premature labor is used in the later 1/2 of the second trimester and utilizes prostaglandins. Intraamniobor usually begins within 24 hours. Hysterotomy and hysterectomy are surgical procedures used in abortions.

Glycogen distribution in porcine fallopian tube epithelium during the estrus cycle.

PubMed

Gregoraszczuk, E Ł; Cała, M; Witkowska, E

2000-01-01

Histochemical features of two different parts of the porcine Fallopian tube have been studied, with special reference to cyclic changes in the distribution of glycogen particles. Porcine Fallopian tubes were obtained from a local slaughterhouse. Slides were studied under light microscopy utilising histological and histochemical techniques. The most striking feature during the periovulatory stage of the estrus cycle was the occurrence of glycogen granules in the apical cytoplasm of epithelial cells in both the ampulla and isthmus of the Fallopian tubes. In the isthmus, cells containing numerous granules of polysaccharides aggregated into areas of different sizes were noted after ovulation. During the midluteal phase their number was minimal or were even absent. In the ampula typical extrusion of secretory granules and nuclei protruding into the tubal lumen was visible after ovulation. In the luteal phase a lot of nuclei protruded into the tubal lumen and some free in the lumen were noted. It is possible that glycogen in the preovulatory stage functions as a source of energy for ciliary movement and as a nourishment for the ovum. In the isthmus large number of aggregated glycogen particles was observed also after ovulation. In this stage of the cycle, numerous granules of polysaccharide aggregated in isthmus epithelium could be the major energy source for embriogenesis when the embryo travels down the Fallopian tubes, during the early cleavage stage.

[Clinical profile of cytomegalovirus (CMV) enterocolitis in acquired immunodeficiency syndrome].

PubMed

De Lima, D B; Fernandes, O; Gomes, V R; Da Silva, E J; De Pinho, P R; De Paiva, D D

2000-01-01

To determine the clinical profile of CMV colitis in AIDS patients, comparing clinical, endoscopic parameters and survival time between 2 groups of AIDS patients having chronic diarrhea. Group A being CMV colitis and group B without CMV colitis. 48 patients with diarrhea that lasted more than 30 days, being 27 in Group A and 21 in Group B, were studied. Age, risk factors, interval time between the diagnosis of HIV infection and the beginning of diarrhea, hematochesia, the endoscopic findings and life table in both groups, were analysed. All of them were diagnosed by stool culture and stools for ovum and parasites, along colonoscopy with biopsies. The unpaired t test was used to assess statistical significance of differences observed in the means of continuous and the chi-square with Yates correction for non-parametric variables. The survival curves were assessed by the Kaplan-Meier and the Mantel-Haenszel's tests. A P value of less than 0,05 was considered to indicate statistical significance. The mucosal lesions associated with the CMV infection are typically ulcerative on a background of hemorrhagic erythema 14 (51,8%) p < 0,01. The life table analysis disclosed shorter survival time in the CMV colitis group 0,005> P>0,001. The others studied data did not achieve statistical significance. AIDS patients with CMV colitis have a poorer long-term survival. Among the colonoscopic findings, ulcerations with hemorrhagic background were the most common lesions.

Enhance beef cattle improvement by embryo biotechnologies.

PubMed

Wu, B; Zan, L

2012-10-01

Embryo biotechnology has become one of the prominent high businesses worldwide. This technology has evolved through three major changes, that is, traditional embryo transfer (in vivo embryo production by donor superovulation), in vitro embryo production by ovum pick up with in vitro fertilization and notably current cloning technique by somatic cell nuclear transfer and transgenic animal production. Embryo biotechnology has widely been used in dairy and beef cattle industry and commercial bovine embryo transfer has become a large international business. Currently, many developed biotechnologies during the period from early oocyte stage to pre-implantation embryos can be used to create new animal breeds and accelerate genetic progression. Based on recent advances in embryo biotechnologies and authors current studies, this review will focus on a description of the application of this technology to beef cattle improvement and discuss how to use this technology to accelerate beef cattle breeding and production. The main topics of this presentation include the following: (i) how to increase calf production numbers from gametes including sperm and oocyte; (ii) multiple ovulation and embryo transfer breeding schemes; (iii) in vitro fertilization and intracytoplasm sperm injection in bovine; (iv) pronuclear development and transgenic animals; (v) sex selection from sperm and embryos; (vi) cloning and androgenesis; (vii) blastocyst development and embryonic stem cells; (viii) preservation of beef cattle genetic resources; and (ix) conclusions. © 2011 Blackwell Verlag GmbH.

Analysis of ciliary beat frequency and ovum transport ability in the mouse oviduct.

PubMed

Shi, Dongbo; Komatsu, Kouji; Uemura, Tadashi; Fujimori, Toshihiko

2011-03-01

The oviduct is important in reproduction where fertilization occurs, and the fertilized eggs are conveyed to the uterus. Multi-ciliated cells of the oviductal epithelium and muscle contractions are believed to generate this unidirectional flow. Although there are many studies in human oviducts, there are few reports on mouse oviductal ciliary movements where we can dissect underlying genetic programs. To study ciliary movements in the mouse oviduct, we exposed the ovary-side of the oviduct (infundibulum) longitudinally and recorded the ciliary beatings in a hanging drop preparation. We calculated the ciliary beat frequency (CBF) by automated image analysis and found that the average CBF was 10.9 ± 3.3 and 8.5 ± 2.5 Hz (±standard deviation) during the diestrus and estrus stages, respectively. Mapping of the CBF to multiple locations in the epithelium showed that the cilia beat regularly at a local level, but have a range of frequencies within the entire plane. We also observed ova with cumulus cells were transported to the uterus side by the opened oviduct at the diestrus and estrus stages. These results suggest that the ciliated cells of the infundibulum can generate unidirectional flows and are able to deliver ova by their ciliary activities despite their discordance in beating periodicity. © 2011 The Authors. Journal compilation © 2011 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.

Occurrence of ovarian follicular dominance during stimulation for IVM impacts usable blastocyst yield.

PubMed

Romero, Sergio; Pella, Ricardo; Escudero, Francisco; Pérez, Ygor; García, Mario; Orihuela, Patricia

2018-03-01

To evaluate the influence of ovarian follicular dominance on the outcome of oocyte in-vitro maturation. This retrospective cohort study included 21 patients with polycystic ovaries or polycystic ovary syndrome (Rotterdam criteria, 2004) subjected to 24 invitro maturation (IVM) cycles between October 2015 and January 2017. Patients undergoing IVM received minimal gonadotropin stimulation starting on day 2 or 3 of the cycle; ovum pick-up typically occurred on days 6 to 8. No hCG-trigger shot was given. Following 30h of IVM, mature oocytes were inseminated by ICSI and the resulting embryos cultured up to the blastocyst stage. Ovarian follicular dominance was observed in nine of the 24 IVM cycles. Oocyte IVM yielded an overall maturation rate of 69.3±23.8%, and no difference was observed when the groups with or without a dominant follicle were assessed independently. The rates of fertilization and usable blastocysts per fertilized oocyte, mature oocyte (Metaphase II) or cumulus-oocyte-complex were nearly three times higher (28.7±22.5%) in the group without ovarian follicular dominance. No differences were found in the clinical pregnancy rates attained by the individuals with or without a dominant follicle after 21 vitrified-warmed blastocyst transfer cycles. Occurrence of ovarian follicular dominance during hormonal stimulation for in-vitro maturation negatively impacted embryological outcomes. Strategies devised to limit the appearance of ovarian follicular dominance must be further explored.

An analysis of 1150 cases of abortions from the Government R.S.R.M. Lying-in Hospital, Madras.

PubMed

Francis, O

1959-09-01

The Government R.S.R.M. Lying-in Hospital is located in one of the poorest sections of Madras, India, where the abortion rate is very high. The total number of complete abortions during the period, October 1957-November 1958, is 1150; the total number of deliveries including abortions is 10,367, an incidence rate of 11.09%. Of the 1150 cases, 789 (68.61%) were early abortions, up to 12 weeks; 361 (31.39%) were late, from the 13th to 28th week. An analysis of 1000 spontaneous abortions by Simons found that about 75% occurred before the 12th week. 758 abortions were performed on women aged 21-30; 204 occurred among those 31-40. 253 (22%) were primary abortions, i.e. the first pregnancy ended in an abortion and 897 (78%) were secondary abortions, i.e. there were 1 or more viable pregnancies before the abortion. Fetal death may be caused by abnormalities of the ovum, genital tract, or general maternal causes, or rare paternal causes. No cause could be found in 549 (47.74%) cases, but an associated abnormality was found in 601 (52.25%) cases. In 518 cases a single factor caused the abortion; in 83 cases more than a single etiological factor was found. There were 89 habitual aborters (7.74%). 19 of these were primary and 70 were secondary abortions.

In vitro development of OPU-derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen-thawed embryos after transfer.

PubMed

Isobe, Tomohiro; Ikebata, Yoshihisa; Do, Lanh Thi Kim; Tanihara, Fuminori; Taniguchi, Masayasu; Otoi, Takeshige

2015-07-01

The optimization of single-embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick-up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen-thawed embryos after a direct transfer. When two-cell-stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen-thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen-thawed embryos after transfer to recipients. © 2014 Japanese Society of Animal Science.

Amino acid carryover in the subzonal space of mouse fertilized ova affects subsequent transport kinetics.

PubMed

Rudraraju, Nirmala; Baltz, Jay M

2009-11-01

SummaryWe have investigated whether culture in glycine-containing medium affects subsequent glycine transport by the specific transport system, GLYT1, which is the sole glycine transporter in fertilized mouse ova. When fertilized ova were maintained for 6 h in culture with a physiological level of glycine (1 mM), subsequent transport of radiolabelled glycine was decreased by 40% compared with fertilized ova that had been maintained in glycine-free medium. Kinetic measurements showed that the apparent glycine affinity was decreased after culture with glycine (Km increased from 0.20 to 0.41 mM), but maximal transport rate was unchanged (similar Vmax of 20 and 23 fmol/fertilized ovum/min). These findings could have reflected activation of GLYT1 by prolonged substrate starvation, similar to some other amino acid transport systems. However, our findings were instead consistent with the alteration in glycine transport being due to trapping of glycine within the zona pellucida resulting in competitive transport inhibition even after ova were removed from glycine-containing media. First, even very brief exposures to glycine resulted in decreased subsequent glycine transport rates, with a maximal effect apparent within ~6 min. Second, extensive washing (at least six) reversed the effect. Third, the effect was absent when zona-free fertilized ova were used. Thus, it appears that components of the external environment of preimplantation embryos may continue to affect transport kinetics for a period even after embryos are removed from environments that contain them.

Assisted reproductive technology: Islamic Sunni perspective.

PubMed

Chamsi-Pasha, Hassan; Albar, Mohammed Ali

2015-06-01

Islam acknowledges that infertility is a significant hardship. Attempts to cure infertility are not only permissible, but also encouraged in Islam. Over the last three decades, a multitude of advances in assisted reproductive technologies (ARTs) have appeared. This review was carried out to inform readers, who are not familiar with Islamic doctrine, about the Sunni perspective on this topic. Systematic review of the literature. A series of searches was conducted of Medline databases published in English between January 1978 and December 2013 with the following assisted reproduction, infertility, gender selection, ethics, bioethics, and Islam. In Islamic Sunni law, all ARTs are allowed, provided that the source of the sperm, ovum, and uterus comes from a legally married couple during the span of their marriage. All forms of surrogacy are forbidden. A third-party donor is not allowed, whether he or she is providing sperm, eggs, embryos, or a uterus. Frozen preimplantation may be transferred to the wife in a successive cycle provided the marital bondage is not absolved by death or divorce. Gender selection for medical reasons is permitted. It is allowed for limited social reasons by some jurists, provided it does not involve discrimination against either sex. ART is acceptable and commendable in Islamic Sunni law provided it is practiced within the husband and wife dyad during the span of their marital contract. No third party should intrude upon the marital function of procreation. Surrogacy is not accepted by Sunni Islamic authorities.

Analysis of prostate-specific antigen transcripts in chimpanzees, cynomolgus monkeys, baboons, and African green monkeys.

PubMed

Mubiru, James N; Yang, Alice S; Olsen, Christian; Nayak, Sudhir; Livi, Carolina B; Dick, Edward J; Owston, Michael; Garcia-Forey, Magdalena; Shade, Robert E; Rogers, Jeffrey

2014-01-01

The function of prostate-specific antigen (PSA) is to liquefy the semen coagulum so that the released sperm can fuse with the ovum. Fifteen spliced variants of the PSA gene have been reported in humans, but little is known about alternative splicing in nonhuman primates. Positive selection has been reported in sex- and reproductive-related genes from sea urchins to Drosophila to humans; however, there are few studies of adaptive evolution of the PSA gene. Here, using polymerase chain reaction (PCR) product cloning and sequencing, we study PSA transcript variant heterogeneity in the prostates of chimpanzees (Pan troglodytes), cynomolgus monkeys (Macaca fascicularis), baboons (Papio hamadryas anubis), and African green monkeys (Chlorocebus aethiops). Six PSA variants were identified in the chimpanzee prostate, but only two variants were found in cynomolgus monkeys, baboons, and African green monkeys. In the chimpanzee the full-length transcript is expressed at the same magnitude as the transcripts that retain intron 3. We have found previously unidentified splice variants of the PSA gene, some of which might be linked to disease conditions. Selection on the PSA gene was studied in 11 primate species by computational methods using the sequences reported here for African green monkey, cynomolgus monkey, baboon, and chimpanzee and other sequences available in public databases. A codon-based analysis (dN/dS) of the PSA gene identified potential adaptive evolution at five residue sites (Arg45, Lys70, Gln144, Pro189, and Thr203).

Regulation of HGF and c-MET Interaction in Normal Ovary and Ovarian Cancer.

PubMed

Kwon, Youngjoo; Godwin, Andrew K

2017-04-01

Binding of hepatocyte growth factor (HGF) to the c-MET receptor has mitogenic, motogenic, and morphogenic effects on cells. The versatile biological effects of HGF and c-MET interactions make them important contributors to the development of malignant tumors. We and others have demonstrated a therapeutic value in targeting the interaction of c-MET and HGF in epithelial ovarian cancer (EOC). However, both HGF and c-MET are expressed in the normal ovary as well. Therefore, it is important to understand the differences in mechanisms that control HGF signaling activation and its functional role in the normal ovary and EOC. In the normal ovary, HGF signaling may be under hormonal regulation. During ovulation, HGF-converting proteases are secreted and the subsequent activation of HGF signaling enhances the proliferation of ovarian surface epithelium in order to replenish the area damaged due to expulsion of the ovum. In contrast, EOC cells that exhibit epithelial characteristics constitutively express both c-MET and HGF-converting proteases such as urokinase-type plasminogen activator. In EOC, mechanisms to control the activation of HGF signaling are absent since HGF is provided locally from the tissue microenvironment as well as remotely throughout the body. Potential incessant HGF signaling in EOC may lead to an increase in proliferation, invasion through the stroma, and migration to other tissues of cancer cells. Therefore, targeting the interaction of c-MET and HGF would be beneficial in treating EOC.

Regulation of HGF and c-MET Interaction in Normal Ovary and Ovarian Cancer

PubMed Central

Kwon, Youngjoo; Godwin, Andrew K.

2016-01-01

Binding of hepatocyte growth factor (HGF) to the c-MET receptor has mitogenic, motogenic, and morphogenic effects on cells. The versatile biological effects of HGF and c-MET interactions make them important contributors to the development of malignant tumors. We and others have demonstrated a therapeutic value in targeting the interaction of c-MET and HGF in epithelial ovarian cancer (EOC). However, both HGF and c-MET are expressed in the normal ovary as well. Therefore, it is important to understand the differences in mechanisms that control HGF signaling activation and its functional role in the normal ovary and EOC. In the normal ovary, HGF signaling may be under hormonal regulation. During ovulation, HGF-converting proteases are secreted and the subsequent activation of HGF signaling enhances the proliferation of ovarian surface epithelium in order to replenish the area damaged due to expulsion of the ovum. In contrast, EOC cells that exhibit epithelial characteristics constitutively express both c-MET and HGF-converting proteases such as urokinase-type plasminogen activator. In EOC, mechanisms to control the activation of HGF signaling are absent since HGF is provided locally from the tissue microenvironment as well as remotely throughout the body. Potential incessant HGF signaling in EOC may lead to an increase in proliferation, invasion through the stroma, and migration to other tissues of cancer cells. Therefore, targeting the interaction of c-MET and HGF would be beneficial in treating EOC. PMID:27170665

Impact of egg donation deliveries from domestic and overseas sources on maternal care: a questionnaire survey of Japanese perinatal physicians.

PubMed

Hibino, Yuri; Shimazono, Yosuke

2014-07-01

Recent demographic changes, such as marriage at later ages and delayed childbearing, have contributed to the increased demand for ovum donation. The purpose of the present study was to evaluate the frequency and impact of egg donation deliveries on maternal care using a questionnaire survey of Japanese perinatal care providers. A quantitative survey was conducted from October to November 2012 using self-administered anonymous questionnaires. We asked 2,693 obstetrics clinics/hospitals throughout Japan to complete the survey: 679 questionnaires were returned (response rate, 25.2 %). Of the respondents, 15.8 % answered that they had handled egg donation deliveries in the past. With regards to the country in which patients received egg donation services, the most frequent was the United States, including Hawaii. Asian countries, such as Thailand, Korea, and Singapore, were also reported; only two cases in Japan were reported. "Advanced age/menopause" was the most frequent reason for egg donation, and the mean age at egg donation delivery, because of advanced age/menopause, was 48.3 years. Our findings will increase public awareness of the legal issues related to assisted reproductive technology and cross-border reproductive care, as well as care of the mother and child in pregnancies resulting from reproductive technologies such as egg donation overseas. People should be aware of the issues involved in egg donation abroad and the resulting deliveries, and should implement specific care for women bearing children at later ages.

A natural stem cell therapy? How novel findings and biotechnology clarify the ethics of stem cell research

PubMed Central

Patel, P

2006-01-01

The natural replacement of damaged cells by stem cells occurs actively and often in adult tissues, especially rapidly dividing cells such as blood cells. An exciting case in Boston, however, posits a kind of natural stem cell therapy provided to a mother by her fetus—long after the fetus is born. Because there is a profound lack of medical intervention, this therapy seems natural enough and is unlikely to be morally suspect. Nevertheless, we feel morally uncertain when we consider giving this type of therapy to patients who would not naturally receive it. Much has been written about the ethics of stem cell research and therapy; this paper will focus on how recent advances in biotechnology and biological understandings of development narrow the debate. Here, the author briefly reviews current stem cell research practices, revisits the natural stem cell therapy case for moral evaluation, and ultimately demonstrates the importance of permissible stem cell research and therapy, even absent an agreement about the definition of when embryonic life begins. Although one promising technology, blighted ovum utilisation, uses fertilised but developmentally bankrupt eggs, it is argued that utilisation of unfertilised eggs to derive totipotent stem cells obviates the moral debate over when life begins. There are two existing technologies that fulfil this criterion: somatic cell nuclear transfer and parthenogenic stem cell derivation. Although these technologies are far from therapeutic, concerns over the morality of embryonic stem cell derivation should not hinder their advancement. PMID:16574879

Dermatological Manifestations in Patients Undergoing In Vitro Fertilisation: A Prospective Study.

PubMed

Sood, Aradhana; Sahu, Suvash; Karunakaran, Sandeep; Joshi, Rajneesh K; Raman, Deep Kumar

Changing sociodemographic patterns with an increase in the age of childbirth have affected fertility rates worldwide. With advancing reproductive medicine, assisted reproductive techniques (ARTs) are becoming common. While dermatological manifestations in normal pregnancies have been well documented, there is a paucity of data regarding cutaneous manifestations in patients undergoing ART. The objectives of our study were to estimate the incidence and types of dermatological manifestations in patients undergoing in vitro fertilisation (IVF) and to study their associations with age, type of infertility, and outcome of the procedure. A prospective cohort of 200 patients undergoing IVF in a tertiary care centre was observed for occurrence of any dermatological manifestations from initiation of the IVF protocol to the outcome of the procedure at 3 weeks after embryo transfer. Dermatological manifestations were seen in 27% of the study group, with urticaria being the most common cutaneous finding seen in 13.5%, followed by acneform eruptions (3%). Twenty-six (96.3%) of patients who manifested with urticaria were on progesterone. No statistically significant association was found between the occurrence of dermatological manifestations and the outcome of IVF, type of infertility, history of ART, and ovum donation in our study. Association between the age of the patient and the outcome of IVF cycle was statistically significant. Dermatological manifestations are seen in almost one-quarter of patients undergoing IVF, with progesterone-induced urticaria being the most common. Occurrence of cutaneous manifestations has no significant association with the outcome of IVF.

Obesity-Induced Dysfunctions in Female Reproduction: Lessons from Birds and Mammals123

PubMed Central

Walzem, Rosemary L.; Chen, Shuen-ei

2014-01-01

Follicle wall rupture and ovum release, i.e., ovulation, has been described as a controlled inflammatory event. The process involves tissue remodeling achieved through leukocyte-mediated proteolysis. In birds, ovulation is the first step in the energy-intensive process of egg formation, yet hens that consume energy in excess of productive requirements experience impaired egg-laying ability. Broiler chickens, selected for rapid lean muscle gain, and coincidentally hyperphagia, develop adult obesity when given free access to feed. Obese broiler hens experience elevated circulating concentrations of insulin and leptin, changes in lipid and lipoprotein metabolism similar to those of human metabolic syndrome, as well as increased systemic inflammation. Overall, the manifestations in poultry are similar to those of women with polycystic ovary syndrome. It was shown recently that, in hens, as in mammals, changes in lipid synthesis and metabolism cause granulosa cell apoptosis and altered immune function and hormone production, further compromising ovarian function. To date, there is insufficient information on the means used by the ovary to direct leukocyte function toward successful ovulation. More information is needed regarding the control of proteolytic actions by leukocytes with regards to the roles of specific enzymes in both ovulation and atresia. The broiler hen has provided unique insight into the interrelations of energy intake, obesity, leukocyte function, and reproduction. Additional work with this model can serve the dual purposes of improving avian reproduction and providing novel insights into polycystic ovary syndrome in women. PMID:24618762

Occurrence of ovarian follicular dominance during stimulation for IVM impacts usable blastocyst yield

PubMed Central

Romero, Sergio; Pella, Ricardo; Escudero, Francisco; Pérez, Ygor; García, Mario; Orihuela, Patricia

2018-01-01

Objective To evaluate the influence of ovarian follicular dominance on the outcome of oocyte in-vitro maturation. Methods This retrospective cohort study included 21 patients with polycystic ovaries or polycystic ovary syndrome (Rotterdam criteria, 2004) subjected to 24 in-vitro maturation (IVM) cycles between October 2015 and January 2017. Patients undergoing IVM received minimal gonadotropin stimulation starting on day 2 or 3 of the cycle; ovum pick-up typically occurred on days 6 to 8. No hCG-trigger shot was given. Following 30h of IVM, mature oocytes were inseminated by ICSI and the resulting embryos cultured up to the blastocyst stage. Results Ovarian follicular dominance was observed in nine of the 24 IVM cycles. Oocyte IVM yielded an overall maturation rate of 69.3±23.8%, and no difference was observed when the groups with or without a dominant follicle were assessed independently. The rates of fertilization and usable blastocysts per fertilized oocyte, mature oocyte (Metaphase II) or cumulus-oocyte-complex were nearly three times higher (28.7±22.5%) in the group without ovarian follicular dominance. No differences were found in the clinical pregnancy rates attained by the individuals with or without a dominant follicle after 21 vitrified-warmed blastocyst transfer cycles. Conclusion Occurrence of ovarian follicular dominance during hormonal stimulation for in-vitro maturation negatively impacted embryological outcomes. Strategies devised to limit the appearance of ovarian follicular dominance must be further explored. PMID:29338139

Diagnostic dilemma in ovarian pregnancy: a case series.

PubMed

Begum, Jasmina; Pallavee, P; Samal, Sunita

2015-04-01

Ovarian pregnancy is a rare form of ectopic pregnancy but it is the most common type of nontubal ectopic pregnancy. Many times it is operated with a misdiagnosis of ruptured tubal ectopic pregnancy or hemorrhagic corpus luteum. The high resolution transvaginal ultrasonography is a valuable tool for diagnosis of ectopic pregnancy but ovarian pregnancy still remains a diagnostic problem and a continuous challenge to the gynecologist. The correct diagnosis is made at the time of surgery and confirmation is by histopathological report. Here we report three cases of primary ovarian ectopic pregnancies, consistent with the Spiegelberg's criteria. Out of this, two cases have corroboration of ovarian ectopic pregnancy with use of intrauterine contraceptive device and one case by chance without any preexisting risk factors, probably due to interference in the release of ovum from the follicle. In all the three cases, emergency laparotomy was done for ruptured tubal ectopic pregnancy and the diagnosis of ruptured primary ovarian pregnancy was made at the time of surgery, this was subsequently confirmed by histopathology report. In the era where wider usage of intrauterine devices, ovulatory drugs and assisted reproductive techniques are rife, there is a possibility of an increase in the incidence of this rare entity, so ovarian ectopic pregnancy should be kept in mind as a possibility. Thereby early diagnosis by high resolution transvaginal ultrasound and laparoscopy can decrease the risk of complications like rupture, secondary implantation, hemorrhagic shock and maternal mortality.

