Preimplantation genetic screening (PGS) is widely used to select in vitro-fertilized embryos free of chromosomal abnormalities and to improve the clinical outcome of in vitro fertilization (IVF). A ...disadvantage of PGS is that it requires biopsy of the preimplantation human embryo, which can limit the clinical applicability of PGS due to the invasiveness and complexity of the process. Here, we present and validate a noninvasive chromosome screening (NICS) method based on sequencing the genomic DNA secreted into the culture medium from the human blastocyst. By using multiple annealing and looping-based amplification cycles (MALBAC) forwhole-genome amplification (WGA), we performed next-generation sequencing (NGS) on the spent culture medium used to culture human blastocysts (n = 42) and obtained the ploidy information of all 24 chromosomes. We validated these results by comparing each with their corresponding whole donated embryo and obtained a high correlation for identification of chromosomal abnormalities (sensitivity, 0.882, and specificity, 0.840). With this validated NICS method, we performed chromosome screening on IVF embryos from seven couples with balanced translocation, azoospermia, or recurrent pregnancy loss. Six of them achieved successful clinical pregnancies, and five have already achieved healthy live births thus far. The NICS method avoids the need for embryo biopsy and therefore substantially increases the safety of its use. The method has the potential of much wider chromosome screening applicability in clinical IVF, due to its high accuracy and noninvasiveness.
The aim of the present study was to validate methods of stool sample conservation for the egg hatch test (EHT). This study involved the use of a bovine naturally infected predominantly by Cooperia ...spp. and one equine naturally infected predominantly by cyathostomins characterized as susceptible to benzimidazoles in the EHT. Fecal samples were submitted to three treatments: aerobic methods (anaerobic storage in plastic bottles, anaerobic storage in vacuum-sealed bags or aerobic storage in plastic bags), under two temperature conditions (room temperature and refrigeration) analyzed at four different assessment times (48, 72, 96 and 120 h). As the standard test, an assay was also performed within 3 h. The tests were performed in triplicate for each drug concentration and with three experimental repetitions at one-week intervals. Two criteria were used for the storage methods: hatchability in the negative control group and sensitivity of the eggs to thiabendazole, comparing the EC50 and 95% confidence interval for each treatment to those of the standard test and the other repetitions. Bovine samples can be stored for up to 96 h and refrigerated vacuum storage can be used, ensuring hatchability of the negative control and sensitivity of the eggs to thiabendazole. For equine samples, no forms of storage were indicated due to the variation among the repetitions and the reduction in the sensitivity of the eggs to thiabendazole, which could result in a false positive detection of resistance.
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•Bovine feces can be stored for up to 96 h, but the EHT must be performed within 3 h for equine feces samples.•Samples stored at room temperature have good hatchability but with reduced sensitivity to thiabendazole in the EHT.•Vacuum-sealed refrigerated samples ensure adequate hatchability.
The utilization of lutein as a natural colorant or nutraceutical in many foods, supplements, and other commercial products is currently limited because of its low water-solubility and chemical ...instability. The purpose of this study was to evaluate the effect of Maillard conjugates on the physical and chemical stability of lutein-enriched emulsions exposed to different temperatures and pH values, as well as on their potential gastrointestinal fate. Oil-in-water emulsions were prepared using either casein or casein-dextran conjugates as emulsifiers. Both types of emulsions showed a slight increase in particle aggregation at temperatures exceeding 37 °C, and became more prone to color fading (lutein degradation) as the temperature was increased. Casein-coated oil droplets were highly unstable to flocculation near their isoelectric point (pH 4–5) due to the reduction in electrostatic repulsion. However, casein-dextran-coated droplets were stable from pH 3 to 7, which was attributed to strong steric repulsion by the dextran moiety. The casein-coated droplets were unstable to aggregation in the gastric phase of the simulated GIT, whereas the casein-dextrin-coated ones were stable, which was again attributed to increased steric repulsion. Emulsifier type did not strongly influence lutein bioaccessibility. This work shows that Maillard conjugates can improve the physical stability of lutein-enriched emulsions without adversely affecting the bioaccessibility of the bioactive agent.