Sperm retention site and its influence on pronucleus stage evaluation following intracytoplasmic sperm injection.

PubMed

Negishi, Momoko; Yanaihara, Atsushi; Iwasaki, Shinji; Suzuki, Norio; Hasegawa, Junichi; Yorimitsu, Takeshi; Okai, Takashi

2007-09-01

Aim:  It has been suggested that the position of the sperm after intracytoplasmic sperm injection (ICSI) has an effect on the development and quality of the embryo. In this study, we retrospectively examined whether pronucleus stage evaluation used through clinical studies in recent years has relevance with regard to sperm location. Methods:  From 2003 to 2005, 1285 oocytes from 459 patients (average age: 36 years) were retrospectively analyzed. The 459 patients underwent ICSI because of fertilization disorders and oligozoospermia. Follicle stimulation was via either Clomid or the long protocol. Human chorionic gonadotropin was administered to induce ovulation and oocyte retrieval was conducted 35 h later. After confirming the presence of a polar body, we immobilized the ovum at the 6 o'clock position, introduced the injection pipette at the 3 o'clock position and carried out ICSI. Results:  When a sperm was located at a position that was opposite to the polar body, both classifications of Scott and Tesarik regarding embryo quality were distinctly low. Furthermore, a good embryo classification ensued when the sperm was located adjacent to the polar body. Conclusion:  The zone in which the sperm was located did not always correlate with embryo quality; however, our study suggested that sperm location affects the synchronization of the nucleolus. When carrying out ICSI, it is important to take into consideration the insertion point of the sperm. (Reprod Med Biol 2007; 6 : 171-174).

Ovarian preservation with subcutaneous transposition in the setting of cytoreductive surgery

PubMed Central

Tan, Winson Jianhong; Jung, JingJin; Chia, Claramae Shulyn; Teo, Melissa Ching Ching; Toh, Han Chong; Soo, Khee Chee

2013-01-01

INTRODUCTION In recent years, cytoreductive surgery (CRS) with hyperthermic intraperitoneal chemotherapy (HIPEC) has gained increasing acceptance as a treatment modality for peritoneal carcinomatosis. In female patients, this procedure involves a total hysterectomy and bilateral saphingo-oophorectomy to remove the pelvic peritoneum. We present a case of an unfortunate female adolescent with peritoneal carcinomatosis who underwent cytoreductive surgery and HIPEC. In view of the compelling circumstance, an innovative surgical technique was used to attempt ovarian preservation. PRESENTATION OF CASE A 14 year old girl with carcinoma of the sigmoid colon and peritoneal metastases was offered cytoreductive surgery and hyperthermic intra-peritoneal chemotherapy. In view of her age, ovarian preservation with subcutaneous transposition was performed during cytoreductive surgery. She is currently well 6 months post surgery and has resumed normal menstruation. We review the literature regarding ovarian preservation with subcutaneous transposition and discuss its benefit in pre-menopausal women undergoing peritonectomy and cytoreductive surgery for peritoneal carcinomatosis. DISCUSSION Subcutaneous transposition of the ovary in pre-menopasual patients requiring cytoreductive surgery spares them the sequelae of surgical castration. The subcutaneous location of the transposed ovary conveys advantages such as the ease of ultrasound surveillance and removal in event of disease recurrence. It also retains the possibility of future conception as the transposed ovary can easily be accessed for ovum extraction with assisted reproductive techniques. CONCLUSION Ovarian preservation with subcutaneous transposition is a technique worth considering in the treatment of pre-menopausal women who require cytoreductive surgery for peritoneal carcinomatosis. PMID:23396395

A comparison of the Cook single lumen immature ovum IVM needle to the Steiner-Tan pseudo double lumen flushing needle for oocyte retrieval for IVM.

PubMed

Rose, B I; Laky, D

2013-06-01

This study compared the impact of using the Steiner-Tan pseudo double lumen needle for antral follicle oocyte retrieval to using a conventional non-flushing needle. The Steiner-Tan needle has a much smaller dead space than the needles commonly used for IVM oocyte retrievals. This was a retrospective cohort study. The patient population was determined by the time period in which a patient underwent IVM in a single physician's IVF practice. The following data was abstracted from clinical and embryology records: oocytes retrieved, oocytes matured, early maturing oocytes, oocytes fertilized, embryo quality measures, retrieval time, needle punctures, clot formation, and clinical pregnancy rate. The Steiner-Tan needle did not increase the number of oocytes retrieved. It also did not increase the time required for retrieval. However, flushing of antral follicles significantly decreased clot formation in fluid aspirates. Use of the Steiner-Tan needle also significantly decreased the number of vaginal needle punctures during each case. There was a trend toward improved embryo quality, but statistical power was inadequate to show a difference. The primary benefit of the Steiner-Tan needle was on the embryological aspects of IVM. Decreased blood and blood clots in the aspirates made an IVM retrieval more like conventional IVF for the embryologist. The patient also experienced less tissue trauma without increasing anesthesia or surgical time. There was no improvement in the number of oocytes retrieved, but based on the results, we hypothesized that oocytes were more commonly retrieved from slightly large follicles than when using a routine needle.

Ammonia Inactivation of Ascaris Ova in Ecological Compost by Using Urine and Ash

PubMed Central

Parzen, Rebecca E.; Mercado Guzmán, Álvaro

2012-01-01

Viable ova of Ascaris lumbricoides, an indicator organism for pathogens, are frequently found in feces-derived compost produced from ecological toilets, demonstrating that threshold levels of time, temperature, pH, and moisture content for pathogen inactivation are not routinely met. Previous studies have determined that NH3 has ovicidal properties for pathogens, including Ascaris ova. This research attempted to achieve Ascaris inactivation via NH3 under environmental conditions commonly found in ecological toilets and using materials universally available in an ecological sanitation setting, including compost (feces and sawdust), urine, and ash. Compost mixed with stored urine and ash produced the most rapid inactivation, with significant inactivation observed after 2 weeks and with a time to 99% ovum inactivation (T99) of 8 weeks. Compost mixed with fresh urine and ash achieved a T99 of 15 weeks, after a 4-week lag phase. Both matrices had relatively high total-ammonia concentrations and pH values of >9.24 (pKa of ammonia). In compost mixed with ash only, and in compost mixed with fresh urine only, inactivation was observed after an 11-week lag phase. These matrices contained NH3 concentrations of 164 to 173 and 102 to 277 mg/liter, respectively, when inactivation occurred, which was below the previously hypothesized threshold for inactivation (280 mg/liter), suggesting that a lower threshold NH3 concentration may be possible with a longer contact time. Other significant results include the hydrolysis of urea to ammonia between pH values of 10.4 and 11.6, above the literature threshold pH of 10. PMID:22582051

Ammonia inactivation of Ascaris ova in ecological compost by using urine and ash.

PubMed

McKinley, James W; Parzen, Rebecca E; Mercado Guzmán, Álvaro

2012-08-01

Viable ova of Ascaris lumbricoides, an indicator organism for pathogens, are frequently found in feces-derived compost produced from ecological toilets, demonstrating that threshold levels of time, temperature, pH, and moisture content for pathogen inactivation are not routinely met. Previous studies have determined that NH(3) has ovicidal properties for pathogens, including Ascaris ova. This research attempted to achieve Ascaris inactivation via NH(3) under environmental conditions commonly found in ecological toilets and using materials universally available in an ecological sanitation setting, including compost (feces and sawdust), urine, and ash. Compost mixed with stored urine and ash produced the most rapid inactivation, with significant inactivation observed after 2 weeks and with a time to 99% ovum inactivation (T(99)) of 8 weeks. Compost mixed with fresh urine and ash achieved a T(99) of 15 weeks, after a 4-week lag phase. Both matrices had relatively high total-ammonia concentrations and pH values of >9.24 (pK(a) of ammonia). In compost mixed with ash only, and in compost mixed with fresh urine only, inactivation was observed after an 11-week lag phase. These matrices contained NH(3) concentrations of 164 to 173 and 102 to 277 mg/liter, respectively, when inactivation occurred, which was below the previously hypothesized threshold for inactivation (280 mg/liter), suggesting that a lower threshold NH(3) concentration may be possible with a longer contact time. Other significant results include the hydrolysis of urea to ammonia between pH values of 10.4 and 11.6, above the literature threshold pH of 10.

Prediction value of anti-Mullerian hormone (AMH) serum levels and antral follicle count (AFC) in hormonal contraceptive (HC) users and non-HC users undergoing IVF-PGD treatment.

PubMed

Bas-Lando, Maayan; Rabinowitz, Ron; Farkash, Rivka; Algur, Nurit; Rubinstein, Esther; Schonberger, Oshrat; Eldar-Geva, Talia

2017-10-01

Use of hormone contraceptives (HC) is very popular in the reproductive age and, therefore, evaluation of ovarian reserve would be a useful tool to accurately evaluate the reproductive potential in HC users. We conducted a retrospective cohort study of 41 HC users compared to 57 non-HC users undergoing IVF-preimplantation genetic diagnosis (PGD) aiming to evaluate the effect of HC on the levels of anti-Mullerian hormone (AMH), small (2-5 mm), large (6-10 mm) and total antral follicle count (AFC) and the ability of these markers to predict IVF outcome. Significant differences in large AFC (p = 0.04) and ovarian volume (p < 0.0001) were seen, however, there were no significant differences in small and total AFC or in serum AMH and FSH levels. Oocyte number significantly correlated with AMH and total AFC in HC users (p < 0.001) while in non-HC users these correlations were weaker. In HC users, the significant predictors of achieving <6 and >18 oocytes were AFC (ROC-AUC; 0.958, p = 0.001 and 0.883, p = 0.001) and AMH (ROC-AUC-0.858, p = 0.01 and 0.878, p = 0.001), respectively. The predictive values were less significant in non-HC users. These findings are important in women treated for PGD, in ovum donors and for assessing the fertility prognosis in women using HC and wishing to postpone pregnancy.

Production of the first cloned camel by somatic cell nuclear transfer.

PubMed

Wani, Nisar A; Wernery, U; Hassan, F A H; Wernery, R; Skidmore, J A

2010-02-01

In this study, we demonstrate the use of somatic cell nuclear transfer to produce the first cloned camelid, a dromedary camel (Camelus dromedarius) belonging to the family Camelidae. Donor karyoplasts were obtained from adult skin fibroblasts, cumulus cells, or fetal fibroblasts, and in vivo-matured oocytes, obtained from preovulatory follicles of superstimulated female camels by transvaginal ultrasound guided ovum pick-up, were used as cytoplasts. Reconstructed embryos were cultured in vitro for 7 days up to the hatching/hatched blastocyst stage before they were transferred to synchronized recipients on Day 6 after ovulation. Pregnancies were achieved from the embryos reconstructed from all cell types, and a healthy calf, named Injaz, was born from the pregnancy by an embryo reconstructed with cumulus cells. Genotype analyses, using 25 dromedary camel microsatellite markers, confirmed that the cloned calf was derived from the donor cell line and the ovarian tissue. In conclusion, the present study reports, for the first time, establishment of pregnancies and birth of the first cloned camelid, a dromedary camel (C. dromedarius), by use of somatic cell nuclear transfer. This has opened doors for the amelioration and preservation of genetically valuable animals like high milk producers, racing champions, and males of high genetic merit in camelids. We also demonstrated, for the first time, that adult and fetal fibroblasts can be cultured, expanded, and frozen without losing their ability to support the development of nuclear transfer embryos, a technology that may potentially be used to modify fibroblast genome by homologous recombination so as to generate genetically altered cloned animals.

Ovulation, Fertilization and Early Embryonic Development in the Menstruating Fruit Bat, Carollia perspicillata

PubMed Central

Rasweiler IV, John J.; Badwaik, Nilima K.; Mechineni, Kiranmayi V.

2010-01-01

To characterize periovulatory events, reproductive tracts were collected at 12 hr intervals from captive-bred, short-tailed fruit bats, Carollia perspicillata, on days 1-3 post coitum and examined histologically. Most bats bred readily. Graafian follicles developed large antra and exhibited preovulatory expansion of the cumulus oophorus. Ovulation had occurred in some on the morning, and in most by the evening, of day 1. The single ovum was released as a secondary oocyte and fertilized in the oviductal ampulla. Ovulated secondary oocytes were loosely associated with their cumulus cells, which were lost around the initiation of fertilization. Supernumerary spermatozoa were occasionally noted attached to the zonae pellucidae of oviductal ova, but never within the perivitelline space. By day 2, most ova had reached the pronuclear stage and by day 3, early cleavage stages. Several lines of evidence indicate that C. perspicillata is a spontaneous ovulator with a functional luteal phase. Most newly-mated females had recently-formed, but regressing corpora lutea, and thickened (albeit menstrual) uteri despite having been housed with males only for brief periods (< 23 days). Menstruation is usually periovulatory in this species. Furthermore, the interval between successive estrus periods in most mated females that failed to establish ongoing pregnancies at the first was 21 – 27 days. Menstruation involved substantial endometrial desquamation, plus associated bleeding, and generally extended to the evening of day 3, the last time point studied. In nearly all females with a recent corpus luteum (n=24/25; 96%), the preovulatory or newly-ruptured follicle was in the opposite ovary. PMID:21337714

Maturational changes in the survivability and fertility of fowl sperm during their passage through the male reproductive tract.

PubMed

Ahammad, Muslah U; Nishino, C; Tatemoto, H; Okura, N; Kawamoto, Y; Okamoto, S; Nakada, T

2011-10-01

The objective of this study was to examine whether domestic fowl (Gallus domesticus) sperm undergo maturation in their capacity for survival and fertilization in the male reproductive tract. Sperm collected from the testis, epididymis and the proximal, middle and distal vas deferens were simultaneously stored in vitro in minimum essential medium (MEM) at 39°C for 0, 3 and 6h, and at 4°C for 24 and 48h. Sperm membrane integrity was measured using the dual fluorescent stain SYBR-14/propidium iodide (PI). Aliquots of sperm from the various sites were subjected to artificial insemination (AI) into the uteri of hens to assess the duration of sperm survival in the oviduct and to determine the fertility status of the sperm. Testicular sperm exhibited a very low capacity to survive under in vitro liquid storage conditions, irrespective of the storage temperature used, and in the oviduct, and they had a low ability to fertilize the ovum. On the contrary, sperm from the distal vas deferens had a higher survival rate during in vitro storage periods, a longer life span in the oviduct, and high fertility. Survival and fertilizing capacity of the sperm recovered from the testes increased gradually (P<0.05) from the testes to the distal vas deferens. In conclusion, we suggest that fowl sperm may undergo functional maturation through a process of gradual changes in their survival and fertilization capacities during their passage through the successive parts of the male reproductive tract. Copyright © 2011 Elsevier B.V. All rights reserved.

Commodification of human tissue: implications for feminist and development ethics.

PubMed

Dickenson, Donna

2002-05-01

One effect of late capitalism--the commodification of practically everything--is to knock down the Chinese walls between the natural and productive realms, to use a Marxist framework. Women's labour in egg extraction and 'surrogate' motherhood might then be seen as what it is, labour which produces something of value. But this does not necessarily mean that women will benefit from the commodification of practically everything, in either North or South. In the newly developing biotechnologies involving stem cells, the reverse is more likely, particular given the the shortage in the North of the egg donors who will be increasingly necessary to therapeutic cloning. Although most of the ethical debate has focused on the status of the embryo, this is to define ethics with no reference to global or gender justice. There has been little or no debate about possible exploitation of women, particularly of ovum donors from the South. Countries of the South without national ethics committees or guidelines may be particularly vulnerable: although there is increasing awareness of the susceptibility of poorer countries to abuses in research ethics, very little has been written about how they might be affected by the enormously profitable new technologies exploiting human tissue. Even in the UK, although the new Medical Research Council guidelines make a good deal of the 'gift relationship', what they are actually about is commodification. If donors believe they are demonstrating altruism, but biotechnology firms and researchers use the discourse of commodity and profit, we have not 'incomplete commodification' but complete commodification with a plausibly human face.

Experimental embryology of mammals at the Jastrzebiec Institute of Genetics and Animal Breeding.

PubMed

Karasiewicz, Jolanta; Andrzej-Modlinski, Jacek

2008-01-01

Our Department of Experimental Embryology originated from The Laboratory of Embryo Biotechnology, which was organized and directed by Dr. Maria Czlonkowska until her premature death in 1991. Proving successful international transfer of frozen equine embryos and generation of an embryonic sheep-goat chimaera surviving ten years were outstanding achievements of her term. In the 1990s, we produced advanced fetuses of mice after reconstructing enucleated oocytes with embryonic stem (ES) cells, as well as mice originating entirely from ES cells by substitution of the inner cell mass with ES cells. Attempts at obtaining ES cells in sheep resulted in the establishment of embryo-derived epithelioid cell lines from Polish Heatherhead and Polish Merino breeds, producing overt chimaeras upon blastocyst injection. Successful re-cloning was achieved from 8-cell rabbit embryos, and healthy animals were born from the third generation of cloned embryos. Recently mice were born after transfer of 8-cell embryonic nuclei into selectively enucleated zygotes, and mouse blastocysts were produced from selectively enucleated germinal vesicle oocytes surrounded by follicular cells, upon their reconstruction with 2-cell nuclei and subsequent activation. Embryonic-somatic chimaeras were born after transfer of foetal fibroblasts into 8-cell embryos (mouse) and into morulae and blastocysts (sheep). We also regularly perform the following applications: in vitro production of bovine embryos from slaughterhouse oocytes or those recovered by ovum pick up; cryopreservation of oocytes and embryos (freezing: mouse, rabbit, sheep, goat; vitrification: rabbit, cow); and banking of somatic cells from endangered wild mammalian species (mainly Cervidae).

Time-lapse cinematography-compatible polystyrene-based microwell culture system: a novel tool for tracking the development of individual bovine embryos.

PubMed

Sugimura, Satoshi; Akai, Tomonori; Somfai, Tamás; Hirayama, Muneyuki; Aikawa, Yoshio; Ohtake, Masaki; Hattori, Hideshi; Kobayashi, Shuji; Hashiyada, Yutaka; Konishi, Kazuyuki; Imai, Kei

2010-12-01

We have developed a polystyrene-based well-of-the-well (WOW) system using injection molding to track individual embryos throughout culture using time-lapse cinematography (TLC). WOW culture of bovine embryos following in vitro fertilization was compared with conventional droplet culture (control). No differences between control- and WOW-cultured embryos were observed during development to the blastocyst stage. Morphological quality and inner cell mass (ICM) and trophectoderm (TE) cell numbers were not different between control- and WOW-derived blastocysts; however, apoptosis in both the ICM and TE cells was reduced in WOW culture (P < 0.01). Oxygen consumption in WOW-derived blastocysts was closer to physiological level than that of control-derived blastocysts. Moreover, WOW culture improved embryo viability, as indicated by increased pregnancy rates at Days 30 and 60 after embryo transfer (P < 0.05). TLC monitoring was performed to evaluate the cleavage pattern and the duration of the first cell cycle of embryos from oocytes collected by ovum pickup; correlations with success of pregnancy were determined. Logistic regression analysis indicated that the cleavage pattern correlated with success of pregnancy (P < 0.05), but cell cycle length did not. Higher pregnancy rates (66.7%) were observed for animals in which transferred blastocysts had undergone normal cleavage, identified by the presence of two blastomeres of the same size without fragmentation, than among those with abnormal cleavage (33.3%). These results suggest that our microwell culture system is a powerful tool for producing and selecting healthy embryos and for identifying viability biomarkers.

The effects of superovulation with gonadotropins on autoantibody levels in patients undergoing assisted reproductive cycles.

PubMed

Ashrafi, Mahnaz; Amirchaghmaghi, Elham; Arabipoor, Arezoo; Vesali, Samira; Salman-Yazdi, Reza

2018-04-25

To evaluate the effect of controlled ovarian stimulation (COH) with gonadotropins on the serum levels of autoantibodies in the women who underwent in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles and to compare basal levels of these autoantibodies between groups according to history of COH. This prospective cohort study was performed from October 2014 to March 2016 in the Royan Institute. The volunteered infertile women with regard to the inclusion criteria, who underwent IVF/ICSI cycles, were recruited. The COH was performed according to standard long GnRH agonist protocol. The mean levels of the autoantibodies including anti-nuclear, anti-smooth muscle, anti-ovarian, anti-mitochondrial, anti β 2 -glycoprotein I, anti-parietal cell and anti-follicle-stimulating hormone antibodies were measured at three time points: on the 3-5 days of the menstrual cycle, 1 week after starting of COH and the ovum pick-up (OPU) day. Of all participants (n = 189), 73 women had history of COH (group B) and 116 women did not have such history (group A). The analysis indicated that the autoantibodies changes during COH were similar in both groups. COH has no significant impact on the level of autoantibodies during the stimulation cycle. Multiple logistic regression analysis showed that the serum levels of anti-smooth muscle antibody on OPU day was the positive predictive factors for live birth following ART cycles in the studied population. No significant effect of COH on the studied autoantibodies by the time of OPU was found but further studies are required to interpret these results.

Reproductive physiology of the female Magellanic penguin (Spheniscus magellanicus): Insights from the study of a zoological colony.

PubMed

O'Brien, J K; Schmitt, T L; Nollens, H H; Dubach, J M; Robeck, T R

2016-01-01

Eight captive female Magellanic penguins (Spheniscus magellanicus) were monitored over a 10week period, commencing at 5weeks prior to egg lay (EL), to increase our understanding of the species' reproductive biology. Females in cordoned nest sites underwent cloacal artificial insemination (AI) every 4-7days with different semen donors for each insemination. The EL interval was 97.9±3.6h (range: 84-108h) and paternity analyses revealed that conceptive inseminations occurred from 11.5 to 4.5days before oviposition. A biphasic pattern of estradiol, testosterone, progesterone and the biochemical analytes triglyceride, iron, calcium and phosphorus occurred in relation to EL, with values increasing (P<0.05) to maximal concentrations during the three weeks preceding oviposition, then decreasing (P<0.05) rapidly after oviposition completion. In comparison with post-lay (baseline) values, concentrations of estradiol and testosterone relative to the first oviposition were elevated at Week-5, and those of triglyceride, a yolk formation index, as well as iron, calcium and phosphorus, became elevated at Week-4 (P<0.05). Collective data indicate an estimated total egg formation interval of 29days, with oviducal transit of the ovulated ovum occurring over the majority of the ∼4day EL interval. These findings indicate that egg formation is prolonged with folliculogenesis initiated at 5weeks or more prior to oviposition. Consequently, the period of folliculogenesis and egg formation is estimated to overlap with the final ∼3weeks that wild females spend at sea prior to returning to land for breeding. Copyright © 2015 Elsevier Inc. All rights reserved.

Procedure for maximizing oocyte harvest for in vitro embryo production in small ruminants.

PubMed

Gibbons, A; Pereyra Bonnet, F; Cueto, M I; Catala, M; Salamone, D F; Gonzalez-Bulnes, A

2007-08-01

Possible effects of repeated hormonal treatments and laparoscopic ovum pick-up (LOPU) on the efficiency of oocyte recovery rate and quality were determined in sheep and goats. In six adult Merino sheep and five Criolla goats, ovarian status was synchronized by a prostaglandin F(2 alpha) analogue and the insertion of an intravaginal sponge 48 h later. Follicle development was stimulated by a single dose of FSH (60 mg NIH-FSH-P1) plus a single dose of equine chorionic gonadotrophin (eCG; 300 UI). The first FSH/eCG doses were administered 48 h after the sponge insertion, being repeated every 4 days to complete a total of four treatments in sheep and three in goats. Follicles in both ovaries were categorized according to their diameter and follicular fluid was aspirated under laparoscopic observation without a vacuum pump. In sheep, during a 12-day-period, a total of 347 follicles were aspirated with a recovery rate of 46.9%. In goats, during an 8-day-period, 219 follicles were aspirated with a recovery rate of 45.6%. In both species, there were no significant differences in the number of aspirated follicles, oocyte recovery rate and good quality oocyte recovery rate. However, in sheep the oocyte recovery rate was higher for large follicles, whereas in goats no such effect was detected. In summary, current results indicate that retrieval of oocytes can be maximized, without affecting oocyte quality, by repeating 'oneshot' FSH/eCG regimes and LOPUs at intervals as short as 4 days.

Gonadal Mosaicism Induced by Chemical Treatment of Sperm in Drosophila melanogaster

PubMed Central

Lindsley, Dan L.; Hardy, Robert W.; Ripoll, Pedro; Lindsley, Dart

2016-01-01

Accurate interpretation of forward genetic screens of chromosomes exposed in mature spermatozoa to a mutagenic chemical requires understanding—incomplete to date—of how exposed chromosomes and their replicas proceed through early development stages from the fertilized ovum to establishment of the germline of the treated male’s offspring. We describe a model for early embryonic development and establishment of the germline of Drosophila melanogaster and a model-validating experiment. Our model proposes that, barring repair, DNA strands modified by treatment with alkylating agents are stable and mutagenic. Each replication of an alkylated strand can result in misreplication and a mutant-bearing daughter nucleus. Daughter nuclei thenceforth replicate faithfully and their descendants comprise the embryonic syncytium. Of the 256 nuclei present after the eighth division, several migrate into the polar plasm at the posterior end of the embryo to found the germline. Based upon distribution of descendants of the alkylated strands, the misreplication rate, and the number of nuclei selected as germline progenitors, the frequency of gonadal mosaicism is predictable. Experimentally, we tracked chromosomes 2 and 3 from EMS-treated sperm through a number of generations, to characterize autosomal recessive lethal mutations and infer gonadal genetic content of the sons of treated males. Over 50% of 106 sons bore germlines that were singly, doubly, or triply mosaic for chromosome 2 or chromosome 3. These findings were consistent with our model, assuming a rate of misreplication between 0.65 and 0.80 at each replication of an alkylated strand. Crossing treated males to mismatch-repair-deficient females had no apparent effect on mutation rate. PMID:26163187

Embryonic genotype and inbreeding affect preimplantation development in cattle.

PubMed

Lazzari, G; Colleoni, S; Duchi, R; Galli, A; Houghton, F D; Galli, C

2011-05-01

Infertility in cattle herds is a growing problem with multifactorial causes. Embryonic genotype and level of inbreeding are among the many factors that can play a role on reproductive efficiency. To investigate this issue, we produced purebred and crossbred bovine embryos by in vitro techniques from Holstein oocytes and Holstein or Brown Swiss semen and analyzed several cellular and molecular features. In the first experiment, purebred and crossbred embryos, obtained from abattoir oocytes, were analyzed for cleavage, development to morula/blastocyst stages, amino acid metabolism and gene expression of developmentally important genes. The results indicated significant differences in the percentage of compacted morulae, in the expression of three genes at the blastocyst stage (MNSOD, GP130 and FGF4) and in the utilization of serine, asparagine, methionine and tryptophan in day 6 embryos. In the second experiment, bovine oocytes were collected by ovum pick up from ten Holstein donors and fertilized with the semen of the respective Holstein sires or with Brown Swiss semen. The derived embryos were grown in vitro up to day 7, and were then transferred to synchronized recipients and recovered on day 12. We found that purebred/inbred embryos had lower blastocyst rate on days 7-8, were smaller on day 12 and had lower expression of the trophoblast gene PLAC8. Overall, these results indicate reduced and delayed development of purebred embryos compared with crossbred embryos. In conclusion, this study provides evidence that embryo genotype and high inbreeding can affect amino acid metabolism, gene expression, preimplantation development and therefore fertility in cattle.

IVF culture medium affects human intrauterine growth as early as the second trimester of pregnancy.

PubMed

Nelissen, Ewka C M; Van Montfoort, Aafke P A; Smits, Luc J M; Menheere, Paul P C A; Evers, Johannes L H; Coonen, Edith; Derhaag, Josien G; Peeters, Louis L; Coumans, Audrey B; Dumoulin, John C M

2013-08-01

When does a difference in human intrauterine growth of singletons conceived after IVF and embryo culture in two different culture media appear? Differences in fetal development after culture of embryos in one of two IVF media were apparent as early as the second trimester of pregnancy. Abnormal fetal growth patterns are a major risk factor for the development of chronic diseases in adult life. Previously, we have shown that the medium used for culturing embryos during the first few days after fertilization significantly affects the birthweight of the resulting human singletons. The exact onset of this growth difference was unknown. In this retrospective cohort study, all 294 singleton live births after fresh embryo transfer in the period July 2003 to December 2006 were included. These embryos originated from IVF treatments that were part of a previously described clinical trial. Embryos were allocated to culture in either Vitrolife or Cook commercially available sequential culture media. We analysed ultrasound examinations at 8 (n = 290), 12 (n = 83) and 20 weeks' (n = 206) gestation and used first-trimester serum markers [pregnancy-associated plasma protein-A (PAPP-A) and free β-hCG]. Differences between study groups were tested by the Student's t-test, χ(2) test or Fisher's exact test, and linear multivariable regression analysis to adjust for possible confounders (for example, parity, gestational age at the time of ultrasound and fetal gender). A total of 294 singleton pregnancies (Vitrolife group nVL = 168, Cook group: nC = 126) from 294 couples were included. At 8 weeks' gestation, there was no difference between crown-rump length-based and ovum retrieval-based gestational age (ΔGA) (nVL = 163, nC = 122, adjusted mean difference, -0.04 days, P = 0.84). A total of 83 women underwent first-trimester screening at 12 weeks' gestation (nVL = 45, nC = 38). ΔGA, nuchal translucency (multiples of median, MoM) and PAPP-A (MoM) did not differ between the study groups. Free β-hCG (MoM) ± SEM differed significantly (1.55 ± 0.19 in Vitrolife versus 1.06 ± 0.10 in Cook; P = 0.031, Student's t-test). At 20 weeks' gestation, a more advanced GA, reflecting an increased fetal growth, was seen at ultrasound examination in the Vitrolife group (n = 115) when compared with the Cook group (n = 91). After adjustment for confounding factors, both the difference between GA based on three biparietal diameter dating formulas minus the actual (ovum retrieval based) GA (adjusted mean difference + 1.14 days (P = 0.04), +1.14 days (P = 0.04) and +1.36 days (P = 0.048)), as well as head circumference (HC) and trans-cerebellar diameter (TCD) were significantly higher in the Vitrolife group (HCvl 177.3 mm, HCc 175.9 mm, adjusted mean difference 1.8, P = 0.03; TCDvl 20.5 mm, TCDc 20.2 mm, adjusted mean difference 0.4, P = 0.008). A first trimester (12 weeks) fetal screening was not yet offered routinely during the study period, therefore only 28% of women in our study participated in this elective screening programme. Although all sonographers were experienced and specially trained to perform these ultrasound examinations and were unaware of the randomization procedure, we cannot totally rule out possible intra- and inter-observer variability. Despite being indispensable in daily practice, sonographic weight formulas have a limited accuracy. According to the fetal origins hypothesis, many adult diseases originate in utero owing to adaptations made by the fetus to the environment it encounters. This study indicates that the embryonic environment is already important for fetal development. Therefore, our study emphasizes the need to investigate fetal growth patterns after assisted reproduction technologies and long-term health outcomes of IVF children, especially in relation to the culture medium used during the first few days of preimplantation development. Not applicable.

Changes in affect and state anxiety across an in vitro fertilization/intracytoplasmic sperm injection cycle.

PubMed

Mahajan, Neha N; Turnbull, Deborah A; Davies, Michael J; Jindal, Umesh N; Briggs, Nancy E; Taplin, John E

2010-02-01

To identify pattern of change in average positive affect (PA), negative affect (NA), and state anxiety (St ANX) across three biological end points of an IVF/intracytoplasmic sperm injection (ICSI) procedure and to examine whether the pattern varied across sociodemographic and biomedical subgroups. Longitudinal follow-up study of PA, NA, and St ANX at three different time points: before start of study, before ovum pick-up (OPU), and before embryo transfer. Three infertility centers in northern India. Baseline data were obtained from a consecutive sample of 85 women. However, final analysis was done on data obtained from 74 women who reached the embryo transfer stage and completed the questionnaires at both OPU and embryo transfer. The PA, NA, and St ANX scores. Change in PA, NA, and St ANX scores at three stages of the treatment: baseline (T(0)), before OPU (T(1)), before embryo transfer (T(2)). The PA scores before OPU and embryo transfer were significantly lower than those at baseline. The mean NA and St ANX scores before OPU and embryo transfer were significantly higher than baseline scores. Furthermore, mean NA before embryo transfer was significantly higher than mean NA before OPU. The PA and St ANX scores showed statistically insignificance within cycle variations. Furthermore, the mean PA and St ANX for a subgroup of women who reported more than moderate level of burden were less variable. The OPU and embryo transfer stages are more stressful than the baseline stage for most women across various sociodemographic and biomedical subgroups. Women with more than a moderate level of financial burden were relatively more stable. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Effect of Quyu Jiedu granule on microenvironment of ova in patients with endometriosis.

PubMed

Lian, Fang; Li, Xin-Ling; Sun, Zhen-Gao; Zhang, Jian-Wei; Liu, Yan-He; Ma, Feng-Mei

2009-02-01

To observe the effect of Quyu Jiedu Granules (, QJG) on the micro- microenvironment of ova in patients with endometriosis (EM). environment Twenty EM patients who received in vitro fertilization and embryo transfer (IVF-ET) were randomized equally into a treated group and a control group. Further, 20 patients who received IVF-ET due to oviduct factors were enrolled into a non-endometriosis group. The dosage of gonadotrophic hormone used, the number of ova attained, fertilization rate and clinical pregnancy rate were all observed, and the levels of tumor necrosis factor alpha (TNF-right harpoon over left harpoon) and interleukin 6 (IL-6) in follicular fluid as well as their mRNA expressions in ovarian granular cells were detected by RT-PCR on the very day of ovum attainment. The ova attainment (13.80+/-6.87) and fertilization rate (0.69+/-0.31) in the treated group were all higher than the corresponding values in the control group (9.80+/-5.32 and 0.47+/-0.22); the follicular fluid contents of TNF-alpha and IL-6 in the treated group were 1.38+/-0.21 ng/mL and 130.56+/-12.81 pg/mL, respectively, which were lower than those in the control group (1.98+/-0.34 ng/mL and 146.83+/-17.65 pg/mL, respectively). Further, the treated group showed much lower mRNA expressions of TNF-alpha and IL-6 in ovarian granular cells. The elevation of TNF-alpha and IL-6 contents in follicular fluid and their mRNA expressions in ovarian granular cells are possibly related to the low quality of ova in EM; QJG might raise the ova quality by reducing TNF-alpha and IL-6 levels to improve the living micro-environment for the ova.

Longitudinal Study of Reproductive Performance of Female Cattle Produced by Somatic Cell Nuclear Transfer

PubMed Central

Polejaeva, Irina A.; Broek, Diane M.; Walker, Shawn C.; Zhou, Wenli; Walton, Mark; Benninghoff, Abby D.; Faber, David C.

2013-01-01

The objective of this study was to determine whether or not reproductive performance in cattle produced by somatic cell nuclear transfer (SCNT) is significantly different from that of their genetic donors. To address this question, we directed two longitudinal studies using different embryo production procedures: (1) superovulation followed by artificial insemination (AI) and embryo collection and (2) ultrasound-guided ovum pick-up followed by in vitro fertilization (OPU-IVF). Collectively, these two studies represent the largest data set available for any species on the reproductive performance of female clones and their genetic donors as measured by their embryo production outcomes in commercial embryo production program. The large-scale study described herein was conducted over a six-year period of time and provides a unique comparison of 96 clones to the 40 corresponding genetic donors. To our knowledge, this is the first longitudinal study on the reproductive performance of cattle clones using OPU-IVF. With nearly 2,000 reproductive procedures performed and more than 9,200 transferable embryos produced, our observations show that the reproductive performance of cattle produced by SCNT is not different compared to their genetic donors for the production of transferable embryos after either AI followed by embryo collection (P = 0.77) or OPU-IVF (P = 0.97). These data are in agreement with previous reports showing that the reproductive capabilities of cloned cattle are equal to that of conventionally produced cattle. In conclusion, results of this longitudinal study once again demonstrate that cloning technology, in combination with superovulation, AI and embryo collection or OPU-IVF, provides a valuable tool for faster dissemination of superior maternal genetics. PMID:24391930

Ostracoda (Crustacea) association and a new species (Dolerocypris anatolia nov. sp.) from the Pliocene-Pleistocene Afşin-Elbistan (Kahraman Maraş) Coal Basin of Turkey

NASA Astrophysics Data System (ADS)

Tunoǧlu, Cemal; Besbelli, Berk; Ertekin, İbrahim Kadri

2012-04-01

The Afşin-Elbistan Coal Basin, which is one of the largest and most important Pliocene-Pleistocene lignite basins of Turkey, is located in Eastern Anatolia. The basin was formed between two normal faults having NE-SW direction and these faults controlled both the sedimentation and the subsidence. The coal horizon of over 50 meters in thickness indicates the balance between the sedimentation and subsidence rates, and was preserved during peat deposition. Coals were generated in this extensive and shallow freshwater lake and evolved from the Pliocene to Pleistocene. Typical faunal and floral assemblages of this ancient Afşin-Elbistan freshwater lake are Ostracoda, Mollusca (Gastropoda and Pelecypoda), spore-pollen and Characeae (gyrogonites). Eleven Cypridoidea species were identified from the investigation area. Eight of them are already known (Candona neglecta Sars, Candona iliensis Mandelstam, Candona aff. candida (Müller), Pseudocandona compressa (Koch), Cyclocypris ovum (Jurine), Ilyocypris gibba (Ramdohr), Cypris pubera Müller, Heterocypris salina (Brady)), whereas three belong to open nomenclature — Candona sp. and Eucypris sp.; Dolerocypris anatolia nov. sp. is proposed as a new species. Dolerocypris Kaufmann is one of the largest genera among the freshwater Ostracoda. It has a very wide geographical distribution. Representatives of this genus are actively swimming species found in shallow zones of freshwater lakes and reported from small grassy water bodies with megascopic plants. Dolerocypris anatolia nov. sp. is recorded from core samples of the Pliocene-Pleistocene Afşin-Elbistan Coal Basin for the first time.

Functional anatomy of the female genital organs of the wild black agouti (Dasyprocta fuliginosa) female in the Peruvian Amazon.

PubMed

Mayor, P; Bodmer, R E; Lopez-Bejar, M

2011-02-01

This study examined anatomical and histological characteristics of genital organs of 38 black agouti females in the wild in different reproductive stages, collected by rural hunters in the North-eastern Peruvian Amazon. Females in the follicular phase of the estrous cycle had greater antral follicle sizes than other females, the largest antral follicle measuring 2.34mm. Antral follicles in pregnant females and females in luteal phase of the estrous cycle had an average maximum diameter smaller than 1mm. In black agouti females in follicular phase, some antral follicles are selected to continue to growth, reaching a pre-ovulatory diameter of 2mm. Mean ovulation rate was 2.5 follicles and litter size was 2.1 embryos or fetuses per pregnant female, resulting in a rate of ovum mortality of 20.8%. Many follicles from which ovulation did not occur of 1-mm maximum diameter luteinize forming accessory CL. The constituent active luteal tissues of the ovary are functional and accessory CL. Although all females had accessory CL, transformation of follicles into accessory CL occurred especially in pregnant females, resulting in a contribution from 9% to 23% of the total luteal volume as pregnancy advances. The persistence of functional CL throughout pregnancy might reflect the importance for the maintenance of gestation and may be essential for the continuous hormonal production. The duplex uterus of the agouti female is composed by two completely independent uterine horns with correspondent separate cervices opening into the vagina. In pregnant females, most remarkable observed uterine adaptations were induced by the progressive enlargement caused by the normal pregnancy evolution. The wild black agouti showed different vaginal epithelium features in accordance with the reproductive state of the female. Copyright © 2011 Elsevier B.V. All rights reserved.

Epicutaneous sensitization with nematode antigens of fish parasites results in the production of specific IgG and IgE.

PubMed

Fontenelle, G; Knoff, M; Verícimo, M A; São Clemente, S C

2018-07-01

Fish consumption plays an important role in the human diet. Hoplias malabaricus, trahira, is a freshwater fish widely appreciated in several Brazilian states and it is frequently infected by Contracaecum multipapillatum third-instar larvae (L3). The aim of the present study was to evaluate the allergenic potential of the C. multipapillatum L3 crude extract (CECM). BALB/c mice were immunized intraperitoneally (ip) with 10 or 50 μg CECM associated with 2 mg of aluminium hydroxide on days 0, 14 and 48. The determination of specific IgG and IgE antibody levels was done after immunization, and the late immunity was evaluated by the intradermal reaction in the ear pavilion. Epicutaneous sensitization was performed in the dorsal region, with antigenic exposure via a Finn-type chamber, containing 100 μg of chicken ovum albumin (OVA) or 100 μg CECM. After the exposures, the specific antibody levels were determined. In the ip immunization, there was a gradual increase in IgG antibody levels, independent of CECM concentration. In relation to IgE production, it was transitory, and immunization with 10 μg was more efficient than that of 50 μg. The same result was observed in the cellular hypersensitivity reaction. In the case of antigen exposure by the epicutaneous route, it was verified that only CECM was able to induce detectable levels of specific IgG and IgE antibodies. In the present study it was demonstrated that both intraperitoneal immunization and epicutaneous contact with C. multipapillatum larval antigens are potentially capable of inducing allergic sensitization in mice.

Development of an assay for a biomarker of pregnancy and early fetal loss

DOE Office of Scientific and Technical Information (OSTI.GOV)

Canfield, R.E.; O'Connor, J.F.; Birken, S.

1987-10-01

Human chorionic gonadotropin (hCG) is a glycoprotein hormone, secreted by the syncytiotrophoblast cells of the fertilized ovum, that enters the maternal circulation at the time of endometrial implantation. It is composed of two nonidentical subunits; ..cap alpha.. and ..beta.., with molecular weights of 14 kD and 23 kD, respectively. Human chorionic gonadotropin binds to the same receptor as hLH and displays the same biological response, namely, to stimulate the declining function of the corpus luteum to produce progestins and estrogen late in the menstrual cycle. The differences in the structures of hCG and hLH have been exploited to develop antibodiesmore » that can measure hCG specifically in the presence of hLH. Two-site antibody binding assays have been developed, based on a surface immunological concept of hCG epitopes, that involve four distinct regions to which antibodies against hCG can bind simultaneously. Antibody cooperative effects, in conjunction with kinetic advantages derived from the concentration factors by use of the sandwich assay technique (immunoradiometric assay, IRMA), have enabled development of extremely sensitive and specific measurement protocols for urinary hCG. The assay described herein permits the detection of pregnancy on an average 25.4 days after the first day of the preceding menses, as opposed to 29.5 days for conventional radioimmunoassay techniques. In addition, the greater sensitivity and specificity of this assay method has permitted the detection of episodes of fetal loss not detected by radioimmunoassay of urine specimens. A large scale epidemiological study is in progress using this assay technique as a way to identify pregnancies that are lost before becoming clinically apparent.« less

Pregnancy rates for embryos transferred from early postpartum beef cows into recipients with normal estrous cycles.

PubMed

Schrick, F N; Inskeep, E K; Butcher, R L

1993-09-01

Survival rate of embryos from first ovulations of postpartum cows with SHORT (6.9 +/- 0.3 days; n = 35) or NORMAL (17.1 +/- 0.3 days; n = 42) luteal phases and quality of the embryos on Day 6 were compared. At 19 to 23 days postpartum, cows were allotted to receive a norgestomet implant for 9 days (normal luteal phase) or to serve as untreated controls (short luteal phase). Calves were weaned 7 days after initiation of treatment to induce behavioral estrus in cows for mating. In 25 cows, growth of the ovulatory follicle was monitored by ultrasonography. On Day 6 after estrus, embryos were recovered nonsurgically, and live embryos were transferred into recipient cows exhibiting normal estrous cycles. The medium used to flush the embryos from the uterus of each donor cow was assayed for prostaglandin F2 alpha (PGF2 alpha). Days from calf removal to estrus and size of ovulatory follicles at ovulation (4.1 +/- 0.3 days and 16.7 +/- 0.7 mm, respectively) did not differ between NORMAL and SHORT cows. Interval from detection of the ovulatory follicle to ovulation was longer in NORMAL (10 +/- 0.7 days) than in SHORT cows (8 +/- 0.6 days; p < 0.05). Rates of recovery of an embryo or ovum (64%), rates of fertilization (65%), and quality or stage of development of Day 6 embryos did not differ between SHORT and NORMAL cows. Overall pregnancy rate from recovered oocytes was 13% for SHORT and 32% for NORMAL cows (p = 0.06); survival of fertilized oocytes was 23% for SHORT and 47% for NORMAL cows (p = 0.08).(ABSTRACT TRUNCATED AT 250 WORDS)

Longitudinal study of reproductive performance of female cattle produced by somatic cell nuclear transfer.

PubMed

Polejaeva, Irina A; Broek, Diane M; Walker, Shawn C; Zhou, Wenli; Walton, Mark; Benninghoff, Abby D; Faber, David C

2013-01-01

The objective of this study was to determine whether or not reproductive performance in cattle produced by somatic cell nuclear transfer (SCNT) is significantly different from that of their genetic donors. To address this question, we directed two longitudinal studies using different embryo production procedures: (1) superovulation followed by artificial insemination (AI) and embryo collection and (2) ultrasound-guided ovum pick-up followed by in vitro fertilization (OPU-IVF). Collectively, these two studies represent the largest data set available for any species on the reproductive performance of female clones and their genetic donors as measured by their embryo production outcomes in commercial embryo production program. The large-scale study described herein was conducted over a six-year period of time and provides a unique comparison of 96 clones to the 40 corresponding genetic donors. To our knowledge, this is the first longitudinal study on the reproductive performance of cattle clones using OPU-IVF. With nearly 2,000 reproductive procedures performed and more than 9,200 transferable embryos produced, our observations show that the reproductive performance of cattle produced by SCNT is not different compared to their genetic donors for the production of transferable embryos after either AI followed by embryo collection (P = 0.77) or OPU-IVF (P = 0.97). These data are in agreement with previous reports showing that the reproductive capabilities of cloned cattle are equal to that of conventionally produced cattle. In conclusion, results of this longitudinal study once again demonstrate that cloning technology, in combination with superovulation, AI and embryo collection or OPU-IVF, provides a valuable tool for faster dissemination of superior maternal genetics.

Production of monozygotic twin calves using the blastomere separation technique and Well of the Well culture system.

PubMed

Tagawa, M; Matoba, S; Narita, M; Saito, N; Nagai, T; Imai, K

2008-03-15

The present study was conducted to establish a simple and efficient method of producing monozygotic twin calves using the blastomere separation technique. To produce monozygotic twin embryos from zona-free two- and eight-cell embryos, blastomeres were separated mechanically by pipetting to form two demi-embryos; each single blastomere from the two-cell embryo and tetra-blastomeres from the eight-cell embryo were cultured in vitro using the Well of the Well culture system (WOW). This culture system supported the successful arrangement of blastomeres, resulting in their subsequent aggregation to form a demi-embryo developing to the blastocyst stage without a zona pellucida. There was no significant difference in the development to the blastocyst stage between blastomeres separated from eight-cell (72.0%) and two-cell (62.0%) embryos. The production rates of the monozygotic pair blastocysts and transferable paired blastocysts for demi-embryos obtained from eight-cell embryos (64.0 and 45.0%, respectively) were higher than those for demi-embryos obtained from two-cell embryos (49.0 and 31.0%, P<0.05). The separated demi-embryos obtained from eight-cell embryos produced by IVM/IVF of oocytes collected by ovum pick-up (OPU) from elite cows and cultured in wells tended to have a higher pregnancy rate (78.9% vs. 57.1%) and similar monozygotic twinning rate (40.0% vs. 33.3%) compared with monozygotic twin blastocysts obtained by the conventional bisection of in vivo derived blastocysts. In conclusion, producing twins by separation of blastomeres in OPU-IVF embryos, followed by the WOW culture system, yielded viable monozygotic demi-embryos, resulting in high rates of pregnancy and twinning rates after embryo transfer.

Two kinds of potentiality: a critique of McGinn on the ethics of abortion.

PubMed

Jacquette, D

2001-01-01

In Moral Literacy, or How to Do the Right Thing, Colin McGinn proposes a consequentialist solution to the abortion dilemma. McGinn interprets moral rights and moral interests as attributable only to actually sentient beings by virtue of their ability to experience pleasure or pain. McGinn argues against the moral rights of potentially conscious human fetuses, on the grounds that the unjoined ova and spermatoza of any fertile men and women are also potentially sentient, but we do not generally suppose that unjoined human genetic germ plasm has moral rights. I argue that McGinn's reply equivocates between two different senses of 'potential sentience.' I distinguish between strong and weak potentiality, or between naturally probably potentiality and merely logically possible potentiality. I agree that it is reasonable to deny that a weak or merely logically possible potentially sentient fetus that would result from any unjoined ovum and sperm has a moral right to life. But I claim that this fact does not diminish the plausibility of extending a moral right or potential moral right to life to a naturally probable potentially sentient fetus, which we have good reason to believe will actually become sentient in the natural course of things if nothing is done to prevent its normal development. I conclude that it is not merely the potentiality, but the strong potentiality of a healthy, normally developing fetus that is soon to acquire sentience, moral interests, and, on McGinn's own terms, a moral right to life, that continues to sustain the abortion controversy, even among those who also want respect a woman's right to reproductive self-determination.