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•Casein-dextran conjugates were formed using the Maillard reaction.•Lutein-enriched emulsions were stabilized by casein or casein-dextran conjugates.•Maillard conjugation inhibited isoelectric flocculation of casein-coated droplets.•Maillard conjugation did not inhibit color fading due to lutein degradation.•The use of Maillard conjugates did not affect lutein bioaccessibility.
Diet and fertility: a review Gaskins, Audrey J.; Chavarro, Jorge E.
American journal of obstetrics and gynecology,
04/2018, Letnik:
218, Številka:
4
Journal Article
Recenzirano
Odprti dostop
The literature on the relationship between diet and human fertility has greatly expanded over the last decade, resulting in the identification of a few clear patterns. Intake of supplemental folic ...acid, particularly at doses higher than those recommended for the prevention of neural tube defects, has been consistently related to lower frequency of infertility, lower risk of pregnancy loss, and greater success in infertility treatment. On the other hand and despite promising evidence from animal models, vitamin D does not appear to exert an important role in human fertility in the absence of deficiency. Antioxidant supplementation does not appear to offer any benefits to women undergoing infertility treatment, but it appears to be beneficial when it is the male partner who is supplemented. However, the available evidence does not allow discerning which specific antioxidants, or at which doses, are responsible for this benefit. Long-chain omega-3 fatty acids appear to improve female fertility, although it remains unclear to what extent contamination of shared food sources, such as fish with high levels of environmental toxicants, can dampen this benefit. Lastly, adherence to healthy diets favoring seafood, poultry, whole grains, fruits, and vegetables are related to better fertility in women and better semen quality in men. The cumulative evidence has also piled against popular hypotheses. Dairy and soy, once proposed as reproductive toxicants, have not been consistently related to poor fertility. In fact, soy and soy supplements appear to exert a beneficial effect among women undergoing infertility treatment. Similarly, because data from large, high-quality studies continue to accumulate, the evidence of a potentially deleterious effect of moderate alcohol and caffeine intake on the ability to become pregnant seems less solid than it once did. While a complete picture of the role of nutrition on fertility is far from complete, much progress has been made. The most salient gaps in the current evidence include jointly considering female and male diets and testing the most consistent findings in randomized trials.
In Vitro Embryo Production in Sheep Mondal, S; Mor, A; Reddy, I J ...
Methods in molecular biology (Clifton, N.J.),
2019, Letnik:
2006
Journal Article
The recent advances in biotechnological research have led to development of many advanced reproductive techniques and biological tools which are set to revolutionize the productive efficiency of ...livestock species. The development of technology for sequencing of whole genomes and mass screening of gene regulation has widened our approach to genetic profiling and mapping, as well as furthering our understanding of underlying physiological mechanisms. The newer biotechnologies of gene transfer, in vitro fertilization and embryo production, cloning, and stem cell technology have been developed and are being refined with efficiencies suitable for use in animal farming. Efficient in vitro systems for maturing oocytes, fertilizing, and developing embryos have resulted in commercial in vitro production of embryos. Here we describe in vitro maturation, in vitro fertilization, embryo production, embryo culture, and quantitation of gene expression in sheep embryos.
Aptamers are short oligonucleotides isolated in vitro from randomized libraries that can bind to specific molecules with high affinity, and offer a number of advantages relative to antibodies as ...biorecognition elements in biosensors. However, it remains difficult and labor‐intensive to develop aptamer‐based sensors for small‐molecule detection. Here, we review the challenges and advances in the isolation and characterization of small‐molecule‐binding DNA aptamers and their use in sensors. First, we discuss in vitro methodologies for the isolation of aptamers, and provide guidance on selecting the appropriate strategy for generating aptamers with optimal binding properties for a given application. We next examine techniques for characterizing aptamer–target binding and structure. Afterwards, we discuss various small‐molecule sensing platforms based on original or engineered aptamers, and their detection applications. Finally, we conclude with a general workflow to develop aptamer‐based small‐molecule sensors for real‐world applications.