Prostasomes from four different species are able to produce extracellular adenosine triphosphate (ATP).

PubMed

Ronquist, K Göran; Ek, Bo; Morrell, Jane; Stavreus-Evers, Anneli; Ström Holst, Bodil; Humblot, Patrice; Ronquist, Gunnar; Larsson, Anders

2013-10-01

Prostasomes are extracellular vesicles. Intracellularly they are enclosed by another larger vesicle, a so called "storage vesicle" equivalent to a multivesicular body of late endosomal origin. Prostasomes in their extracellular context are thought to play a crucial role in fertilization. Prostasomes were purified according to a well worked-out schedule from seminal plasmas obtained from human, canine, equine and bovine species. The various prostasomes were subjected to SDS-PAGE separation and protein banding patterns were compared. To gain knowledge of the prostasomal protein systems pertaining to prostasomes of four different species proteins were analyzed using a proteomic approach. An in vitro assay was employed to demonstrate ATP formation by prostasomes of different species. The SDS-PAGE banding pattern of prostasomes from the four species revealed a richly faceted picture with most protein bands within the molecular weight range of 10-150kDa. Some protein bands seemed to be concordant among species although differently expressed and the number of protein bands of dog prostasomes seemed to be distinctly fewer. Special emphasis was put on proteins involved in energy metabolic turnover. Prostasomes from all four species were able to form extracellular adenosine triphosphate (ATP). ATP formation was balanced by ATPase activity linked to the four types of prostasomes. These potencies of a possession of functional ATP-forming enzymes by different prostasome types should be regarded against the knowledge of ATP having a profound effect on cell responses and now explicitly on the success of the sperm cell to fertilize the ovum. This study unravels energy metabolic relationships of prostasomes from four different species. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

Does intrauterine injection of low-molecular-weight heparin improve the clinical pregnancy rate in intracytoplasmic sperm injection?

PubMed Central

El-Faissal, Yahia; Aboulghar, Mona; Mansour, Ragaa; Serour, Gamal I; Aboulghar, Mohamed

2016-01-01

Objective Heparin can modulate proteins, and influence processes involved in implantation and trophoblastic development. This study aimed to assess the improvement of clinical pregnancy and implantation rates after local intrauterine injection of low-molecular-weight heparin (LMWH) in patients undergoing intracytoplasmic sperm injection (ICSI). Methods A randomised case/control design was followed in women scheduled for ICSI. The study arm was injected with intrauterine LMWH during mock embryo transfer immediately following the ovum pickup procedure, while the control arm was given an intrauterine injection with a similar volume of tissue culture media. Side effects, the clinical pregnancy rate, and the implantation rate were recorded. Results The pregnancy rate was acceptable (33.9%) in the LMWH arm with no significant reported side effects, confirming the safety of the intervention. No statistically significant differences were found in the clinical pregnancy and implantation rates between both groups (p=0.182 and p=0.096, respectively). The odds ratio of being pregnant after intrauterine injection with LMWH compared to the control group was 0.572 (95% confidence interval [CI], 0.27−1.22), while the risk ratio was 0.717 (95% CI, 0.46−1.13; p=0.146). No statistical significance was found between the two groups in other factors affecting implantation, such as day of transfer (p=0.726), number of embryos transferred (p=0.362), or embryo quality. Conclusion Intrauterine injection of LMWH is a safe intervention, but the dose used in this study failed to improve the outcome of ICSI. Based on its safety, further research involving modification of the dosage and/or the timing of administration could result in improved ICSI success rates. PMID:28090465

Cell Death and Reproductive Regression in Female Schistosoma mansoni

PubMed Central

Galanti, Sarah E.; Huang, Stanley Ching-Cheng; Pearce, Edward J.

2012-01-01

The vitellarium is a highly proliferative organ, producing cells which are incorporated along with a fertilized ovum into the schistosome egg. Vitellarial growth fails to occur in virgin female schistosomes in single sex (female-only) infections, and involution of this tissue, which is accompanied by physical shrinkage of the entire worm, occurs when mature females sexually regress upon removal from their male partners. We have found that upon removal from their hosts into tissue culture, female parasites regress whether they are mated or not, but that cessation of egg production and a decline in expression of the vitelline gene p14 is delayed by mating. We used BrdU labeling to investigate whether there was a loss of proliferation in the vittelarium that might account for regression and found that the proliferation rate declined equally in paired and singled females once placed into culture. However, TUNEL staining and Caspase 3 activity measurements indicate that the loss of vitrellarial cellularity associated with regression is associated with profound apoptotic vitelline cell death, which is not apparent in the vitellaria of paired females immediately ex vivo, and which develops in vitro regardless of whether males are present or not. Furthermore, primordial vitellaria in virgin females have a high frequency of apoptotic cells but are characterized by a proliferation rate that is indistinguishable from that in fully developed vitellaria in mature paired females. Taken together, our data suggest that the vitelline proliferation rate is independent of pairing status. In contrast, the survival of vitelline cells, and therefore the development of the vitellarium, is highly male-dependent. Both processes are negatively affected by removal from the host regardless of whether male worms are present or not, and are unsustainable using standard tissue culture approaches. PMID:22363825

Embryo transfer after 2 or 5 days of IVF culture: a retrospective comparison.

PubMed

Lundqvist, Monalill; Rova, Karin; Simberg, Niklas; Lundkvist, Orjan

2002-02-01

To determine whether prolongation of embryo culture in vitro from day 2 to day 5 after ovum pick-up (OPU) and fertilization can improve the results of in vitro fertilization (IVF), and the morphology of the spare embryos on day 2 can predict the developmental capacity during prolonged culture. We also wanted to consider this as a strategy to avoid twin pregnancies if it could be possible to transfer only one blastocyst at a time in the future. A retrospective analysis with embryo transfer timed according to the weekday of OPU. Embryo transfer was performed on day 2 in 103 cases and on day 5 in 120 cases. Only one cycle per couple was included. The pregnancy rates per embryo transfer on day 2 (27/103, 26%) and day 5 (36/120, 30%) were similar. There were significantly more miscarriages in the day 5 (50%) than in the day 2 group (22%, p = 0.02), but there was no significant difference in the baby take home rate (20% in day 2 group, 15% in day 5 group). The morphological appearance of the embryos on day 2 was poorly correlated to the developmental potential during prolonged culture in vitro. On day 5, transfer of one or two blastocysts resulted in a pregnancy rate that tended to be higher than that after transfer of morulae only. Prolongation of embryo culture from day 2 to day 5 did not improve the clinical outcome of the IVF treatment when measured as baby take home rate. Therefore, for the time being, this strategy does not increase our chances to move towards single embryo transfer.

Dynamic changes in leptin distribution in the progression from ovum to blastocyst of the pre-implantation mouse embryo

PubMed Central

Schulz, Laura C.; Roberts, R. Michael

2011-01-01

The hormone leptin, which is primarily produced by adipose tissue, is a critical permissive factor for multiple reproductive events in the mouse, including implantation. In the CD1 strain, maternally-derived leptin from the oocyte becomes differentially distributed among blastomeres of pre-implantation embryos to create a polarized pattern, a feature consistent with a model of development in which blastomeres are biased towards a particular fate as early as the 2-cell stage. Here, we have confirmed that embryonic leptin is of maternal origin and re-examined leptin distribution in two distinct strains in which embryos were derived after either normal ovulation or superovulation. A polarized pattern of leptin distribution was found in the majority of both CD1 and CF1 embryos (79.1 % and 76.9 %, respectively) collected following superovulation, but was reduced, particularly in CF1 embryos (29.8 %; p < 0.0001), after natural ovulation. The difference in leptin asymmetries in the CF1 strain arose between ovulation and the first cleavage division, and was not affected by removal of the zona pellucida. Presence or absence of leptin polarization was not linked to differences in ability of embryos to develop normally to blastocyst. In the early blastocyst, leptin was confined subcortically to trophectoderm but upon blastocoel expansion it was lost from cells. Throughout development leptin co-localized with LRP2, a multi-ligand transport protein, and its patterning resembled that noted for the maternal-effect proteins OOEP, NLRP5, and PADI6, suggesting that it is a component of the subcortical maternal complex with as yet unknown significance in pre-implantation development. PMID:21444625

Regulation of prohibitin expression during follicular development and atresia in the mammalian ovary.

PubMed

Thompson, Winston E; Asselin, Eric; Branch, Alicia; Stiles, Jonathan K; Sutovsky, Peter; Lai, Liangxue; Im, Gi-Sun; Prather, Randall S; Isom, S Clay; Rucker, Edmund; Tsang, Benjamin K

2004-07-01

Prohibitin is a ubiquitous and highly conserved protein implicated as an important regulator in cell survival. Prohibitin content is inversely associated with cell proliferation, but it increases during granulosa cell differentiation as well as in earlier events of apoptosis in a temperature-sensitive granulosa cell line. In the present study, we have characterized the spatial expression patterns for prohibitin using established in vivo models for the induction of follicular development and atresia in the mammalian ovary. Comparative Western blot analyses of granulosa cell lysates from control ovaries and from ovaries primed with eCG or treated with eCG plus anti-eCG (gonadotropin withdrawal) were conducted. Prohibitin was immunolocalized in rat ovarian sections probed with antibodies against either proliferating cell nuclear antigen (PCNA) or cholesterol side-chain cleavage cytochrome P450 (P450(scc)) or in terminal deoxynucleotidyl transferase-mediated dUTP nick end labeled sections. Additionally, porcine oocytes, zygotes, and blastocyts were also immunolocalized with prohibitin antibody. Immunolocalization revealed the presence of prohibitin in granulosa cells, theca-interstitial cells, and the oocyte. The results indicate that prohibitin protein expression in the gonadotropin-treated cells was upregulated. Immunoreactivity of prohibitin was inversely related to PCNA expression during follicular maturation and colocalized with P450(scc). Prohibitin appeared to be translocated from the cytoplasm to the nucleus in atretic follicles, germinal vesicle-stage oocytes, zygotes, and blastocysts. These results suggest that prohibitin has several functional regulatory roles in granulosa and theca-interstitial cells and in the ovum during follicular maturation and atresia. It is likely that prohibitin may play an important role in determining the fate of these cells and eventual follicular destiny.

Co-Administration of Metformin and N-Acetyl Cysteine Fails to Improve Clinical Manifestations in PCOS Individual Undergoing ICSI

PubMed Central

Cheraghi, Ebrahim; Soleimani Mehranjani, Malek; Shariatzadeh, Mohammad Ali; Nasr Esfahani, Mohammad Hossein; Ebrahimi, Zahra

2014-01-01

Background Studies have demonstrated the efficacy of metformin (MTF ) in reducing insulin resistance and N-acetyl cysteine (NAC) in inhibiting oxidative stress which are involved in the pathogenesis of polycystic ovarian syndrome (PCOS). We aimed to compare the effects of MTF and NAC combination on serum metabolite and hormonal levels during the course of ovulation induction in PCOS individual candidates of intracytoplasmic sperm injection (ICSI). Materials and Methods In this prospective randomized clinical trial, placebo con- trolled pilot study, 80 patients of polycystic ovarian syndrome at the age of 25-35 years were divided into 4 groups (n=20): i. NAC=treated with N-acetyl cysteine (600 mg three times daily), ii. MTF=treated with metformin (500 mg three times daily), iii. MTF+NAC=treated with N-acetyl cysteine plus metformin (the offered doses) and iv. placebo (PLA). A total number of 20 patients (6 from MTF group, 4 from NAC group, 6 from MTF+NAC group and 4 from PLA group) were dropped of the study. The drugs were administrated from day 3 of menses of previous cycle until ovum pick-up. Results Serum levels of luteinizing hormone (LH), total testosterone, cholester- ol and triglyceride, insulin and leptin significantly reduced in the MTF and NAC groups compared to the placebo (p<0.01). But levels of LH, total testosterone, cholesterol and triglyceride had no significant reduction in the MTF+NAC groups compared to the placebo. The serum levels of malonyldialdehyde (MDA), insulin and leptin reduced significantly after treatment in the MTF+NAC group compared to the placebo (p<0.05). Conclusion Considering the adverse effect of combination therapy, we proposed the conadministration might have no beneficial effect for PCOS patient during course of ovulation induction of ICSI (Registration Number: IRCT201204159476N1). PMID:25083175

Ultrasound-guided instrumental removal of the retained placenta after vaginal delivery.

PubMed

Rosenstein, Melissa G; Vargas, Juan E; Drey, Eleanor A

2014-08-01

The standard treatment for retained placenta is manual extraction, in which a hand is introduced inside the uterus to cleave a plane between the placenta and the uterine wall. For women without an epidural, the procedure is extremely uncomfortable and may require additional measures such as intravenous narcotics or regional anesthesia. Although ultrasound-guided instrumental removal of the placenta is standard practice as part of second-trimester abortion by dilation and evacuation and may be done at many institutions, especially after failed manual extraction, it has not yet been described in the literature as a technique following vaginal birth. Our experience with this technique is that it causes less discomfort to the patient than a traditional manual extraction, because the instrument entering the uterus is much narrower than a hand. With the patient in dorsal lithotomy, we locate the cervix and stabilize it either with fingers or a ring forceps on the anterior lip. We introduce Bierer ovum forceps into the uterus under direct ultrasound guidance. The Bierer forceps are preferred because of their long length, large head, and serrated teeth that allow for a firm, secure grip on the placenta. We grasp the placental tissue with the forceps and apply slow, gentle traction in short strokes, regrasping increasingly more distal areas of placenta as necessary to tease out the placenta. After 1-2 minutes, the placenta separates and can be pulled out of the uterus, usually intact. Our experience suggests that this technique is a well-tolerated option for women without an epidural who have a retained placenta. Further study is needed to quantify the amount of discomfort and anesthesia that can be avoided with this technique, as well as whether there is any change in the frequency of infectious complications or the necessity of postremoval curettage. Copyright © 2014 Mosby, Inc. All rights reserved.

Distribution and threshold expression of the tRNA[sup Lys] mutation in skeletal muscle of patients with myoclonic epilepsy and ragged-red fibers (MERRF)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boulet, L.; Karpati, G.; Shoubridge, E.A.

1992-12-01

The authors investigated the distribution and expression of mutant mtDNAs carrying the A-to-G mutation at position 8344 in the tRNA[sup Lys] gene in the skeletal muscle of four patients with myoclonus epilepsy and ragged-red fibers (MERRF). The proportion of mutant genomes was greater than 80% of total mtDNAs in muscle samples of all patients and was associated with a decrease in the activity of cytochrome c oxidase (COX). The vast majority of myoblasts, cloned from the satellite-cell population in the same muscles, were homoplasmic for the mutation. The overall proportion of mutant mtDNAs in this population was similar to thatmore » in differentiated muscle, suggesting that the ratio of mutant to wild-type mtDNAs in skeletal muscle is determined either in the ovum or during early development and changes little with age. Translation of all mtDNA-encoded genes was severely depressed in homoplasmic mutant myoblast clones but not in heteroplasmic or wild-type clones. The threshold for biochemical expression of the mutation was determined in heteroplasmic myotubes formed by fusion of different proportions of mutant and wild-type myoblasts. The magnitude of the decrease in translation in myotubes containing mutant mtDNAs was protein specific. Complex I and IV subunits were more affected than complex V subunits, and there was a rough correlation with both protein size and number of lysine residues. Approximately 15% wild-type mtDNAs restored translation and COX activity to near normal levels. These results show that the A-to-G substitution in tRNA[sup Lys] is a functionally recessive mutation that can be rescued by intraorganellar complementation with a small proportion of wild-type mtDNAs and explain the steep threshold for expression of the MERRF clinical phenotype. 40 refs., 7 figs., 2 tabs.« less

Charging the vas deferens -- a novel male contraceptive method.

PubMed

Koshy, L M

1994-08-30

A new male contraception method is undergoing clinical trials in India. The goal of this research is to achieve a method which restricts its effects to the region of the vas deferens. First, the belief that there was a strong peristaltic movement in the vas during ejaculation had to be examined. It was found that only the ampula part of the vas is given a tight squeeze to release the stored sperm into the semen. Therefore, a device to manipulate sperm could safely be inserted into the vas. Using the fact that the acrosomal membrane of the sperm is disturbed when sperm are subjected to electrical currents, Dr. S. K. Guha, of the Indian Institute of Technology in New Delhi, decided to try to generate a current inside the vas to disturb the acrosomal membranes. Since styrene maleic anhydride (SMA) hydrolyses when mixed with body fluids, it creates a slight charge. After injecting SMA into the vas, the electrical balance of the sperm which passes it is disturbed, causing the acrosomal membrane to shrink and rupture, letting water enter the sperm. When the sperm head bursts, the enzymes acrocin and hyaluronidase are released. Without these enzymes, the sperm can not penetrate the ovum and fertility control is achieved. This procedure can be reversed by injecting a solvent to dissolve the SMA in the scrotal sac. Phase I clinical studies began in 1989, and the next Phases should begin soon. The advantages of this method include the fact that no surgical procedure is required, sperm production is not actually blocked so there is no rise in anti-sperm antibody titres, and the contraceptive action can last as long as 10 years. Dr. Guha cautions, however, that education of the people will ultimately play a more important role in population control in India than the availability of a variety of contraceptive methods.

Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.

PubMed

Galli, C; Colleoni, S; Duchi, R; Lagutina, I; Lazzari, G

2007-03-01

Development of assisted reproductive technologies in horses has been relatively slow compared to other domestic species, namely ruminants and pigs. The scarce availability of abattoir ovaries and the lack of interest from horse breeders and breed associations have been the main reasons for this delay. Progressively though, the technology of oocyte maturation in vitro has been established followed by the application of ICSI to achieve fertilization in vitro. Embryo culture was initially performed in vivo, in the mare oviduct or in the surrogate sheep oviduct, to achieve the highest embryo development, in the range of 18-36% of the fertilised oocytes. Subsequently, the parallel improvement of in vitro oocyte maturation conditions and embryo culture media has permitted high rates of embryo development from in vitro matured and in vitro cultured ICSI embryos, ranging from 5 to 10% in the early studies to up to 38% in the latest ones. From 2003, with the birth of the first cloned equids, the technology of somatic cell nuclear transfer has also become established due to improvement of the basic steps of embryo production in vitro, including cryopreservation. Pregnancy and foaling rates are still estimated based on a small number of in vitro produced equine embryos transferred to recipients. The largest set of data on non-surgical embryo transfer of in vitro produced embryos, from ICSI of both abattoir and in vitro-matured Ovum Pick Up (OPU) oocytes, and from somatic cell nuclear transfer, has been obtained in our laboratory. The data demonstrate that equine embryos produced by OPU and then cryopreserved can achieve up to 69% pregnancy rate with a foaling rate of 83%. These percentages are reduced to 11 and 23%, respectively, for cloned embryos. In conclusion, extensive evidence exists that in vitro matured equine oocytes can efficiently develop into viable embryos and offspring.

Enhanced expressions of neurodegeneration-associated factors, UPS impairment, and excess Aβ accumulation in the hippocampus of mice with persistent cerebral toxocariasis.

PubMed

Chou, Chia-Mei; Lee, Yueh-Lun; Liao, Chien-Wei; Huang, Ying-Chieh; Fan, Chia-Kwung

2017-12-22

Toxocariasis is a worldwide zoonotic parasitic disease mainly caused by Toxocara canis. Humans can be infected by accidental ingestion of T. canis embryonated ovum-contaminated food, water, or encapsulated larvae in paratenic hosts' viscera or meat. Since humans and mice are paratenic hosts of T. canis, the wandering larvae might cause mechanical tissue damage and excretory-secretory antigens may trigger inflammatory injuries to local organs. Long-term residence of T. canis larvae in a paratenic host's brain may cause cerebral toxocariasis (CT) that contributes to cerebral damage, neuroinflammation and neuropsychiatric disorders in mice and clinical patients. Since the hippocampus has been long recognized as being responsible for learning and memory functions, parasitic invasion of this site may cause neuroinflammatory and neurodegenerative disorders. The present study intended to assess pathological changes, expressions of neurodegeneration-associated factors (NDAFs), including transforming growth factor (TGF)-β1, S100B, glial fibrillary acidic protein (GFAP), transglutaminase type 2 (TG2), claudin-5, substance P (SP) and interleukin (IL)-1β, and the ubiquitin-proteasome system (UPS) function in the hippocampus and associated cognitive behavior in ICR mice orally inoculated with a high, medium or low-dose of T. canis embryonated ova during a 20-week investigation. Results indicated although there were insignificant differences in learning and memory function between the experimental mice and uninfected control mice, possibly because the site where T. canis larvae invaded was the surrounding area but not the hippocampus per se. Nevertheless, enhanced expressions of NDAF, persistent UPS impairment and excess amyloid β (Aβ) accumulation concomitantly emerged in the experimental mice hippocampus at 8, 16 and 20 weeks post-infection. We thus postulate that progressive CT may still progress to neurodegeneration due to enhanced NDAF expressions, persistent UPS impairment and excess Aβ accumulation in the hippocampus.

Can the controversy about the putative role of the human female orgasm in sperm transport be settled with our current physiological knowledge of coitus?

PubMed

Levin, Roy J

2011-06-01

Spermatozoal uptake, facilitated by uterine contractions induced by oxytocin at orgasm during coitus, has been a long term concept. Studies attempting its support, however, have been poorly examined especially in the context of the changes in the female genital tract activated by sexual arousal. To examine experimental support for the concept. Using a variety of search engines, mainly peer reviewed articles and un-reviewed books were examined relating to sperm transport and function in the human female genital tract in the absence and presence of arousal to orgasm. Identifying evidence-based data to support authority-based opinion. All the experimental observations of sperm or model substitute's transport have been undertaken in women who were not sexually aroused. They fail to take into account that arousal creates vaginal tenting lifting the cervico-uterine complex into the false pelvis away from the ejaculated semen. This delays sperm uptake and transport making conclusions from these observations invalid in relation to transport during coitus. Studies injecting oxytocin have not used women in their sexually aroused state and used supraphysiological doses unlikely to be comparable with coitus and orgasm. The proposal that the transport of extra sperm by oxytocin-induced uterine contractions at orgasm is needed to facilitate fertility ignores possible harm from increased sperm numbers creating polyspermy and sperm enzyme release causing ovum degeneration, leading to decreased fertility. The role of sperm motility in their uptake from the vagina into the cervix as opposed to en bloc transfer through uterine archimyometrial-mediated transport in the absence of orgasm is at present unresolvable because of conflicting studies. The bulk of the reported evidence favors the conclusion that the female orgasm, with its concomitant central release of oxytocin, has little or no effective role in the transport of spermatozoa in natural human coitus. © 2010 International Society for Sexual Medicine.

Choosing GnRH Antagonist Protocol Shows Improved Oocyte and Embryo Quality, Coherent with the Perifollicular Vascularity (PFV) in Assisted Reproductive Techniques

PubMed Central

Ramachandran, Amar; Kumar, Pratap

2015-01-01

Introduction The parent oocyte from which the embryo is derived, determines its quality and the perifollicular vascularity (PFV) determines the micro-environment of the developing ovum. The PFV correlates well with the follicular oxygenation, oocyte maturation and embryo viability. PFV is imaged with Power Doppler Ultrasound. Aim To study and compare the association of the PFV of follicles with the quality of the oocytes and embryos in agonist and antagonist protocol, employed in Assisted Reproductive techniques (ART). Study Design A prospective observational study was conducted on 75 patients, who were recruited for ART cycles, out of which 25 were given the Agonist protocol and the remaining 50 received the Antagonist protocol. Materials and Methods The patients underwent the stimulation protocol. The PFV of preovulatory follicles were studied with Transvaginal Power Doppler and graded. Each oocyte retrieved carried the same label of its parent follicle. Embryos were cultured. The embryologist was blinded. The oocyte and embryo quality were assessed and compared with the PFV of parent follicle. Results Follicles with grade 1 and 2 PFV were predominantly observed. The yield of oocytes was independent of PFV. The mean yield of good quality embryos in conjunction with the PFV of the parent follicle was found to be highly significant in both the groups. The antagonist group had statistically significant yield of mature oocytes and embryos, compared to the agonist group. Conclusion Antagonist protocol had favourable outcomes compared with the agonist protocol. The retrieval of oocytes, even the mature ones and the yield of high grade embryos were found higher. As the PFV increased, the yield and overall pregnancy rates were higher. PFV as assessed by Power Doppler is a useful non-invasive biomarker of embryo quality and can be employed in conjunction with other biomarkers in ART to predict successful outcome. PMID:26674932

Shared and Unique Patterns of Embryo Development in Extremophile Poeciliids

PubMed Central

Riesch, Rüdiger; Schlupp, Ingo; Langerhans, R. Brian; Plath, Martin

2011-01-01

Background Closely related lineages of livebearing fishes have independently adapted to two extreme environmental factors: toxic hydrogen sulphide (H2S) and perpetual darkness. Previous work has demonstrated in adult specimens that fish from these extreme habitats convergently evolved drastically increased head and offspring size, while cave fish are further characterized by reduced pigmentation and eye size. Here, we traced the development of these (and other) divergent traits in embryos of Poecilia mexicana from benign surface habitats (“surface mollies”) and a sulphidic cave (“cave mollies”), as well as in embryos of the sister taxon, Poecilia sulphuraria from a sulphidic surface spring (“sulphur mollies”). We asked at which points during development changes in the timing of the involved processes (i.e., heterochrony) would be detectible. Methods and Results Data were extracted from digital photographs taken of representative embryos for each stage of development and each type of molly. Embryo mass decreased in convergent fashion, but we found patterns of embryonic fat content and ovum/embryo diameter to be divergent among all three types of mollies. The intensity of yellow colouration of the yolk (a proxy for carotenoid content) was significantly lower in cave mollies throughout development. Moreover, while relative head size decreased through development in surface mollies, it increased in both types of extremophile mollies, and eye growth was arrested in mid-stage embryos of cave mollies but not in surface or sulphur mollies. Conclusion Our results clearly demonstrate that even among sister taxa convergence in phenotypic traits is not always achieved by the same processes during embryo development. Furthermore, teleost development is crucially dependent on sufficient carotenoid stores in the yolk, and so we discuss how the apparent ability of cave mollies to overcome this carotenoid-dependency may represent another potential mechanism explaining the lack of gene flow between surface and cave mollies. PMID:22087302

Observed Rate of Down Syndrome in Twin Pregnancies.

PubMed

Sparks, Teresa N; Norton, Mary E; Flessel, Monica; Goldman, Sara; Currier, Robert J

2016-11-01

To evaluate the observed incidence of Down syndrome in twins compared with that expected based on maternal age-matched singletons, which is the current clinical approach. This was a retrospective review of California Prenatal Screening Program participants with expected delivery dates between July 1995 and December 2012. Cases confirmed prenatally or postnatally with a genetic imbalance leading to phenotypic Down syndrome (trisomy 21, mosaic trisomy 21, or translocations) were included. Pregnancies conceived with ovum donation and women older than 45 years were excluded. We compared the observed Down syndrome incidence per pregnancy for twins with expected incidence by extrapolating from singleton data and expected zygosity as is the current clinical approach. This extrapolation assumes that monozygotic pregnancies have equivalent Down syndrome risk per pregnancy relative to maternal age-matched singletons and dizygotic pregnancies have twice the risk of at least one affected fetus. Zygosity for affected cases was presumed to be monozygotic with Down syndrome concordance and dizygotic with Down syndrome discordance. Counts were compared using cumulative Poisson distributions. Of 77,279 twin pregnancies, 182 (0.2%) had at least one fetus with Down syndrome confirmed by karyotype. The ratio of observed-to-expected Down syndrome incidence per pregnancy was 33.6%, 75.2%, and 70.0% for monozygotic, dizygotic, and all twins, respectively (P<.001 for all comparisons). Considering maternal age subgroups and twin zygosity, a significantly lower-than-expected Down syndrome incidence was seen for women aged 25 to 45 years with monozygotic pregnancies and overall for women aged 25 to 45 years with dizygotic pregnancies. The observed incidence of Down syndrome in twin pregnancies is lower than expected, most notably for monozygotic pregnancies and with increasing maternal age. Risk-based counseling can strongly affect women's choices regarding testing and management during pregnancy, so an understanding of the true Down syndrome risk in twin gestations is crucial.

Sperm-egg penetration assay assessment of the contraceptive effects of glycerol and egg yolk in rooster sperm diluents.

PubMed

Abouelezz, F M K; Castaño, C; Toledano-Díaz, A; Esteso, M C; López-Sebastián, A; Campo, J L; Santiago-Moreno, J

2015-06-01

Glycerol (GLY) and egg yolk (EY) are good cryoprotectants of avian and mammalian sperm, but in birds, they strongly inhibit the eventual fertilization of ova. Using the sperm penetration (SP-holes) assay and fertility trials, the present study investigates (1) the possible mechanism by which this contraceptive effect occurs in chickens and (2) the maximum concentrations of GLY and EY tolerated by fresh rooster sperm. Seventy Black-Barred Andaluza hens (five per treatment) were inseminated four times (twice per week) with 0.1 mL of fresh semen from roosters of the same breed diluted 1:1 (v:v) with Lake and Ravie medium containing different concentrations of GLY or EY. No adverse effects on acrosome integrity, sperm motility, or viability were seen with any concentration of GLY or EY. The number of SP-holes on perivitelline layer samples taken from above the germinal disc became progressively lower at GLY concentrations of 1.5% or greater (P > 0.05). No holes caused by sperms were seen in unfertilized eggs. The corresponding fertility results showed similar reductions when the GLY concentration was 1.5% or greater. No changes in the number of SP-holes were seen with increasing EY concentrations (0%-7.5%), nor were any differences in fertility observed, except for a reduction when 15% EY was used. The results therefore reveal that GLY affects the transit of sperms through the oviduct in their attempt to reach the infundibulum area, limiting their access to the ovum perivitelline layer. Egg yolk had no such effect, nor did it influence acrosome reaction capacity; its mechanism of contraceptive action therefore remains unknown. The maximum GLY and EY concentrations tolerated by the rooster sperm were 0.75% and 7.5%, respectively. Copyright © 2015 Elsevier Inc. All rights reserved.

An Electron Microscope Study of the Rat Ovum

PubMed Central

Sotelo, J. Roberto; Porter, Keith R.

1959-01-01

This paper reports on the fine structure of rat oocytes at stages before ovulation, during maturation, fertilization, and early cleavage. The study includes parallel observations on light and electron microscope preparations with attempted correlations. The follicular cells of the ovarian egg are described as sending long processes through the zona pellucida to the egg surface where they mingle with thin projections from the egg itself. No open communication between follicle cell cytoplasm and egg cytoplasm was observed. During maturation and fertilization both types of processes are withdrawn from the zona. The germinal vesicle and later the pronuclei of the fertilized egg are characterized by numerous large nucleoli. These have the form of thick walled vesicles with diameters as great as 8 to 10 µ. The wall is dense in the EM image and appears to consist in part of small granules. The cytoplasm shows several inclusions including mitochondria of usual form and a Golgi component which has the typical fine structure and the distribution described by earlier light studies. Small dense particles, presumably RNP particles, are distributed throughout the cytoplasmic matrix and show no preference for membranes. The endoplasmic reticulum of the oocyte is represented by a scattering only of vesicles, but begins a more extensive and elaborate development with the onset of segmentation. One inclusion of the ooplasm, similar in size to mitochondria, receives special attention. It is a vesicular structure, containing a large number of small vesicles (10 to 50 mµ in diameter) and frequently a central density or nucleoid. They are referred to as multivesicular bodies. Such bodies are found in small number in the ovarian egg, but increase greatly in number during maturation and fertilization. It appears from the micrographs of eggs in these latter stages that these vesicular bodies break down and liberate their content of small vesicles to the surrounding ooplasm. Comments are provided on the apparent significance of the various observations. PMID:13654454

Clinical and Analytical Evaluation of a Single-Vial Stool Collection Device with Formalin-Free Fixative for Improved Processing and Comprehensive Detection of Gastrointestinal Parasites

PubMed Central

Couturier, Brianne A.; Jensen, Ryan; Arias, Nora; Heffron, Michael; Gubler, Elyse; Case, Kristin; Gowans, Jason

2015-01-01

Microscopic examination of feces is a standard laboratory method for diagnosing gastrointestinal parasite infections. In North America, the ovum and parasite (O&P) examination is typically performed using stool that is chemically fixed in polyvinyl alcohol (PVA) and formalin, after which the stool is concentrated by filtration to enhance sensitivity. Mini Parasep solvent-free (SF) tubes allow collection and concentration within a single collection vial. The goal of the study was to determine whether consolidated processing and concentration with the Parasep tubes using an alcohol-based fixative (Alcorfix) provide O&P examinations equivalent to or better than those done by processing of PVA-formalin-fixed stool using a SpinCon concentration device. Parasep tubes revealed filtration performance equivalent to that of the SpinCon concentration device using PVA-formalin-fixed stool containing protozoa. Specimens cocollected in Parasep tubes containing PVA-formalin and Alcorfix revealed comparable morphology and staining for various protozoa. Alcorfix effectively fixed live Cryptosporidium and microsporidia such that morphology and staining were conserved for modified acid-fast and modified trichrome stains. A work flow analysis revealed significant time savings for batches of 10 or 30 O&P specimens in tubes with Alcorfix compared to the amount of time that it took to analyze the same number of specimens in tubes with PVA-formalin. The direct hands-on time savings with Mini Parasep tubes were 17 min and 41 s and 32 min and 1 s for batches of 10 and 30 specimens, respectively. Parasep tubes containing Alcorfix provide significant work flow advantages to laboratories that process medium to high volumes of O&P specimens by streamlining processing and converting to a single tube. These improvements in work flow, reduction of the amount of formalin used in the laboratory, and equivalent microscopy results are attractive advancements in O&P testing for North American diagnostic parasitology laboratories. PMID:26019199

Development of the follicle complex and oocyte staging in red drum, Sciaenops ocellatus Linnaeus, 1776 (Perciformes, Sciaenidae).

PubMed

Grier, Harry J

2012-08-01

Pelagic egg development in red drum, Sciaenops ocellatus, is described using tiered staging. Based on mitosis and meiosis, there are five periods: Mitosis of Oogonia, Active Meiosis I, Arrested Meiosis I, Active Meiosis II, and Arrested Meiosis II. The Periods are divided into six stages: Mitotic Division of Oogonia, Chromatin Nucleolus, Primary Growth, Secondary Growth, Oocyte Maturation and Ovulation. The Chromatin Nucleolus Stage is divided into four steps: Leptotene, Zygotene, Pachytene, and Early Diplotene. Oocytes in the last step possess one nucleolus, dispersed chromatin with forming lampbrush chromosomes and lack basophilic ooplasm. The Primary Growth Stage, characterized by basophilic ooplasm and absence of yolk in oocytes, is divided into five steps: One-Nucleolus, Multiple Nucleoli, Perinucleolar, Oil Droplets, and Cortical Alveolar. During primary growth, the Balbiani body develops from nuage, enlarges and disperses throughout the ooplasm as both endoplasmic reticulum and Golgi develop within it. Secondary growth or vitellogenesis has three steps: Early Secondary Growth, Late Secondary Growth and Full-Grown. The Oocyte Maturation Stage, including ooplasmic and germinal vesicle maturation, has four steps: Eccentric Germinal Vesicle, Germinal Vesicle Migration, Germinal Vesicle Breakdown and Resumption of Meiosis when complete yolk hydration occurs. The period is Arrested Meiosis II. When folliculogenesis is completed, the ovarian follicle, an oocyte and encompassing follicle cells, is surrounded by a basement membrane and developing theca, all forming a follicle complex. After ovulation, a newly defined postovulatory follicle complex remains attached to the germinal epithelium. It is composed of a basement membrane that separates the postovulatory follicle from the postovulatory theca. Arrested Meiosis I encompasses primary and secondary growth (vitellogenesis) and includes most of oocyte maturation until the resumption of meiosis (Active Meiosis II). The last stage, Ovulation, is the emergence of the oocyte from the follicle when it becomes an egg or ovum. Copyright © 2012 Wiley Periodicals, Inc.

[Prostaglandins in gynecology and obstetrics].

PubMed

Klausch, B; Kyank, H

1972-06-03

A review of early research (up through 1970) on prostaglandins (PGs) is presented. Their chemical structure and classification based on their ring-structure is detailed as well as various analytic methods of mammalian tissues and body fluids. For clinical use PGE1 and 2, PGF2alpha and PGA1 are the most significant ones because of their properties. PGs have many physiological activities encompassing many organ systems. Their pharmacological actions include: 1) stimulation of nonvascular smooth muscle; 2) peripheral vasodilation (excluding PGFs which cause vasoconstriction); 3) inhibition of lipolysis; 4) inhibition of platelet aggregation; 5) inhibition of gastric peristalsis and gastric juice secretion; 6) bronchodilation; and 7) inhibition of spontaneous CNS activity. The level of PGEs in semen is closely related to the degree of fertility; normally fertile men have 55 mcg PGE/ml and never less than 11 mcg/ml. Current studies are under way on the effect of PGE in artificial insemination of sperm of subfertile men. PGF2alpha and PGE2 stimulate menstruation and uterine contraction; other PGs inhibit uterine contraction. PGs from semen have a role in sperm transport and possibly act on fallopian tube motility aiding sperm capacitation, and ovum retention and transport. Early trials with PGs point to a possible action as an abortifacient, as a once-a-month contraceptive, or a postconception contraceptive agent. PGF2alpha is found in variable concentrations in maternal blood during contraction of the pregnant uterus; levels increase as labor progresses. PGs have been used for labor induction, for induction of abortion and in mole pregnancy. Given as a constant intravenous infusion they produce regular contractions leading to natural expulsion of the fetus and causing very few side effects in the woman with no adverse effects on the fetus. PGs' action compares favorably with that of oxytocin and is preferable for labor induction in certain pregnancy complications. PGE1 and 2 have a stronger effect than PGF2alpha, hence can be used in smaller dosage and cause fewer adverse effects.

Clinical and Analytical Evaluation of a Single-Vial Stool Collection Device with Formalin-Free Fixative for Improved Processing and Comprehensive Detection of Gastrointestinal Parasites.

PubMed

Couturier, Brianne A; Jensen, Ryan; Arias, Nora; Heffron, Michael; Gubler, Elyse; Case, Kristin; Gowans, Jason; Couturier, Marc Roger

2015-08-01

Microscopic examination of feces is a standard laboratory method for diagnosing gastrointestinal parasite infections. In North America, the ovum and parasite (O&P) examination is typically performed using stool that is chemically fixed in polyvinyl alcohol (PVA) and formalin, after which the stool is concentrated by filtration to enhance sensitivity. Mini Parasep solvent-free (SF) tubes allow collection and concentration within a single collection vial. The goal of the study was to determine whether consolidated processing and concentration with the Parasep tubes using an alcohol-based fixative (Alcorfix) provide O&P examinations equivalent to or better than those done by processing of PVA-formalin-fixed stool using a SpinCon concentration device. Parasep tubes revealed filtration performance equivalent to that of the SpinCon concentration device using PVA-formalin-fixed stool containing protozoa. Specimens cocollected in Parasep tubes containing PVA-formalin and Alcorfix revealed comparable morphology and staining for various protozoa. Alcorfix effectively fixed live Cryptosporidium and microsporidia such that morphology and staining were conserved for modified acid-fast and modified trichrome stains. A work flow analysis revealed significant time savings for batches of 10 or 30 O&P specimens in tubes with Alcorfix compared to the amount of time that it took to analyze the same number of specimens in tubes with PVA-formalin. The direct hands-on time savings with Mini Parasep tubes were 17 min and 41 s and 32 min and 1 s for batches of 10 and 30 specimens, respectively. Parasep tubes containing Alcorfix provide significant work flow advantages to laboratories that process medium to high volumes of O&P specimens by streamlining processing and converting to a single tube. These improvements in work flow, reduction of the amount of formalin used in the laboratory, and equivalent microscopy results are attractive advancements in O&P testing for North American diagnostic parasitology laboratories. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Developmental toxicology: adequacy of current methods.

PubMed

Peters, P W

1998-01-01

Toxicology embraces several disciplines such as carcinogenicity, mutagenicity and reproductive toxicity. Reproductive toxicology is concerned with possible effects of substances on the reproductive process, i.e. on sexual organs and their functions, endocrine regulation, fertilization, transport of the fertilized ovum, implantation, and embryonic, fetal and postnatal development, until the end-differentiation of the organs is achieved. Reproductive toxicology is divided into areas related to male and female fertility, and developmental toxicology. Developmental toxicology can be further broken down into prenatal and postnatal toxicology. Today, much new information is available about the origins of developmental disorders resulting from chemical exposure. While these findings seem to promise important new developments in methodology and research, there is a danger of losing sight of the precepts and principles established in the light of existing knowledge. There is also a danger that we may fail to correct shortcomings in our existing procedures and practice. The aim of this presentation is to emphasize the importance of testing substances for their impact in advance of their use and to underline that we must use the best existing tools for carrying out risk assessments. Moreover, it needs to be stressed that there are many substances that are never assessed with respect to reproductive and developmental toxicity. Similarly, our programmes for post-marketing surveillance with respect to developmental toxicology are grossly inadequate. Our ability to identify risks to normal development and reproduction would be much improved, first if a number of straightforward precepts were always followed and second, if we had a clearer understanding of what we mean by risk and acceptable levels of risk in the context of development. Other aims of this paper are: to stress the complexity of the different stages of normal prenatal development; to note the principles that are applicable in developmental and especially prenatal toxicology; to describe the different agents that might act as developmental toxicants or teratogens; to show the broad scope of different effects caused by developmental toxic agents; and to indicate methods to detect and to recognise causes of developmental defects with the primary objective of preventing these disorders.

Early in vitro fertilization improves development of bovine ova heat stressed during in vitro maturation.

PubMed

Schrock, G E; Saxton, A M; Schrick, F N; Edwards, J L

2007-09-01

The objectives were to examine the development of embryos derived from control (38.5 degrees C) or heat-stressed ova [41.0 degrees C during the first 12 h of in vitro maturation (hIVM)] when in vitro fertilization (IVF) was performed at 16, 18, 20, 24, or 30 hIVM. Effects of heat stress in compromising ovum development depended on when IVF was performed (in vitro maturation temperature x IVF time interaction). When IVF was performed at 24 or 30 hIVM, fewer heat-stressed ova developed to the blastocyst stage compared with the respective controls. In contrast, when IVF was performed at 16, 18, or 20 hIVM, more heat-stressed ova developed to the blastocyst stage compared with the respective controls. Performing IVF earlier than usual was beneficial, because the ability of heat-stressed ova to develop to the blastocyst stage was improved when IVF was performed at 18 or 20 vs. 24 hIVM. Blastocyst stage and quality were equivalent to non-heat-stressed controls regardless of IVF time. Control ova undergoing IVF at 20, 24, 30, or 32 hIVM and heat-stressed ova undergoing IVF at 16, 18, 20, or 24 hIVM were compared for blastocyst development by multisource regression. Although linear and quadratic slopes were similar, heat stress reduced the peak and shifted the developmental response of ova by 7.3 h. In other words, obtaining optimal blastocyst development from heat-stressed ova would depend on performing IVF at 19.5 hIVM compared with 26.7 hIVM for non-heat-stressed controls. Heat-induced reductions in peak blastocyst development significantly reduced the window of time available to perform IVF and obtain > or = 20% blastocyst development. In summary, results support an effect of heat stress to hasten developmentally important events during oocyte maturation. The inability of earlier IVF to fully restore the development of heat-stressed ova to that of non-heat-stressed controls highlights the importance of further study.

Physiological differences beween fertilized and unfertilized mouse ova: glycerol permeability and freezing sensitivity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jackowski, S.C.

The glycerol permeability and freezing sensitivity of mouse ova were studied for evidence of differences associated with fertilization. The times of ovulation, fertilization and first cleavage of ova were determined as a function of time after the administration of human chorionic gonadotropic, hormone, HCG, to female mice. Fertilization did not cause a large instantaneous change in glycerol permeability. Rather, the permeability coefficient for glycerol at approximately 3/sup 0/C gradually increased from 7.0 x 10/sup -7/ to 7.0 x 10/sup -6/ cm/min for fertilized ova isolated from about 1 hour to 16 hours after fertilization. The zonae pellucidae of fertilized andmore » unfertilized ova did not act as detectable barriers to permeation by glycerol. No significant and immediate change was observed on the surface of the ovum as a result of fertilization. Survival after freezing was assayed by two techniques: measurement of the ability of the cells to fluoresce in the presence of fluorescein diacetate; successful development in culture. Survival of fertilized and unfertilized ova increased as a function of both the temperature and time of incubation in glycerol prior to freezing. It was concluded that permeation of a cell by glycerol enhances survival. The cooling rate that yielded optimal survival of zygotes in G/sub 2/ phase differed from that of unfertilized ova and zygotes in G/sub 1/ phase. The optimum rate for the latter cells was about 1/sup 0/C/min with survival being about 63 percent and 79 percent, respectively. The optimum rate for zygotes in G/sub 2/ ranged from 1/sup 0/C/min to 7/sup 0/C/min with survival being about 58 percent. The differences among the freezing sensitivities of unfertilized ova, zygotes in G/sub 1/ and zygotes in G/sub 2/ can be explained in terms of their differences in glycerol permeability and possibly in terms of the increased surface area associated with the number of microvilli on the G/sub 2/ zygotes.« less

Estimating variance components and breeding values for number of oocytes and number of embryos in dairy cattle using a single-step genomic evaluation.

PubMed

Cornelissen, M A M C; Mullaart, E; Van der Linde, C; Mulder, H A

2017-06-01

Reproductive technologies such as multiple ovulation and embryo transfer (MOET) and ovum pick-up (OPU) accelerate genetic improvement in dairy breeding schemes. To enhance the efficiency of embryo production, breeding values for traits such as number of oocytes (NoO) and number of MOET embryos (NoM) can help in selection of donors with high MOET or OPU efficiency. The aim of this study was therefore to estimate variance components and (genomic) breeding values for NoO and NoM based on Dutch Holstein data. Furthermore, a 10-fold cross-validation was carried out to assess the accuracy of pedigree and genomic breeding values for NoO and NoM. For NoO, 40,734 OPU sessions between 1993 and 2015 were analyzed. These OPU sessions originated from 2,543 donors, from which 1,144 were genotyped. For NoM, 35,695 sessions between 1994 and 2015 were analyzed. These MOET sessions originated from 13,868 donors, from which 3,716 were genotyped. Analyses were done using only pedigree information and using a single-step genomic BLUP (ssGBLUP) approach combining genomic information and pedigree information. Heritabilities were very similar based on pedigree information or based on ssGBLUP [i.e., 0.32 (standard error = 0.03) for NoO and 0.21 (standard error = 0.01) for NoM with pedigree, 0.31 (standard error = 0.03) for NoO, and 0.22 (standard error = 0.01) for NoM with ssGBLUP]. For animals without their own information as mimicked in the cross-validation, the accuracy of pedigree-based breeding values was 0.46 for NoO and NoM. The accuracies of genomic breeding values from ssGBLUP were 0.54 for NoO and 0.52 for NoM. These results show that including genomic information increases the accuracies. These moderate accuracies in combination with a large genetic variance show good opportunities for selection of potential bull dams. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Natural family planning.

PubMed

Bourdillon, C

1982-11-01

Frequently, when one mentions natural family planning methods, the response is doubt, bewilderment, ridicule, or scorn. Much of this is due to the fact that many people know only of the rhythm method, which depends on a calculation based on the menstrual pattern. What many people do not know, including physicians and nurses, is that in Australia Drs. John and Lyn Billings have been scientifically researching natural methods of family planning for 25 years, and they have pioneered this Ovulation Method of family planning. The Billings Ovulation Method depends only on the mucus sign. It is based on a scientific knowledge of the combined fertility of husband and wife, and an understanding of the physiology of the female body through simple observations. Important facts relating to the practice of the method include: ovulation takes place only once in the cycle; mucus is secreted by the cervical mucosa before ovulation; the ovum lives for only 3 days at the most; and the sperms live for 5 days at the most, and only in the presence of this fertile mucus. There a 2 types of mucus. The 1st type to appear is cloudy or white. It is nonslippery, sticky, and breaks when stretched between 2 fingers. The 2nd type is like the part of a hen's egg. It is very slippery, much clearer than the former, and stretches when pulled between 2 fingers. This is fertile mucus, and ovulation occurs on the last day that this is present. Of course, it cannot be recognized as the last day until its absence is observed on the following day. The mucus sign can be both seen and felt. After a few months a woman will readily recognize her fertile time, but daily charting is advocated. Simple signs, representing the various observations, are taught. Once a couple knows and understands their combined fertility through the observation of the mucus sign they can plan their family. Rules of the method are outlined. The self control required by this method can only serve to increase the selfless love and unselfish care and respect between partners.

The low fertility of repeat-breeder cows during summer heat stress is related to a low oocyte competence to develop into blastocysts.