Aptamers, oligonucleotide‐based recognition elements isolated from random libraries, have several favorable attributes for biosensing. This Review examines state‐of‐the‐art methods and advances in the isolation and characterization of small‐molecule‐binding aptamers and their use in various biosensors. Factors limiting aptamer‐based sensors and potential solutions to these issues are also discussed.
Fish oil is used for its numerous health and nutritional benefits. Nevertheless, it is unstable during production, storage, and application. Nanoemulsions can be used as an effective carrier for ...encapsulated nutraceuticals. The effects of different components on the stability of fish oil nanoemulsions with soybean protein isolate-phosphatidylcholine (SPI-PC) were studied by characterizing particle size, polydispersity index, ζ-potential, turbidity, and turbiscan stability index. The SPI-PC nanoemulsions with optimal stability were prepared with 2% SPI, 0.2% PC, 1.5% fish oil, and 100 MPa homogenization pressure. Confocal laser scanning microscopy verified that the oil droplet was encapsulated inside the nanoemulsions. Compared with a Tween 20 nanoemulsions control group, the SPI-PC nanoemulsions have better storage and oxidative stability, and have better resistance to certain concentrations (0.1–0.5 M) of Na+. However, we found that the SPI-PC nanoemulsions resistance to acidic conditions was not as good as that of Tween 20. The SPI-PC nanoemulsions showed aggregation of droplets during in vitro gastric digestion while the Tween 20 nanoemulsions did not. The release rate of free fatty acids (FFA) reached 86.8% in the 2 h in vitro intestine digestion. SPI-PC nanoemulsions significantly increased the digestibility of fish oil compared with Tween 20 nanoemulsions. Hence, SPI-PC nanoemulsions could be a good way to stabilize fish oil and improve digestibility under simulated gastrointestinal conditions.
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•The effect of different components on the stability of fish oil nanoemulsions stabilized with SPI-PC was studied.•Fish oil nanoemulsions stabilized with SPI-PC have good stability at different conditions.•The release rate of FFA of fish oil nanoemulsions stabilized with SPI-PC was 86.8%.
To study the possible transmission, to the next generation, of epigenetic defects associated with in vitro maturation (IVM) of human oocytes.
Case–control study using epigenetic data.
Two ...collaborating university departments.
Eleven IVM newborns and 19 controls, conceived by conventional assisted reproduction.
Chorionic villus and cord-blood sampling.
Using bisulfite pyrosequencing, we have measured average methylation levels of 6 imprinted (LIT1, MEG, MEST, NESPas, PEG3, and SNRPN), 5 tumor-suppressor (APC, ATM, BRCA1, RAD51C, and TP53), 2 pluripotency (NANOG and OCT4), and 2 metabolic (LEP and NR3C1) genes, as well as 2 repetitive elements (ALU and LINE1) in 2 tissues of IVM and control neonates. Using deep bisulfite sequencing, we have determined methylation patterns of many individual DNA molecules to detect rare RAD51C epimutations (allele methylation errors).
No statistically significant impact was found of IVM on chorionic villus and cord-blood DNA methylation at the studied developmentally important genes and interspersed repeats. The RAD51C epimutation rate was low (0.5% ± 0.1%) in all analyzed samples.
IVM-induced epigenetic changes in offspring, if any, are relatively small in magnitude and/or infrequent.
STUDY QUESTION
Is in vitro maturation (IVM) as successful as standard in vitro fertilization (IVF) for the treatment of patients with polycystic ovaries (PCO) in terms of fresh, frozen and cumulative ...pregnancy outcomes?
SUMMARY ANSWER
There was no difference in clinical pregnancy rates in fresh or frozen embryo transfer (FET) cycles between the two treatment groups however, the IVM group showed a lower clinical pregnancy rate cumulatively. There was significantly fewer live births resulting from IVM treatment for both fresh and cumulative cycle outcomes however, there was no difference in live birth rates resulting from FETs between IVM and IVF treatment.