PubMed

Ferreira, R M; Ayres, H; Chiaratti, M R; Ferraz, M L; Araújo, A B; Rodrigues, C A; Watanabe, Y F; Vireque, A A; Joaquim, D C; Smith, L C; Meirelles, F V; Baruselli, P S

2011-05-01

It was hypothesized the lower fertility of repeat-breeder (RB) Holstein cows is associated with oocyte quality and this negative effect is enhanced during summer heat stress (HS). During the summer and the winter, heifers (H; n=36 and 34, respectively), peak-lactation (PL; n=37 and 32, respectively), and RB (n=36 and 31, respectively) Holstein cows were subjected to ovum retrieval to assess oocyte recovery, in vitro embryonic developmental rates, and blastocyst quality [terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and total cell number]. The environmental temperature and humidity, respiration rate, and cutaneous and rectal temperatures were recorded in both seasons. The summer HS increased the respiration rate and the rectal temperature of PL and RB cows, and increased the cutaneous temperature and lowered the in vitro embryo production of Holstein cows and heifers. Although cleavage rate was similar among groups [H=51.7% ± 4.5 (n=375), PL=37.9% ± 5.1 (n=390), RB=41.9% ± 4.5 (n=666)], blastocyst rate was compromised by HS, especially in RB cows [H=30.3% ± 4.8 (n=244) vs. 23.3% ± 6.4 (n=150), PL=22.0% ± 4.7 (n=191) vs. 14.6% ± 7.6 (n=103), RB=22.5% ± 5.4 (n=413) vs. 7.9% ± 4.3 (n=177)]. Moreover, the fragmentation rate of RB blastocysts was enhanced during the summer, compared with winter [4.9% ± 0.7 (n=14) vs. 2.2% ± 0.2 (n=78)] and other groups [H=2.5% ± 0.7 (n=13), and PL=2.7% ± 0.6 (n=14)] suggesting that the association of RB fertility problems and summer HS may potentially impair oocyte quality. Our findings provide evidence of a greater sensitivity of RB oocytes to summer HS. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The link between oral contraceptive use and prevalence in autism spectrum disorder.

PubMed

Strifert, Kim

2014-12-01

Autism spectrum disorder (ASD) is a group of developmental disabilities that include full syndrome autism, Asperger's syndrome, and other pervasive developmental disorders. The identified prevalence of ASD has increased in a short time period across multiple studies causing some to conclude that it has reached epidemic proportions in the U.S. Many possible explanations for the rise in numbers of individuals diagnosed with ASD have been offered and yet, causes and contributing factors for ASD are inadequately understood. Current evidence suggests that both genetics and environment play a part in causing ASD. One possible risk factor for the increase in prevalence has been profoundly overlooked in the existing biomedical and epidemiologic literature. As the prevalence of ASD has risen in the last sixty years, so has the prevalence of the usage of the oral contraceptives and other modern hormonal delivery methods. In 1960 about one million American women were using oral contraceptives, today close to 11 million women in the U.S. use oral contraceptives. Eighty-two percent of sexually active women in the U.S. have used oral contraceptives at some point during their reproductive years. Thus, the growth in use of progesterone/estrogen-based contraceptives in the United State has reached near-ubiquitous levels among women in the child-bearing age range. The suppression of ovulation produced by estrogen-progesterone is an indisputable abnormality. It is logical to consider the outcome of the ovum that would have been normally released from the ovary during ovulation. To date there is no comprehensive research into the potential neurodevelopmental effects of oral contraceptive use on progeny. The issue has been only sparsely considered in the biomedical literature. This article hypothesizes that the compounds, estrogen and progesterone, used in oral contraceptives modify the condition of the oocyte and give rise to a potent risk factor that helps explain the recent increase in the prevalence of ASD's. This hypothesis does not propose to delineate the cause of autism. Rather, it attempts to explain the recent dramatic increase in prevalence and point the way for further study that will lead to causal examination. Copyright © 2014 The Author. Published by Elsevier Ltd.. All rights reserved.

Prenatal and postnatal maternal contributions to reproductive, maternal, and size-related traits of beef cattle.

PubMed

Gregory, K E; Maurer, R R

1991-03-01

Brown Swiss-Hereford (BS-H) reciprocal cross embryos were transferred to BS and H recipient cows and Red Poll-Angus (RP-A) reciprocal cross embryos were transferred to RP and A recipient cows to estimate the relative contributions of ovum cytoplasm and uterine influences to prenatal maternal effects. Calves resulting from embryo transfers (ET) were weaned early (3 to 5 d). Reciprocal cross mating also were made by natural service (NS) between BS and H and between RP and A breeds; part of the offspring were weaned at 3 to 5 d, and the remainder nursed their dams to an age of 150 to 180 d. This was done to estimate breed differences in prenatal and postnatal effects combined and to separate the effects of prenatal maternal influences from postnatal maternal influences of these breeds. Females produced in both ET and NS parts of the experiment were retained to produce three calf crops to an age of about 4.5 yr. The following traits were analyzed: percentage of conception rate; percentage of calf survival; percentage of calves produced per cow exposed; birth and weaning weights of calves produced; and periodic weights, heights, and condition scores of females to an age of 4.5 yr. Neither breed of donor (cytoplasmic influence) nor breed of recipient (uterine influence) had consistently important effects on the traits evaluated. In NS matings, differences between reciprocal crosses were small for most of the traits evaluated. Method of rearing (nursed vs weaned at 3 to 5 d) had no effect on reproductive and maternal traits for RP-A reciprocal cross females, but females that nursed generally were heavier, were taller, and had higher condition scores at most ages than early-weaned females. For the BS-H reciprocal cross, early-weaned females were favored over females reared by their dams in percentage of calves produced per cow exposed, but the method of rearing did not affect other reproductive or maternal traits. BS-H reciprocal cross females that nursed their dams were heavier at 550 d and were heavier and had higher condition scores at an age of 34 mo than early-weaned females.

[Correlation of the DNA fragmentation index and malformation rate of optimized sperm with embryonic development and early spontaneous abortion in IVF-ET].

PubMed

Jiang, Wei-Jie; Jin, Fan; Zhou, Li-Ming

2016-06-01

To investigate the effects of the DNA fragmentation index (DFI) and malformation rate (SMR) of optimized sperm on embryonic development and early spontaneous abortion in conventional in vitro fertilization and embryo transfer (IVF-ET). We selected 602 cycles of conventional IVF-ET for pure oviductal infertility that had achieved clinical pregnancies, including 505 cycles with ongoing pregnancy and 97 cycles with early spontaneous abortion. On the day of ovum retrieval, we examined the DNA integrity and morphology of the rest of the optimized sperm using the SCD and Diff-Quik methods, established the joint predictor (JP) by logistic equation, and assessed the value of DFI and SMR in predicting early spontaneous abortion using the ROC curve. The DFI, SMR, and high-quality embryo rate were (15.91±3.69)%, (82.85±10.24)%, and 46.53% (342/735) in the early spontaneous abortion group and (9.30±4.22)%, (77.32±9.19)%, and 56.43% (2263/4010) respectively in the ongoing pregnancy group, all with statistically significant differences between the two groups (P<0.05 ). Both the DFI and SMR were the risk factors of early spontaneous abortion (OR = 5.96 and 1.66; both P< 0.01). The areas under the ROC curve for DFI, SMR and JP were 0.893±0.019, 0.685±0.028, and 0.898±0.018, respectively. According to the Youden index, the optimal cut-off values of the DFI and SMR obtained for the prediction of early spontaneous abortion were approximately 15% and 80%. The DFI was correlated positively with SMR (r= 0.31, P<0.01) but the high-quality embryo rate negatively with both the DFI (r= －0.45, P<0.01) and SMR (r= －0.22, P<0.01). The DFI and SMR of optimized sperm are closely associated with embryonic development in IVF. The DFI has a certain value for predicting early spontaneous abortion with a threshold of approximately 15%, but SMR may have a lower predictive value.

Myo-inositol may improve oocyte quality in intracytoplasmic sperm injection cycles. A prospective, controlled, randomized trial.

PubMed

Papaleo, Enrico; Unfer, Vittorio; Baillargeon, Jean-Patrice; Fusi, Francesco; Occhi, Francesca; De Santis, Lucia

2009-05-01

To determine the effects of myo-inositol on oocyte quality in polycystic ovary syndrome (PCOS) patients undergoing intracytoplasmic sperm injection (ICSI) cycles. A prospective, controlled, randomized trial. Assisted reproduction centers. Sixty infertile PCO patients undergoing ovulation induction for ICSI. All participants underwent standard long protocol. Starting on the day of GnRH administration, 30 participants received myo-inositol combined with folic acid (Inofolic) 2 g twice a day and 30 control women received folic acid alone, administrated continuously. Primary end points were number of morphologically mature oocytes retrieved, embryo quality, and pregnancy and implantation rates. Secondary end points were total number of days of FSH stimulation, total dose of gonadotropin administered, E(2) level on the day of hCG administration, fertilization rate per number of retrieved oocytes, embryo cleavage rate, live birth and miscarriage rates, cancellation rate, and incidence of moderate or severe ovarian hyperstimulation syndrome. Total r-FSH units (1,958 +/- 695 vs. 2,383 +/- 578) and number of days of stimulation (11.4 +/- 0.9 vs. 12.4 +/- 1.4) were significantly reduced in the myo-inositol group. Furthermore, peak E(2) levels (2,232 +/- 510 vs. 2,713 +/- 595 pg/mL) at hCG administration were significantly lower in patients receiving myo-inositol. The mean number of oocytes retrieved did not differ in the two groups, whereas in the group cotreated with myo-inositol the mean number of germinal vesicles and degenerated oocytes was significantly reduced (1.0 +/- 0.9 vs. 1.6 +/- 1.0), with a trend for increased percentage of oocytes in metaphase II (0.82 +/- 0.11% vs. 0.75 +/- 0.15%). These data show that in patients with PCOS, treatment with myo-inositol and folic acid, but not folic acid alone, reduces germinal vesicles and degenerated oocytes at ovum pick-up without compromising total number of retrieved oocytes. This approach, reducing E(2) levels at hGC administration, could be adopted to decrease the risk of hyperstimulation in such patients.

The evolution of health policy guidelines for assisted reproduction in the Republic of Ireland, 2004-2009.

PubMed

Walsh, David J; Ma, Mary L; Sills, Eric Scott

2011-06-24

This analysis reports on Irish regulatory policies for in vitro fertilisation (IVF) from 2004-2009, in the context of membership changes within the Medical Council of Ireland. To achieve this, the current (2009) edition of the Guide to Professional Conduct & Ethics was compared with the immediately preceding version (2004). The statutory composition of the Medical Council from 2004-2009 was also studied. Content analysis of the two editions identified the following differences: 1) The 2004 guide states that IVF "should only be used after thorough investigation has failed to reveal a treatable cause of the infertility", while the 2009 guide indicates IVF "should only be used after thorough investigation has shown that no other treatment is likely to be effective"; 2) The 2004 stipulation stating that fertilized ovum (embryo) "must be used for normal implantation and must not be deliberately destroyed" is absent from the 2009 guidelines; 3) The option to donate "unused fertilised ova" (embryos) is omitted from the 2009 guidelines; 4) The 2009 guidelines state that ART should be offered only by "suitably qualified professionals, in appropriate facilities, and according to the international best practice"; 5) The 2009 guidelines introduce criteria that donations as part of a donor programme should be "altruistic and non-commercial". These last two points represent original regulatory efforts not appearing in the 2004 edition. The Medical Practitioners Act 2007 reduced the number of physicians on the Medical Council to 6 (of 25) members. The ethical guidelines from 2004 preceded this change, while the reconstituted Medical Council published the 2009 version. Between 2004 and 2009, substantial modifications in reproductive health policy were incorporated into the Medical Council's ethical guidelines. The absence of controlling Irish legislation means that patients and IVF providers in Ireland must rely upon these guidelines by default. Our critique traces the evolution of public policy on IVF during a time when the membership of the Medical Council changed radically; reduced physician contribution to decision-making was associated with diminished protection for IVF-derived embryos in Ireland. Considerable uncertainty on IVF practice in Ireland remains.

Schoolchildren, maternal nutrition and generating healthy brains: the importance of lifecycle education for fertility, health and peace.

PubMed

House, Simon H

2009-01-01

In order to generate healthy brains, the earlier in life that children develop a healthy lifestyle the better. At home and at school, with a good diet and physical activity, children need to experience and to understand the value of healthy metabolism. Continuous education is needed in lifecycle health, with awareness that the most vulnerable phase relates to reproduction. This is the way that most people can become parents of healthy children. Nutrient deficits, toxins, or uro-genital disease, often unnoticed, affect quality of sperm or ovum, and subsequent development. Early errors can have the most pronounced effect. Gene-switches are being set early in life. Any early flaw can be magnified by growth. Moreover children's habits become set and young couples are too busy to learn and adapt. Many are liable to unintended pregnancy. Adoption of a healthy lifestyle at a young age is the most reliable route to producing healthy babies. The mother's state throughout pregnancy, emotions/hormones included, naturally continues to powerfully affect her child's development, therefore future physical and mental health, behaviour and ability. There has recently been a serious increase in babies conceived by schoolchildren, as well as a shift by working women towards childbearing in their late thirties or more. A return towards childbearing in the twenties and early thirties can reduce risks for the child. The mother needs to build her appropriate body-stores--vitamins and minerals, proteins, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and other essential polyunsaturated fatty acids (PUFA)--to ensure optimal development of her child. Replenishment of reserves is important for maternal health and before bearing another child. A restorative time of three years is desirable. Governments and school teachers need to guide and encourage parents in this health education and practice, and to use their authority to achieve it in schools. Empowerment with knowledge is the one way to fend off the growing pandemic of mental ill health and related disorders, and to make the most of a nation's genetic potential. There could be no better investment financially, let alone in enhancing people's lives.

Effects of Different Maturation Systems on Bovine Oocyte Quality, Plasma Membrane Phospholipid Composition and Resistance to Vitrification and Warming.

PubMed

Sprícigo, José F W; Diógenes, Mateus N; Leme, Ligiane O; Guimarães, Ana L; Muterlle, Carolle V; Silva, Bianca Damiani Marques; Solà-Oriol, David; Pivato, Ivo; Silva, Luciano Paulino; Dode, Margot A N

2015-01-01

The objective of this study was to evaluate the effects of different maturation systems on oocyte resistance after vitrification and on the phospholipid profile of the oocyte plasma membrane (PM). Four different maturation systems were tested: 1) in vitro maturation using immature oocytes aspirated from slaughterhouse ovaries (CONT; n = 136); 2) in vitro maturation using immature oocytes obtained by ovum pick-up (OPU) from unstimulated heifers (IMA; n = 433); 3) in vitro maturation using immature oocytes obtained by OPU from stimulated heifers (FSH; n = 444); and 4) in vivo maturation using oocytes obtained from heifers stimulated 24 hours prior by an injection of GnRH (MII; n = 658). A sample of matured oocytes from each fresh group was analyzed by matrix associated laser desorption-ionization (MALDI-TOF) to determine their PM composition. Then, half of the matured oocytes from each group were vitrified/warmed (CONT VIT, IMA VIT, FSH VIT and MII VIT), while the other half were used as fresh controls. Afterwards, the eight groups underwent IVF and IVC, and blastocyst development was assessed at D2, D7 and D8. A chi-square test was used to compare embryo development between the groups. Corresponding phospholipid ion intensity was expressed in arbitrary units, and following principal components analyses (PCA) the data were distributed on a 3D graph. Oocytes obtained from superstimulated animals showed a greater rate of developmental (P<0.05) at D7 (MII = 62.4±17.5% and FSH = 58.8±16.1%) compared to those obtained from unstimulated animals (CONT = 37.9±8.5% and IMA = 50.6±14.4%). However, the maturation system did not affect the resistance of oocytes to vitrification because the blastocyst rate at D7 was similar (P>0.05) for all groups (CONT VIT = 2.8±3.5%, IMA VIT = 2.9±4.0%, FSH VIT = 4.3±7.2% and MII VIT = 3.6±7.2%). MALDI-TOF revealed that oocytes from all maturation groups had similar phospholipid contents, except for 760.6 ([PC (34:1) + H]+), which was more highly expressed in MII compared to FSH (P<0.05). The results suggest that although maturation systems improve embryonic development, they do not change the PM composition nor the resistance of bovine oocytes to vitrification.

Effects of Different Maturation Systems on Bovine Oocyte Quality, Plasma Membrane Phospholipid Composition and Resistance to Vitrification and Warming

PubMed Central

Sprícigo, José F. W.; Diógenes, Mateus N.; Leme, Ligiane O.; Guimarães, Ana L.; Muterlle, Carolle V.; Silva, Bianca Damiani Marques; Solà-Oriol, David; Pivato, Ivo; Silva, Luciano Paulino; Dode, Margot A. N.

2015-01-01

The objective of this study was to evaluate the effects of different maturation systems on oocyte resistance after vitrification and on the phospholipid profile of the oocyte plasma membrane (PM). Four different maturation systems were tested: 1) in vitro maturation using immature oocytes aspirated from slaughterhouse ovaries (CONT; n = 136); 2) in vitro maturation using immature oocytes obtained by ovum pick-up (OPU) from unstimulated heifers (IMA; n = 433); 3) in vitro maturation using immature oocytes obtained by OPU from stimulated heifers (FSH; n = 444); and 4) in vivo maturation using oocytes obtained from heifers stimulated 24 hours prior by an injection of GnRH (MII; n = 658). A sample of matured oocytes from each fresh group was analyzed by matrix associated laser desorption-ionization (MALDI-TOF) to determine their PM composition. Then, half of the matured oocytes from each group were vitrified/warmed (CONT VIT, IMA VIT, FSH VIT and MII VIT), while the other half were used as fresh controls. Afterwards, the eight groups underwent IVF and IVC, and blastocyst development was assessed at D2, D7 and D8. A chi-square test was used to compare embryo development between the groups. Corresponding phospholipid ion intensity was expressed in arbitrary units, and following principal components analyses (PCA) the data were distributed on a 3D graph. Oocytes obtained from superstimulated animals showed a greater rate of developmental (P<0.05) at D7 (MII = 62.4±17.5% and FSH = 58.8±16.1%) compared to those obtained from unstimulated animals (CONT = 37.9±8.5% and IMA = 50.6±14.4%). However, the maturation system did not affect the resistance of oocytes to vitrification because the blastocyst rate at D7 was similar (P>0.05) for all groups (CONT VIT = 2.8±3.5%, IMA VIT = 2.9±4.0%, FSH VIT = 4.3±7.2% and MII VIT = 3.6±7.2%). MALDI-TOF revealed that oocytes from all maturation groups had similar phospholipid contents, except for 760.6 ([PC (34:1) + H]+), which was more highly expressed in MII compared to FSH (P<0.05). The results suggest that although maturation systems improve embryonic development, they do not change the PM composition nor the resistance of bovine oocytes to vitrification. PMID:26107169

Short communication: Genetic correlations between number of embryos produced using in vivo and in vitro techniques in heifer and cow donors.

PubMed

Jaton, C; Koeck, A; Sargolzaei, M; Price, C A; Baes, C; Schenkel, F S; Miglior, F

2016-10-01

Multiple embryos can be produced from a heifer or cow donors using an in vivo or an in vitro technique. Comparisons of the number of embryos produced by the same donors as heifers and cows and using different techniques are limited. The main objectives of this study were to assess the genetic correlation between the number of embryos produced by Holstein donors using an in vivo and in vitro technique as a heifer and as a cow. The data set used was recorded by Holstein Canada and included all successful superovulations or ovum pickup and in vitro fertilization procedures performed on Holstein donors for more than 20yr. The type of technique used was known for all records and the status of the donor at recovery was retrieved from calving records. Bivariate repeatability animal model analyses were performed for both the total number of embryos (NE) and the number of viable embryos (VE) recovered per procedure. Logarithmic transformation was performed on the traits to normalize the data. Heritability estimates for the donor varied between 0.14 (0.02) and 0.19 (0.03) over all analyses, indicating that the number of embryos produced by a donor is influenced by the genetic potential of the donor. Genetic correlations between records produced in vivo and in vitro were moderately high and positive (NE=0.85±0.07; VE=0.63±0.09), suggesting that donors with high genetic potential for in vivo superovulation tend also to have high potential to produce multiple embryos in vitro. Similarly, the moderately high genetic correlations (NE=0.79±0.05; VE=0.72±0.05) found between heifer and cow records indicate that a donor tends to produce a comparable number of embryos as a heifer or as a cow. The estimated repeatabilities (0.23 to 0.35) indicated that the number of embryos recovered should be somewhat repeatable in the same donor over time. On the other hand, the service sires seem not to play an important role on the total number of embryos produced by a donor no matter the technique used or the status of the donor at recovery. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Advances in human reproductive ecology.

PubMed

Ellison, P T

1994-01-01

Human reproductive ecology pertains to reproduction biology and changes due to environmental influences. The research literature relies on clinical, epidemiological, and demographic analysis. The emphasis is on normal, nonpathological states and a broad range of ecological conditions. This review focused on the importance of age and energetic stress from ecological conditions rather than dieting or self-directed exercise in changing female fecundity. The literature on male reproductive ecology is still small but growing. J.W. Wood provided a comprehensive overview of the field. Natural fertility, as defined by Henry, is the lack of parity-specific fertility limitation. There is evidence that fertility can vary widely in natural fertility populations. There are consistent age patterns among different natural fertility populations. Doring found that there was higher frequency of anovulatory and luteal insufficiency in cycles during perimenarche and perimenopausal periods. Infertility studies have shown declines in pregnancy rates in women over the age of 30 years. Ovum donation evaluations have found both uterine age and ovarian and oocyte age to be related to the probability of a successful pregnancy. Basal follicle stimulating hormone and the endometrial thickness are important predictors of ovarian capacity and related to age and declining fecundity. Much of the literature on fecundity is derived from women with impaired reproductive physiology. In Lipson and Ellison's study of healthy women, average follicular and average luteal estradiol values declined with increasing subject age. Low follicular levels were correlated with smaller follicular size, low oocyte fertilizability, reduced endometrial thickness, and low pregnancy rates. Comparisons across populations have shown that populations experience declines in luteal function with age, but levels of luteal functions varied widely. Chronic conditions which slow growth and delay reproductive maturation may impact on lower ovarian function throughout adult life. There is a range of ovarian function along a continuum due to energetic stress. Evidence from the Lese in Zaire, the Tamang of Nepal, and Polish farm women outside Crakow suggest that workload affects ovarian function. Luteal function and ovulatory frequency is lower when women are losing weight. Among the Tamang losing weight between seasons there was evidence of lower ovarian function during the monsoon season. Polish farm women who work very hard in summer had lower ovarian function. The effect of lactation on amenorrhea appears to be due to the energetic stress on the mother in the intensity and duration of suckling. Women in poorer nutritional status may require more intense suckling. Seasonality of energy balance may be related to seasonality of female fecundity and conceptions.

Effects of concentrate type and chromium propionate on insulin sensitivity, productive and reproductive parameters of lactating dairy cows consuming excessive energy.

PubMed

Leiva, T; Cooke, R F; Brandão, A P; Pardelli, U; Rodrigues, R O; Corrá, F N; Vasconcelos, J L M

2017-03-01

This experiment compared insulin sensitivity parameters, milk production and reproductive outcomes in lactating dairy cows consuming excessive energy, and receiving in a 2×2 factorial arrangement design: (1) concentrate based on ground corn (CRN; n=13) or citrus pulp (PLP; n=13), and (2) supplemented (n=14) or not (n=12) with 2.5 g/day of chromium (Cr)-propionate. During the experiment (day 0 to 182), 26 multiparous, non-pregnant, lactating Gir×Holstein cows (initial days in milk=80±2) were offered corn silage for ad libitum consumption, and individually received concentrate formulated to allow diets to provide 160% of their daily requirements of net energy for lactation. Cow BW and body condition score (BCS) were recorded weekly. Milk production was recorded daily and milk samples collected weekly. Blood samples were collected weekly before the morning concentrate feeding. Glucose tolerance tests (GTT; 0.5 g of glucose/kg of BW) were performed on days -3, 60, 120 and 180. Follicle aspiration for in vitro embryo production was performed via transvaginal ovum pick-up on days -1, 82 and 162. No treatment differences were detected (P⩾0.25) for BW and BCS change during the experiment. Within weekly blood samples, concentrations of serum insulin and glucose, as well as insulin : glucose ratio were similar among treatments (P⩾0.19), whereas CRN had less (P<0.01) non-esterified fatty acid concentrations compared with PLP (0.177 v. 0.215 mmol/l; SEM=0.009). During the GTT, no treatment differences were detected (P⩾0.16) for serum glucose concentration, glucose clearance rate, glucose half-life and insulin : glucose ratio. Serum insulin concentrations were less (P=0.04) in CRN supplemented with Cr-propionate compared with non-supplemented CRN (8.2 v. 13.5 µIU/ml, respectively; SEM=1.7), whereas Cr-propionate supplementation did not impact (P=0.70) serum insulin within PLP cows. Milk production, milk fat and solid concentrations were similar (P⩾0.48) between treatments. However, CRN had greater (P<0.01) milk protein concentration compared with PLP (3.54% v. 3.14%, respectively; SEM=0.08). No treatment differences were detected (P⩾0.35) on number of viable oocytes collected and embryos produced within each aspiration. In summary, feeding a citrus pulp-based concentrate to lactating dairy cows consuming excessive energy did not improve insulin sensitivity, milk production and reproductive outcomes, whereas Cr-propionate supplementation only enhanced insulin sensitivity in cows receiving a corn-based concentrate during a GTT.

Injection-Site Reactions in Wild Horses (Equus caballus) Receiving an Immunocontraceptive Vaccine

USGS Publications Warehouse

Roelle, James E.; Ransom, Jason I.

2009-01-01

The U.S. Geological Survey and the Bureau of Land Management are conducting research on the efficacy of the immunocontraceptive agent porcine zona pellucida (PZP) in reducing fertility of wild horses (Equus caballus). As an antigen, PZP stimulates antibody production when injected into many mammalian species. These antibodies bind to the external surface of the ovum, preventing fertilization. By itself, PZP is only weakly immunogenic and is therefore delivered with an adjuvant, most commonly one of the Freund adjuvants, designed to further stimulate antibody production. Freund's complete adjuvant (FCA) in particular is known to be very effective, but may also be associated with undesirable side effects such as formation of abscesses at injection sites. Such reactions may be exacerbated when accompanied by the additional trauma of a remotely delivered dart. Because horses in our three study herds were individually identifiable by color markings and harem association, we were able to monitor mares for injection-site reactions (abscesses, nodules, swelling, and stiffness) following inoculation with PZP. In 100 injections delivered by hand we observed a single nodule, two instances of swelling, and no other reactions. In two herds that received remotely delivered (dart) injections, the frequency of reactions was about 1 and 6 percent for abscesses, 25 percent for nodules (both herds), 11 and 33 percent for swelling, and 1 and 12 percent for stiffness. Abscesses were too infrequent to allow meaningful analysis of the relation to covariates, but for the other types of reactions we used logistic regression to examine the relation of occurrence to the delivery method (rifle or CO2-powered blowgun), adjuvant (FCA, Freund's modified adjuvant, and Freund's incomplete adjuvant), dart trauma (normal or abnormal), and age of mare. Abnormal dart trauma included cases where the dart hit bone or the needle broke off. We found strong evidence (odds ratio = 5.023, P = 0.001) for a higher probability of occurrence of swelling when darts were delivered by blowgun. We found some evidence (odds ratio = 8.729, P = 0.07) that abnormal dart trauma led to a higher frequency of nodule formation. Nodules were the most common reactions observed and often persisted for a year or more, but in our observations they did not appear to change any animal's range of movement or locomotor patterns and in most cases did not appear to differ in magnitude from naturally occurring injuries or scars. We were unable to perform histological examinations of these nodules, but they may be similar to granulomas reported by other investigators following administration of Freund's adjuvant.

Assessment LOPU-IVF in Japanese sika deer (Cervus nippon nippon) and application to Vietnamese sika deer (Cervus nippon pseudaxis) a related subspecies threatened with extinction.

PubMed

Locatelli, Y; Hendriks, A; Vallet, J-C; Baril, G; Duffard, N; Bon, N; Ortiz, K; Scala, C; Maurel, M-C; Mermillod, P; Legendre, X

2012-12-01

In mammals, recovery of oocytes by laparoscopic ovum pick-up (LOPU) coupled with in vitro production (IVP) of embryos represents a promising strategy for both amplification and genetic management of sparse animals from captive endangered wild species. As integrated technique developed mainly for domestic livestock, LOPU-IVP requires several studies to set up protocols for follicular stimulation or optimization of IVP before envisaging successful transposition to wild species. In deer, many endangered subspecies would be potentially concerned by applying such an approach using common subspecies for protocols optimization. The aim of the present study was to assess efficiency of follicle stimulation using ovine FSH (oFSH) for recovery of oocytes by LOPU in common sika deer (Cervus nippon nippon) before transposition of an optimized methodology for IVP of embryos from endangered Vietnamese sika deer hinds (Cervus nippon pseudaxis). In common sika deer, two doses of oFSH (0.25 and 0.5 U) and two frequencies of administration (12 and 24 h) were compared by monitoring of subsequent ovarian response, quality of oocytes recovered by LOPU, and in vitro developmental competence. In a first experiment, the dose of oFSH administered did not significantly affect the total number of follicles aspirated per hind per session (8.6 ± 1.0 vs. 8.2 ± 1.6 with 0.5 vs. 0.25 U oFSH, respectively; not significant). In a second experiment, frequency of 0.25 U oFSH administration did not affect ovarian response. Efficiency of IVP determined on blastocysts rates after in vitro maturation, fertilization, and development in oviduct epithelial cells coculture was increased when FSH was administered at 12-h intervals. Immune response after several follicular stimulations was detected against exogenous oFSH in plasma from the majority of sika deer hinds but was not associated with decreased ovarian response. When 0.25 U oFSH was administered at 12-h intervals to Vietnamese sika deer (N = 4), good quality cumulus oocyte complexes with complete and compact cumulus investments were recovered allowing a high cleavage rate after in vitro maturation and fertilization. Development to the blastocyst stage occurred in a high proportion (30% of oocytes) after coculture with ovine epithelial cells allowing cryobanking of transferable embryos from Vietnamese sika deer. These results confirm that LOPU-IVF after ovarian stimulation with oFSH may be a successful tool for cryobanking transferable embryos from endangered sika deer subspecies. Copyright © 2012 Elsevier Inc. All rights reserved.

Improving in vitro maturation and pregnancy outcome in cattle using a novel oocyte shipping and maturation system not requiring a CO₂ gas phase.

PubMed

Barceló-Fimbres, M; Campos-Chillón, L F; Mtango, N R; Altermatt, J; Bonilla, L; Koppang, R; Verstegen, J P

2015-07-01

The present work evaluated the benefit of a novel shipping and maturation medium (SMM) not requiring a CO2 gas for maturation and subsequent embryonic development of slaughterhouse and ovum pickup (OPU) bovine cumulus-oocyte complexes (COCs). Four experiments were conducted. In experiment 1, COCs were maturated for 18 hours in SMM and then incubated for 6 hours in, or 24 hours in a conventional system (control). Experiment 2 compared maturation for 24 hours in SMM versus 24 hours in the control. Experiment 3 compared three different incubation temperatures (37 °C, 38 °C, and 38.5 °C) for COCs maturation in SMM. In experiment 4, COCs obtained from 166 OPU sessions (representing two dairy and two beef breeds) in two locations (Wisconsin and California) were matured in SMM or control and evaluated relative to embryo production and pregnancy rates. Frozen semen was used for all experiments. The results for experiment 1 showed that the blastocyst rate and total embryo production rate (TE, Day-7 morulae plus all blastocysts) were higher for SMM than those in the control. However, no differences were observed for cleavage rate or blastocyst stage. In experiment 2, the blastocyst rate and TE were higher for SMM than those in the control; however, there was no difference for cleavage rate, total cell number, blastocyst stage. In experiment 3, the cleavage rate was similar, but the blastocyst rate and TE were greater for 38.5 °C than those for 38.0 °C and 37.5 °C. For experiment 4, Wisconsin OPU-derived COCs had a greater cleavage rate, blastocyst rate, TE, and blastocyst stage for SMM versus control. There were no breed effects. For the California trial, OPU-derived COCs matured in SMM had similar cleavage and pregnancy rates at Day 35 but greater blastocyst rates and transferred embryos per session than the control, which resulted in 2.2 more pregnancies per OPU session. Holstein COCs had superior embryonic development but similar pregnancy compared with Jersey. We conclude that COCs matured in SMM had greater oocyte competence than the control. Also, maturation at 38.5 °C in SMM was optimal for embryonic development. In summary, SMM resulted in greater embryonic development, similar pregnancy rates, but higher pregnancies per OPU session than the conventional maturation system. Copyright © 2015 Elsevier Inc. All rights reserved.

Effects of radiocobalt irradiation of unfertilized or fertilized rabbit OVA in vitro on subsequent fertilization and development in vivo

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chang, M C; Hunt, D M; Romanoff, E B

Freshly shed rabbit ova recovered from the Fallopian tubes were irradiated with a radiocobalt source at 45 r to 32,000 r and then transplanted into the left tube of mated rabbits. The ova were recovered and examined microscopically 10 hours to 24 hours, two days and 6 days after transplantation for the determination of fertilization, cleavage, and blastocyst formation. The fetus and uterine contents were examined macroscopically 22 to 25 days after transplantation. The proportion of fertilized ova decreased from 71% at a very low dosage, 45 r, to 46% at a very high dosage, 32,000 r. The proportion ofmore » normally cleaved ova and normal lilastocysts decreased from about 95% at 45 r, to about 3% at 500 r, to 0% at 6,500 r. The proportion of embryonic development decreased from 49% at 45 r, to 21% at 90 r, to 0% at 800 r. A chromosomal bridge was observed in an ovum irradiated at 6,500 r. Failure of second polar body division in one out of 23 ova irradiated at 6,500 r and polyspermy in one out of 32 ova irradiated at 32,000 r before fertilization was observed. When fertilized rabbit ova at the two cell stage were irradiated at 45 r to 6,500 r and examined at various times after transplantation, it was found that the proportion of normal cleavage decreased from 78% at 45 r to 33% at 800 r, to 0% at 6,500 r. The proportion of normal blastocysts decreased from 61% at 45 r, to 20% at 800 r, to 0% at 6,500 r. The proportion of normal embryonic development decreased from 46% at 45 r, to 12% at 500 r, to 0% at 6,500 r. In combination with data from a previous study of the irraiation of rabbit sper matozoa in vitro the following points are revealed: No abnormal fetus, no high proportion of degeneration after implantation, and no disturbance of the sex ratio were observed whether spermatozoa, or ova, unfertilized, or fertilized, were irradiated from 45 r to 800 r. Although there may be a differential sensitivity to various dosages for the subsequent cleavage and blastocyst formation following the irradiation of spermatozoa, unfertilized or fertilized ova at 45 r to 6,500 r, as far as subsequent embryonic development is concerned, the spermatozoa are more radioresistant than either unfertilized or fertilized ova and the unfertilized ova are more radiosensitive than fertilized ova. The chemical constituents of gametes necessary for the future development of the zygotes are more radiosensitive than are those for their fertilization and other activities.« less

Effects of Liuwei Dihuang Granule ([symbols; see text]) on the outcomes of in vitro fertilization pre-embryo transfer in infertility women with Kidney-yin deficiency syndrome and the proteome expressions in the follicular fluid.

PubMed

Lian, Fang; Wu, Hai-cui; Sun, Zhen-gao; Guo, Ying; Shi, Lei; Xue, Ming-yue

2014-07-01

To observe the effects of Liuwei Dihuang Granule ([symbols; see text], LDG) for tonifying Kidney (Shen) on the outcomes of in vitro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidney-yin deficiency syndrome and to explore its mechanism by detecting the proteome expression in the follicular fluid. Sixty-six infertility patients of Kidney-yin deficiency syndrome who would undergo IVF-ET, were randomly assigned to a treatment group and a control group according to a random number table, 33 cases in each group. Another 33 cases of non-Kidney-yin deficiency syndrome was taken as a syndrome-control group. Besides Western routine therapy, LDG was given 3 menstrual cycles before IVF to the treatment group, and a placebo granule to the control and syndrome-control groups. The scores of Kidney-yin deficiency symptoms (sore waist and knees, dry vagina, dysphoria with feverish sensation in the chest, palms and soles, etc.) were assessed, the number of retrieved oocytes, rates of high quality oocytes and embryos, fertility rate and clinical pregnancy rate were recorded, and the follicular fluid was collected on the day when the ovum was picked up, the differential protein expression was detected using two-dimensional gel electrophoresis, and then, matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins. The syndrome score in the treatment group decreased significantly from 16.09±2.58 to 8.67±2.13, while it changed insignificantly in the control group, with a significant difference in the lowering score between the two groups (P<0.05); the high quality rates of oocytes and embryos and clinical pregnancy rate were all superior in the treatment group to the control group (82.29% vs 78.08%, 76.76% vs 68.79%, 63.64% vs 36.36%, all P<0.05). The protein expression map from the follicular fluid showed that compared with the control group, 33 differential protein expressions were found in the syndrome-control group, among which 18 were down-regulated, and 15 up-regulated; in the treatment group 28 differential protein expressions were found, among which 15 were down-regulated, and 13 up-regulated. Through MALDI-TOF-MS, 14 proteins were identified (P<0.05). For the infertility patients undergoing IVF, LDG could alleviate clinical symptoms, improve rates of high quality oocytes and embryos, so as to raise clinical pregnancy rate. The mechanism may be through regulating proteome expression in the follicular fluid to improve the developmental microenvironment for oocytes which would lead to a successful embryo implantation.

Pupils teach to pupils about genetics or global warming

NASA Astrophysics Data System (ADS)

Cuny, Delphine

2013-04-01

The idea of this project is to put pupils in a teaching situation. Classes of teenagers go to primary schools and animate a science workshop. Junior pupils are separated in small groups and they attend two different sessions in the same half-day. The whole workshop consists of 4 sessions. Each session is organized with an activity (microscope observation, counting of chromosomes, drawing of a curve, etc.) in which senior pupils coach the younger, and ends with a debate or an assessment. The first experiment of this type of project was realized with a class of 14 to 15 year old pupils on the theme: How do your parents transmit your characteristics? The four sessions are attended in disorder but when knowledge of other sessions are necessary, senior pupils explain them at the beginning of the session. Junior pupils have a notebook to write their activities and to note their conclusions. Session 1: What did my father give to make me? Drawing and measuring microscopic observations of human spermatozoons. Conclusion: my father gave a spermatozoon which measures less than one mm long, this spermatozoon met my mother's egg and it made my first cell. Session 2: What does the program that made me look like? Microscope observation of blood cells, identification of chromosomes in the core. On microscope pictures, counting of chromosomes. Conclusion: My program is in each cell of my body, inside the core. Sometimes, in this core, we can observe short sticks that are called chromosomes. All human beings have the same number of chromosomes in their cells: 46. Session 3: Where do my chromosomes come from? Counting of chromosomes in spermatozoons or ovums and playing with sets of chromosomes to deduct sex of a baby. Conclusion: Daddy gave me 23 chromosomes and mummy gave me 23 chromosomes too. My program is then constituted from half of daddy's program and half of mummy's program. My brothers and sisters also have half and half, but not the same halves! Session 4: Where is the program that made me situated? Virtual experiments on the first cell of rats (core transfers) Conclusion: The program that made that a rat is itself is situated in the core of its first cell. It is called a genetic program. Second experiment of this type of project is realized with a class of 16-17 year old pupils, on global warming for 8 to 11 year old pupils from the neighbor school. The older pupils use a teaching set created by "la main à la pâte" foundation, the set is called "le climat, ma planète et moi" (the climate, my planet and me, http://www.fondation-lamap.org/fr/climat). This project is to take place in March 2013.

Effects of supplemental calcium salts of palm oil and chromium-propionate on insulin sensitivity and productive and reproductive traits of mid- to late-lactating Holstein × Gir dairy cows consuming excessive energy.

PubMed

Leiva, T; Cooke, R F; Brandão, A P; Bertin, R D; Colombo, E A; Miranda, V F B; Lourenço, L A C; Rodrigues, S M B; Vasconcelos, J L M

2018-01-01

This experiment compared insulin sensitivity, milk production, and reproductive outcomes in dairy cows consuming excessive energy during mid to late lactation and receiving in a 2 × 2 factorial design (1) concentrate based on ground corn (CRN; n = 20) or including 8% (DM basis) of Ca salts of palm oil (CSPO; n = 20), and (2) supplemented (n = 20) or not (n = 20) with 2.5 g/d of Cr-propionate. During the experiment (d 0-203), 40 multiparous, nonpregnant, lactating 3/4 Holstein × 1/4 Gir cows (initial days in milk = 81 ± 2; mean ± SE) were offered corn silage for ad libitum consumption, and individually received concentrate formulated to allow diets to provide 160% of their daily net energy for lactation requirements. From d -15 to 203, milk production was recorded daily, blood samples collected weekly, and cow body weight (BW) and body condition score (BCS) recorded on d 0 and 203. For dry matter intake evaluation, cows from both treatments were randomly divided in 5 groups of 8 cows each, and allocated to 8 individual feeding stations for 3 d. Intake was evaluated 6 times/group. Glucose tolerance tests (GTT; 0.5 g of glucose/kg of BW) were performed on d -3, 100, and 200. Follicle aspiration for in vitro embryo production was performed via transvaginal ovum pick-up on d -1, 98, and 198. Mean DMI, net energy for lactation intake, as well as BW and BCS change were similar across treatments. On average, cows gained 40 kg of BW and 0.49 BCS during the experiment. Within weekly blood samples, CRN cows had lower serum concentrations of glucose, insulin, fatty acids, and insulin-to-glucose ratio compared with CSPO cows, suggesting increased insulin sensitivity in CRN cows. During the GTT, insulin-sensitivity traits were also greater in CRN versus CSPO cows. Supplemental Cr-propionate resulted in lower serum insulin concentrations and insulin-to-glucose ratio within CRN cows only, indicating that Cr-propionate improved basal insulin sensitivity in CRN but not in CSPO cows. During the GTT, however, Cr-propionate supplementation reduced hyperinsulinemia and insulin-to-glucose ratio across CSPO and CRN cows. Milk production, as well as number of viable oocytes collected and embryos produced within each aspiration, were not affected by treatments. Hence, replacing corn by Ca salts of palm oil in the concentrate did not improve insulin sensitivity in Holstein × Gir dairy cows consuming excessive energy during mid to late lactation, whereas Cr-supplementation was effective in improving basal insulin sensitivity in cows not receiving Ca salts of palm oil. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Introducing reproductive life plan-based information in contraceptive counselling: an RCT.

PubMed

Stern, J; Larsson, M; Kristiansson, P; Tydén, T

2013-09-01

Can reproductive life plan (RLP)-based information in contraceptive counselling before pregnancy increase women's knowledge of reproduction, and of the importance of folic acid intake in particular? The RLP-based information increased women's knowledge of reproduction including knowledge of folic acid intake. Many women have insufficient knowledge of reproduction, including a health-promoting lifestyle prior to conception, and highly educated women in particular postpone childbearing until an age when their fertile capacity has started to decrease. The study was an randomized controlled trial with one intervention group (IG) and two control groups (CG1, CG2). A sample size calculation indicated that 82 women per group would be adequate. Recruitment took place during 3 months in 2012 and 299 women were included. The women were randomized in blocks of three. All groups received standard care (contraceptive counselling, Chlamydia testing, cervical screening). In addition, women in the IG were given oral and written RLP-based information about reproduction. A total of 299 out of 338 (88%) Swedish-speaking women visiting a Student Health Centre were included (mean age 23 years); response rate was 88%. Before the counselling, women in the IG and the CG1 completed a baseline questionnaire, including questions about lifestyle changes in connection to pregnancy planning, family planning intentions and knowledge of reproduction (e.g. the fecundity of an ovum). At follow-up 2 months after inclusion, a structured telephone interview was performed in all groups (n = 262, 88% participation rate). There was no difference between the groups regarding the mean knowledge score at baseline. The IG scored higher at follow-up than at baseline (P < 0.001); the mean increased from 6.4 to 9.0 out of a maximum 20 points. The women in the CG1 scored no differently at follow-up than at baseline. The difference in the knowledge score between the IG and the two CGs was significant (P < 0.001), whereas no difference was shown between the two CGs. There was no difference between the groups at baseline regarding how many women could mention folic acid intake among the things to do when planning to get pregnant. At follow-up, 22% in the IG, 3% in CG1 and 1% in CG2 mentioned folic acid intake (P < 0.001). At follow-up, more women in the IG also wished to have their last child earlier in life (P < 0.001) than at baseline, while there was no difference in the CG1. As the study sample consisted of university students, it is possible that the effect of the intervention was connected to a high level of education and conclusions for all women of reproductive age should be drawn with caution. The provision of RLP-based information seems to be a feasible tool for promoting reproductive health. Study funding was received from the Faculty of Medicine, Uppsala University, Sweden. There are no conflicts of interest. ClinicalTrial.gov Identifier NCT01739101.

Cysteamine supplementation during in vitro maturation of slaughterhouse- and opu-derived bovine oocytes improves embryonic development without affecting cryotolerance, pregnancy rate, and calf characteristics.

PubMed

Merton, J S; Knijn, H M; Flapper, H; Dotinga, F; Roelen, B A J; Vos, P L A M; Mullaart, E

2013-09-01

Optimization of ovum pick up (OPU) followed by in vitro embryo production (IVP) is strongly driven by the needs of both beef and dairy cattle breeders to enhance genetic improvement. The rapidly growing use of genomic selection in cattle has increased the interest in using OPU-IVP technology to increase the number of embryos and offspring per donor, thus allowing enhanced selection intensity for the next generation. The aim of this study was to optimize embryo production through supplementation of cysteamine during in vitro maturation (IVM) and in vitro culture (IVC) of both slaughterhouse- and OPU-derived oocytes. The effects on embryo production and on embryo cryotolerance, post-transfer embryo survival, and calf characteristics, including gestation length, birth weight, perinatal mortality, and sex ratio were studied. In study 1, immature slaughterhouse-derived cumulus-oocyte complexes (COCs) were matured in IVM medium supplemented with or without 0.1 mM cysteamine, fertilized and cultured for 7 days in 0.5 ml SOFaaBSA. In study 2, cysteamine was present during both IVM (0.1 mM) and IVC (0.01, 0.05, 0.1 mM) from Days 1 to 4. In study 3, OPU-derived COCs were matured in medium supplemented with or without 0.1 mM cysteamine in a 2 × 2 factorial design (OPU week and cysteamine treatment). Embryos were evaluated for stage and grade on Day 7 and, depending on the number of transferable embryos and recipients available, the embryos were transferred either fresh or frozen-thawed at a later date. The presence of cysteamine during IVM significantly increased the embryo production rate with slaughterhouse-derived COCs (24.0% vs. 19.4%). The higher number of embryos at Day 7 was due to an increased number of blastocysts, whereas the distribution of embryos among different quality grades and cryotolerance was not affected. Embryo production rate was negatively affected when cysteamine was present during both the processes of IVM and IVC during Days 1 to 4 of culture (13.2%-19.3% vs. 26.4%). The presence of cysteamine during IVM of OPU-derived COCs also significantly increased the embryo production rate (34.4% vs. 23.4%). The higher number of embryos was again totally due to an increased number of blastocysts, whereas cryotolerance was not affected. The relative increase in embryo production rate was higher with OPU-derived oocytes compared with slaughterhouse-derived COCs (47% vs. 24%). This improvement resulted in a mean of 1.73 transferable embryos per OPU session compared with 1.06 in the absence of cysteamine. The presence of cysteamine did not affect pregnancy rate, gestation length, birth weight, perinatal mortality, and sex of calves born from either fresh or frozen-thawed embryos. This study reported that cysteamine supplementation during IVM greatly improved the efficiency and affectivity of an OPU-IVP program. Copyright © 2013 Elsevier Inc. All rights reserved.

Age of G-1 PLUS v5 embryo culture medium is inversely associated with birthweight of the newborn.

PubMed

Kleijkers, Sander H M; van Montfoort, Aafke P A; Smits, Luc J M; Coonen, Edith; Derhaag, Josien G; Evers, Johannes L H; Dumoulin, John C M

2015-06-01

Does age of G-1 PLUS v5 embryo culture medium affect IVF outcome? Birthweight of singletons born after IVF showed an inverse association with age of the embryo culture medium, while no association was found between age of culture medium and fertilization rate, embryonic development or ongoing pregnancy. It has been reported that IVF culture media can deteriorate during storage, which suggests that the capacity of culture media to support optimal embryo development decreases over time. Some animal studies showed an effect of storage time on embryo development, in contrast to other studies, while the effect of aging culture medium on IVF outcome in humans is unknown. We used data on outcome of 1832 IVF/ICSI cycles with fresh embryo transfer, performed in the period 2008-2012 to evaluate the association of fertilization rate, embryonic development, ongoing pregnancy and birthweight of singletons with age of the culture medium (Vitrolife AB G-1 PLUS v5). Age of the culture medium was calculated by subtracting the production date from the date of ovum retrieval. Data analysis included linear regression and logistic regression on continuous and categorical outcomes, respectively. Age of the culture medium was not associated with fertilization rate (P = 0.543), early cleavage rate (P = 0.155), percentage of embryos containing four or more cells on Day 2 (P = 0.401), percentage of embryos containing eight or more cells on Day 3 (P = 0.175), percentage of embryos with multinucleated blastomeres (P = 0.527), or ongoing pregnancy (P = 0.729). However, birthweight of the newborn was inversely associated with age of the medium (β = -3.6 g, SE: 1.5 g, P = 0.021), after controlling for possible confounders (day of embryo transfer, number of transferred embryos, child's gender, gestational age at birth, parity, pregnancy complications, maternal smoking, height and weight, and paternal height and weight) and the association was not biased by year of treatment, time since first opening of the bottle or batch variations. This indicates a difference of 234 g in birthweight of newborns for media with an age difference of 65 days. The results from this study may be specific for the G-1 PLUS v5 culture medium and extrapolation of the results to other media should be done with caution because of the differences in composition and shelf life. Age of G-1 PLUS v5 medium used to culture human embryos affects birthweight of the respective newborn. This could imply that the preimplantation embryo adapts to its in vitro environment with lasting in vivo consequences. Therefore, it is important that companies are transparent about the exact composition of their embryo culture media, which will allow IVF clinics to further investigate the effects of the media or media components on the health of IVF children. No funding and no competing interests declared. Not applicable. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Economic Evaluation of Three Frequently Used Gonadotrophins in Assisted Reproduction Techniques in the Management of Infertility in the Netherlands.

PubMed

Fragoulakis, Vassilis; Pescott, Chris P; Smeenk, Jesper M J; van Santbrink, Evert J P; Oosterhuis, G Jur E; Broekmans, Frank J M; Maniadakis, Nikos

2016-12-01

Subfertility represents a multidimensional problem associated with significant distress and impaired social well-being. In the Netherlands, an estimated 50,000 couples visit their general practitioner and 30,000 couples seek medical specialist care for subfertility. We conducted an economic evaluation comparing recombinant human follicle-stimulating hormone (follitropin alfa, r-hFSH, Gonal-F ® ) with two classes of urinary gonadotrophins-highly purified human menopausal gonadotrophin (hp-HMG, Menopur ® ) and urinary follicle-stimulating hormone (uFSH, Fostimon ® )-for ovarian stimulation in women undergoing in vitro fertilization (IVF) treatment in the Netherlands. A pharmacoeconomic model was developed, simulating each step in the IVF protocol from the start of therapy until either a live birth, a new IVF treatment cycle or cessation of IVF, following a long down-regulation protocol. A decision tree combined with a Markov model details progress through each health state, including ovum pickup, fresh embryo transfer, up to two subsequent cryo-preserved embryo transfers, and (ongoing) pregnancy or miscarriage. A health insurer perspective was chosen, and the time horizon was set at a maximum of three consecutive treatment cycles, in accordance with Dutch reimbursement policy. Transition probabilities and costing data were derived from a real-world observational outcomes database (from Germany) and official tariff lists (from the Netherlands). Adverse events were considered equal among the comparators and were therefore excluded from the economic analysis. A Monte Carlo simulation of 5000 iterations was undertaken for each strategy to explore uncertainty and to construct uncertainty intervals (UIs). All cost data were valued in 2013 Euros. The model's structure, parameters and assumptions were assessed and confirmed by an external clinician with experience in health economics modelling, to inform on the appropriateness of the outcomes and the applicability of the model in the chosen setting. The mean total treatment costs were estimated as €5664 for follitropin alfa (95 % UI €5167-6151), €5990 for hp-HMG (95 % UI €5498-6488) and €5760 for uFSH (95 % UI €5256-6246). The probability of a live birth was estimated at 36.1 % (95 % UI 27.4-44.3 %), 33.9 % (95 % UI 26.2-41.5 %) and 34.1 % (95 % UI 25.9-41.8 %) for follitropin alfa, hp-HMG and uFSH, respectively. The costs per live birth estimates were €15,674 for follitropin alfa, €17,636 for hp-HMG and €16,878 for uFSH. Probabilistic sensitivity analysis indicated a probability of 72.5 % that follitropin alfa is cost effective at a willingness to pay of €20,000 per live birth. The probabilistic results remained constant under several analyses. The present analysis shows that follitropin alfa may represent a cost-effective option in comparison with uFSH and hp-HMG for IVF treatment in the Netherlands healthcare system.

Transgenerational healing: Educating children in genesis of healthy children, with focus on nutrition, emotion, and epigenetic effects on brain development.

PubMed

House, Simon H

2013-01-01

Although our continuing evolution can never achieve our perfection, we long for our children's birth and health to be near-perfect. Many children are born healthy, though fewer than is possible. Birthing and health rapidly improved generally due to modern housing, sanitation and medicine, as well as birth interventions. Arguably interventions have exceeded the optimal level, without enough regard for natural physical and intuitive resources. Conception, often too easy, receives too little personal preparation unless a couple has problems. Nurturing the health of sperm and ovum seems hard to focus on, yet is needed by both parents - and even by the four grandparents. What are the key factors? Positive: The fields of hormones/emotions and of nutrition/metabolism. Negative: stress, poor nutrition, toxins, diseases; much being due to poverty. Positive and negative both have structural and also epigenetic effects. Interventions, essential or inessential, are seldom without negative side effects. Health can best, and most economically, be generated at the beginning of life, through healthy conception, gestation and birth. Understanding prime needs improves initial health. It also informs therapy of any early-life problems. Healing is therefore more efficient when transgenerational, and much more powerful than individual healing. My vision of healing is safeguarding our evolution in progress. Children's choices - eating, exercise, emotional attitudes and relationships - are already profoundly affecting any children they may have, their mental and physical health. The most practical starting point seems to be educating boys as well as girls. Childhood is therefore the time to educate them in choices. The correction of often unnoticed problems- nutrient deficits, toxins, uro-genital disease - has enabled nearly nine out of ten couples to bear fully healthy babies, even following severe problems - infertility, miscarriages, stillbirths and malformations. Correcting problems before conception prevents both structural faults and wrong setting of gene-switches. Children's habits set. Once courting most are preoccupied and many pregnant unintentionally. Childhood is the time to be adopting a healthy lifestyle, the way to healthy babies The mother's nutritional and emotional status throughout pregnancy continues to affect her child's future physical and mental health, behaviour and ability. Before conception a woman needs to build her appropriate body stores - vitamins and minerals, proteins, docosahexaenoic acid. Before bearing another child, a replenishment time of 3 years is desirable. A return to childbearing in the 20s and early 30s could reduce risks that have risen with the recent shift towards conception by school children and by women in their late 30s or more. Governments, schoolteachers, health professionals, need to adopt this policy of transgenerational health. Empowerment with knowledge is the one way to fend off the growing pandemic of mental ill health and related disorders and to make the most of a nation's genetic potential. Financially there could be no better investment, let alone in enhancing people's lives. Childhood is the most appropriate time for education in this way to generating a healthy, able and peaceful human race. Essential to our amazing genetic systems are the resources of land, sea and air. We are one with our biosphere. We need urgently to follow up the vital work of Developmental Origins of Health and Disease, and of Far East initiatives in sea-bed and sea husbandry. © The Author(s) 2013.

Early pregnancy angiogenic markers and spontaneous abortion: an Odense Child Cohort study.

PubMed

Andersen, Louise B; Dechend, Ralf; Karumanchi, S Ananth; Nielsen, Jan; Joergensen, Jan S; Jensen, Tina K; Christesen, Henrik T

2016-11-01

Spontaneous abortion is the most commonly observed adverse pregnancy outcome. The angiogenic factors soluble Fms-like kinase 1 and placental growth factor are critical for normal pregnancy and may be associated to spontaneous abortion. We investigated the association between maternal serum concentrations of soluble Fms-like kinase 1 and placental growth factor, and subsequent spontaneous abortion. In the prospective observational Odense Child Cohort, 1676 pregnant women donated serum in early pregnancy, gestational week <22 (median 83 days of gestation, interquartile range 71-103). Concentrations of soluble Fms-like kinase 1 and placental growth factor were determined with novel automated assays. Spontaneous abortion was defined as complete or incomplete spontaneous abortion, missed abortion, or blighted ovum <22+0 gestational weeks, and the prevalence was 3.52% (59 cases). The time-dependent effect of maternal serum concentrations of soluble Fms-like kinase 1 and placental growth factor on subsequent late first-trimester or second-trimester spontaneous abortion (n = 59) was evaluated using a Cox proportional hazards regression model, adjusting for body mass index, parity, season of blood sampling, and age. Furthermore, receiver operating characteristics were employed to identify predictive values and optimal cut-off values. In the adjusted Cox regression analysis, increasing continuous concentrations of both soluble Fms-like kinase 1 and placental growth factor were significantly associated with a decreased hazard ratio for spontaneous abortion: soluble Fms-like kinase 1, 0.996 (95% confidence interval, 0.995-0.997), and placental growth factor, 0.89 (95% confidence interval, 0.86-0.93). When analyzed by receiver operating characteristic cut-offs, women with soluble Fms-like kinase 1 <742 pg/mL had an odds ratio for spontaneous abortion of 12.1 (95% confidence interval, 6.64-22.2), positive predictive value of 11.70%, negative predictive value of 98.90%, positive likelihood ratio of 3.64 (3.07-4.32), and negative likelihood ratio of 0.30 (0.19-0.48). For placental growth factor <19.7 pg/mL, odds ratio was 13.2 (7.09-24.4), positive predictive value was 11.80%, negative predictive value was 99.0%, positive likelihood ratio was 3.68 (3.12-4.34), and negative likelihood ratio was 0.28 (0.17-0.45). In the sensitivity analysis of 54 spontaneous abortions matched 1:4 to controls on gestational age at blood sampling, the highest area under the curve was seen for soluble Fms-like kinase 1 in prediction of first-trimester spontaneous abortion, 0.898 (0.834-0.962), and at the optimum cut-off of 725 pg/mL, negative predictive value was 51.4%, positive predictive value was 94.6%, positive likelihood ratio was 4.04 (2.57-6.35), and negative likelihood ratio was 0.22 (0.09-0.54). A strong, novel prospective association was identified between lower concentrations of soluble Fms-like kinase 1 and placental growth factor measured in early pregnancy and spontaneous abortion. A soluble Fms-like kinase 1 cut-off <742 pg/mL in maternal serum was optimal to stratify women at high vs low risk of spontaneous abortion. The cause and effect of angiogenic factor alterations in spontaneous abortions remain to be elucidated. Copyright © 2016 Elsevier Inc. All rights reserved.

[Elucidation of the mechanism of fertilization and clinical application of assisted reproductive technology].

PubMed

Hiroi, M

1996-08-01

Fertilization is the process including many events such as maturation of egg and sperm, attachment, binding, acrosomal reaction, penetration, fusion, cortical reaction, zona reaction and nuclear fusion of both gamete, whereby individual gametes from the female and male unite to create offspring. Although the reason for mechanism of fertilization is still not clearly understood, this process may accelerate the rate adaptation in evolution. In this special lecture, I would like to present our experimental and clinical results especially concerning with morphological, physiological, biochemical and molecular approach on the mechanism of fertilization. 1. Development and maturation of follicles and oocytes. It is well known that pituitary FSH, LH control the ovarian function. Follicular development and ovum maturation are also controlled by both pituitary gonadotropins and local factors such as autocrine and paracrine agents. When hMG is injected during 1-6 day of menstrual cycle, several dominant follicles are developed. If hMG is injected after selection of dominant follicles, only one dominant follicle develop in the ovary. When PMS-treated immature rats were injected with immature or mature follicle fluids, rats injected with mature follicular fluid showed strongly suppress in the ovarian weights and numbers of ovulated follicles. Also mature follicle suppress aromatization from and androstenedione to estradiol. These findings mean that mature follicular fluid contains inhibitory factors. Apoptosis of granulosa cells and follicular steroids are related to fertilization. 2. Intracellular calcium of oocyte. Intracellular calcium concentration is known to start to increase in a periodic manner after fertilization in oocytes of mammalians. In 65% of tested mouse oocytes, fertilization occurred during 4 hours observation after sperm insemination in vitro. An initial long lasting intracellular calcium concentration was observed and followed by periodic manner. This calcium oscillation is inhibited by calcium blockers such as verpamil and nifedipine, but increased by high concentration of extracellular calcium concentration in the medium. Role of increase of intracellular calcium are understood to prevent polysperm and activate metabolism of oocytes. 3. Glucose metabolism of oocytes. Mouse embryo utilizes pyruvate as an essential nutrient until the 8-cell stage, and glucose thereafter. We have devised non-radiometrie and enzymatic microassay method to measure glucose, deoxyglucose, deoxyglucose 6-phosphate incorporated into individual mouse oocytes and preimplantation embryo. In parallel, the activities of several enzymes of glycolytic pathway were also determined. In this study, glucose metabolism is necessary to develop in fertilized ova with changing activity of enzymes. 4. Molecular bases of ovarian fluid. The zona pellucida ZP is involved in a number of events in fertilization, all these fertilization events occur in the oviduct. Oviductal glycoprotein 200-240 KD has been identified from oviductal zona pellucida. Monoclonal antibody of oviductal glycoprotein reacted with ZP of oviductal egg but not with the ovarian egg. Anti-ZPO antibody inhibit to bind sperm to ZP. Sequences in mouse and hamster oviduct specific glycoprotein are estimated, this glycoprotein mRNA was observed in only oviduct by northern blotting method. These molecular gene expression was observed by in situ hybridization in the oviduct of estrous cycle of hamster. 5. Microinsemination of sperm. Microinsemination of sperm into oocyte is widely used in clinical medicine. Sperm penetration assay (hamster test) is useful method to estimate fertilization capacity of sperm. But immotile sperm cannot estimate it. So modified micro sperm penetration assay was established to estimate fertilization capacity of sperm by using micro-manipulator. Subzonal sperm injection (SUZI) and intracytoplasmic sperm injection (ICSI) promotes fertilization and cleavage rate in immotile

Interval of gonadotropin administration for in vitro embryo production from oocytes collected from Holstein calves between 2 and 6 months of age by repeated laparoscopy.

PubMed

Baldassarre, Hernan; Currin, Luke; Michalovic, Laura; Bellefleur, Anne-Marie; Gutierrez, Karina; Mondadori, Rafael G; Glanzner, Werner G; Schuermann, Yasmin; Bohrer, Rodrigo C; Dicks, Naomi; Lopez, Rosalba; Grand, François-Xavier; Vigneault, Christian; Blondin, Patrick; Gourdon, Jim; Bordignon, Vilceu

2018-08-01

Laparoscopic Ovum Pick-Up (LOPU) in calves followed by in vitro embryo production (IVEP) and transfer (ET) into adult recipients has great potential for accelerated genetic gain through shortening of the generation interval. In this study, 11 Holstein calves were subjected to up to six LOPU procedures between the ages of 2-6 months at 2-3 weeks interval. In all cases, the animals received a CIDR 5 days prior to LOPU and were gonadotropin-stimulated starting at 72 h before LOPU using one of three protocols that were rotated twice among the animals during the study. Calves were injected with FSH every 12 h (FSH12h), or every 8 h (FSH8h) or every 8 h until -36 h from LOPU at which point the FSH was replaced with a single dose of 400 IU eCG (FSH8h-eCG). No statistical differences were observed among the 3 treatments in terms of mean follicles available for aspiration (35.7 ± 16 vs. 38.5 ± 25 vs. 31.1 ± 22), mean oocytes recovered (26.5 ± 14 vs. 21.6 ± 10 vs. 19.4 ± 14) and cleavage rate (66.0 ± 14 vs. 61.1 ± 11 vs. 72.2 ± 8), for FSH12h, FSH8h and FSH8h-eCG, respectively. However, FSH8h-eCG resulted in a significantly higher rate of transferable embryos (17.5 ± 8%) compared with FSH12h (8.9 ± 5%, P < 0.05). Oocytes from follicles of ≥5 mm in diameter yielded a higher rate (P < 0.05) of development to the blastocyst stage (13.8%) than those collected from <5 mm follicles (6.8%). Animal age, by comparing animals at <100, 101 to 130 and > 130 days of age, did not affect the mean number of follicles (34.2 ± 15 vs. 39.3 ± 26 vs. 31.6 ± 25), the mean number of oocytes recovered (21.2 ± 10 vs. 24.5 ± 15 vs. 22.6 ± 17), and the cleavage rate (68.6 ± 11 vs. 61.7 ± 12 vs. 70.7 ± 10%), respectively. However, animals in the older age range had significantly higher development to the blastocyst stage (19.9 ± 6 vs. 9.5 ± 8%, P < 0.01) and better embryo quality, as evidenced by higher average cell numbers (119.1 ± 47 vs. 91.5 ± 25, P < 0.05) compared with those in the lower age. Finally, we tested the benefits of relieving endoplasmic reticulum stress by supplementing the culture medium with 50 μM tauroursodeoxycholic acid (TUDCA) and found a numerically higher rate of development to the blastocyst stage (21.1 ± 8 vs. 18.6 ± 4%), but not statistically different, compared with control culture. Overall, our findings indicate that a significant number of transferable embryos (range 10-30) can be produced from Holstein calves before they reach 6 months of age. Copyright © 2018 Elsevier Inc. All rights reserved.

Interventions for the prevention of OHSS in ART cycles: an overview of Cochrane reviews.

PubMed

Mourad, Selma; Brown, Julie; Farquhar, Cindy

2017-01-23

Ovarian hyperstimulation syndrome (OHSS) in assisted reproductive technology (ART) cycles is a treatment-induced disease that has an estimated prevalence of 20% to 33% in its mild form and 3% to 8% in its moderate or severe form. These numbers might even be higher for high-risk women such as those with polycystic ovaries or a high oocyte yield from ovum pickup. The objective of this overview is to identify and summarise all evidence from Cochrane systematic reviews on interventions for prevention or treatment of moderate, severe and overall OHSS in couples with subfertility who are undergoing ART cycles. Published Cochrane systematic reviews reporting on moderate, severe or overall OHSS as an outcome in ART cycles were eligible for inclusion in this overview. We also identified Cochrane submitted protocols and title registrations for future inclusion in the overview. The evidence is current to 12 December 2016. We identified reviews, protocols and titles by searching the Cochrane Gynaecology and Fertility Group Database of Systematic Reviews and Archie (the Cochrane information management system) in July 2016 on the effectiveness of interventions for outcomes of moderate, severe and overall OHSS. We undertook in duplicate selection of systematic reviews, data extraction and quality assessment. We used the AMSTAR (Assessing the Methodological Quality of Systematic Reviews) tool to assess the quality of included reviews, and we used GRADE methods to assess the quality of the evidence for each outcome. We summarised the characteristics of included reviews in the text and in additional tables. We included a total of 27 reviews in this overview. The reviews were generally of high quality according to AMSTAR ratings, and included studies provided evidence that ranged from very low to high in quality. Ten reviews had not been updated in the past three years. Seven reviews described interventions that provided a beneficial effect in reducing OHSS rates, and we categorised one additional review as 'promising'. Of the effective interventions, all except one had no detrimental effect on pregnancy outcomes. Evidence of at least moderate quality indicates that clinicians should consider the following interventions in ART cycles to reduce OHSS rates.• Metformin treatment before and during an ART cycle for women with PCOS (moderate-quality evidence).• Gonadotrophin-releasing hormone (GnRH) antagonist protocol in ART cycles (moderate-quality evidence).• GnRH agonist (GnRHa) trigger in donor oocyte or 'freeze-all' programmes (moderate-quality evidence). Evidence of low or very low quality suggests that clinicians should consider the following interventions in ART cycles to reduce OHSS rates.• Clomiphene citrate for controlled ovarian stimulation in ART cycles (low-quality evidence).• Cabergoline around the time of human chorionic gonadotrophin (hCG) administration or oocyte pickup in ART cycles (low-quality evidence).• Intravenous fluids (plasma expanders) around the time of hCG administration or oocyte pickup in ART cycles (very low-quality evidence).• Progesterone for luteal phase support in ART cycles (low-quality evidence).• Coasting (withholding gonadotrophins) - a promising intervention that needs to be researched further for reduction of OHSS.On the basis of this overview, we must conclude that evidence is currently insufficient to support the widespread practice of embryo cryopreservation. Currently, 27 reviews in the Cochrane Library were conducted to report on or to try to report on OHSS in ART cycles. We identified four review protocols but no new registered titles that can potentially be included in this overview in the future. This overview provides the most up-to-date evidence on prevention of OHSS in ART cycles from all currently published Cochrane reviews on ART. Clinicians can use the evidence summarised in this overview to choose the best treatment regimen for individual patients - a regimen that not only reduces the chance of developing OHSS but does not compromise other outcomes such as pregnancy or live birth rate. Review results, however, are limited by the lack of recent primary studies or updated reviews. Furthermore, this overview can be used by policymakers in developing local and regional protocols or guidelines and can reveal knowledge gaps for future research.

Reduced live-birth rates after IVF/ICSI in women with previous unilateral oophorectomy: results of a multicentre cohort study.

PubMed

Lind, Tekla; Holte, Jan; Olofsson, Jan I; Hadziosmanovic, Nermin; Gudmundsson, Johannes; Nedstrand, Elizabeth; Lood, Mikael; Berglund, Lars; Rodriguez-Wallberg, Kenny

2018-02-01

Is there a reduced live-birth rate (LBR) after IVF/ICSI treatment in women with a previous unilateral oophorectomy (UO)? A significantly reduced LBR after IVF/ICSI was found in women with previous UO when compared with women with intact ovaries in this large multicentre cohort, both crudely and after adjustment for age, BMI, fertility centre and calendar period and regardless of whether the analysis was based on transfer of embryos in the fresh cycle only or on cumulative results including transfers using frozen-thawed embryos. Similar pregnancy rates after IVF/ICSI have been previously reported in case-control studies and small cohort studies of women with previous UO versus women without ovarian surgery. In all previous studies multiple embryos were transferred. No study has previously evaluated LBR in a large cohort of women with a history of UO. This research was a multicentre cohort study, including five reproductive medicine centres in Sweden: Carl von Linné Clinic (A), Karolinska University Hospital (B), Uppsala University Hospital (C), Linköping University Hospital (D) and Örebro University Hospital (E). The women underwent IVF/ICSI between January 1999 and November 2015. Single embryo transfer (SET) was performed in approximately 70% of all treatments, without any significant difference between UO exposed women versus controls (68% versus 71%), respectively (P = 0.32), and a maximum of two embryos were transferred in the remaining cases. The dataset included all consecutive treatments and fresh and frozen-thawed cycles. The exposed cohort included 154 women with UO who underwent 301 IVF/ICSI cycles and the unexposed control cohort consisted of 22 693 women who underwent 41 545 IVF/ICSI cycles. Overall, at the five centres (A-E), the exposed cohort underwent 151, 34, 35, 41 and 40 treatments, respectively, and they were compared with controls of the same centre (18 484, 8371, 5575, 4670 and 4445, respectively). The primary outcome was LBR, which was analysed per started cycle, per ovum pick-up (OPU) and per embryo transfer (ET). Secondary outcomes included the numbers of oocytes retrieved and supernumerary embryos obtained, the Ovarian Sensitivity Index (OSI), embryo quality scores and cumulative pregnancy rates. We used a Generalized Estimating Equation (GEE) model for statistical analysis in order to account for repeated treatments. The exposed (UO) and control women's groups were comparable with regard to age and performance of IVF or ICSI. Significant differences in LBR, both crude and age-adjusted, were observed between the UO and control groups: LBR per started cycle (18.6% versus 25.4%, P = 0.007 and P = 0.014, respectively), LBR/OPU (20.3% versus 27.1%, P = 0.012 and P = 0.015, respectively) and LBR/ET (23.0% versus 29.7%, P = 0.022 and P = 0.025, respectively). The differences in LBR remained significant after inclusion of both fresh and frozen-thawed transfers (both crude and age-adjusted data): LBR/OPU (26.1% versus 34.4%, P = 0.005 and P = 0.006, respectively) and LBR/ET (28.3% versus 37.1%, P = 0.006 and P = 0.006, respectively). The crude cancellation rate was significantly higher among women with a history of UO than in controls (18.9% versus 14.5%, P = 0.034 and age-adjusted, P = 0.178). In a multivariate GEE model, the cumulative odds ratios for LBR (fresh and frozen-thawed)/OPU (OR 0.70, 95% CI 0.52-0.94, P = 0.016) and LBR (fresh and frozen-thawed)/ET (OR 0.68, 95% CI 0.51-0.92, P = 0.012) were approximately 30% lower in the group of women with UO when adjusted for age, BMI, reproductive centre, calendar period and number of embryos transferred when appropriate. The OSI was significantly lower in women with a history of UO than in controls (3.6 versus 6.0) and the difference was significant for both crude and age-adjusted data (P = <0.001 for both). Significantly fewer oocytes were retrieved in treatments of women with UO than in controls (7.2 versus 9.9, P = <0.001, respectively). Due to the nature of the topic, this is a retrospective analysis, with all its inherent limitations. Furthermore, the cause for UO was not possible to obtain in all cases. A diagnosis of endometriosis was also more common in the UO group, i.e. a selection bias in terms of poorer patient characteristics in the UO group cannot be completely ruled out. However, adjustment for all known confounders did not affect the general results. To date, this is the largest cohort investigated and the first study indicating an association of achieving reduced live birth after IVF/ICSI in women with previous UO. These findings are novel and contradict the earlier notion that IVF/ICSI treatment is not affected, or is only marginally affected by previous UO. None. Not applicable. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com