WHAT IS KNOWN ALREADY
IVM is well recognized as the only treatment option to eliminate completely the incidence of ovarian hyperstimulation syndrome. However, historically IVM has been less successful than standard IVF in terms of clinical pregnancy, implantation and live birth rates.
STUDY DESIGN, SIZE, AND DURATION
This paper represents a retrospective case–control study. The study involved 121 participants who underwent 178 treatment cycles. Cycles were completed between March 2007 and December 2012. All fresh cycles and subsequent FET cycles were included in the analysis to calculate cumulative outcomes.
PARTICIPANTS/MATERIALS, SETTING, AND METHODS
All participants were prospectively diagnosed with PCO morphology or polycystic ovarian syndrome (PCOS) and underwent either IVM or standard IVF treatment. Their treatment outcomes were analysed with regard to embryological data, and the rate of biochemical pregnancy, clinical pregnancy and live birth, in addition maternal and neonatal outcomes were assessed. Fifty-six patients underwent 80 cycles of IVM treatment and 65 patients underwent 98 cycles of standard IVF treatment.
MAIN RESULTS AND THE ROLE OF CHANCE
For fresh cycles, the differences in the biochemical pregnancy, clinical pregnancy or miscarriage rates between the two treatment groups were not statistically significant. The IVM group showed significantly lower live birth rates in fresh cycles in comparison to standard IVF treatment (18.8 versus 31.0%, P = 0.021). For frozen embryo transfer (FET) cycles the differences in biochemical pregnancy, clinical pregnancy, live birth or miscarriage rates between the two treatments groups were not statistically significant. The cumulative biochemical pregnancy (67.5 versus 83.7%, P = 0.018), clinical pregnancy (51.3 versus 65.3%, P = 0.021) and live birth rates (41.3 versus 55.1%, P = 0.005) were significantly lower in the IVM group in comparison to the standard IVF treatment group. There was no overall difference in the cumulative miscarriage rates between the two treatment groups. There was no difference between treatment methods with regard to the neonatal outcomes, and the IVM group had a significantly lower rate of ovarian hyperstimulation syndrome (0 versus 7.1%, P < 0.001).
LIMITATIONS, REASONS FOR CAUTION
This was an observational study and further randomized clinical trials are required to clarify the difference in outcomes between standard IVF and IVM for patients with PCO/PCOS.
WIDER IMPLICATIONS OF THE FINDINGS
This is the first study to compare IVM with standard IVF in PCO/PCOS patients using blastocyst development and single embryo transfer. Furthermore, it is the first study to show the results of fresh, frozen and cumulative treatment cycle outcomes between the two groups. Our results show similar success rates to those reported from other groups, particularly in relation to the incidence of miscarriage in fresh IVM cycles and improved success from FET cycles. Maternal and neonatal outcomes are consistent with the limited literature available.
STUDY FUNDING/COMPETING INTEREST(S)
The study was supported by the Women's and Infant's Research Foundation of Western Australia. Professor Hart is Medical Director of Fertility Specialists of Western Australia (FSWA) and a shareholder Western IVF. He has received educational sponsorship from MSD, Merck-Serono and Ferring Pharmaceuticals. T.H. is a consultant with FSWA and a shareholder in Western IVF. She has received educational sponsorship from MSD, Merck-Serono and Ferring Pharmaceuticals. The other authors have no competing interests.
To determine if increasing paternal age has an adverse effect on pregnancy outcomes in paired donor egg recipients who received oocytes from the same donor in the same stimulation cycle.
...Retrospective cohort study.
Reproductive Medicine Center.
The study included 154 recipients who received oocytes from a split donor oocyte cycle and received sperm from men in discrepant age groups (group A: <45 years old; group B: ≥45 years old).
None.
Implantation rate, pregnancy loss rate, pregnancy rate, and live birth rate.
The median paternal age was 41 years old for group A and 48 years old for group B. The pregnancy rate was 81% in group A compared with 69% in group B. The live birth rate was 65% in group A compared with 53% in group B. The rate of pregnancy loss was 19% in group A and 23% in group B. The implantation rate was 69% in group A compared with 66% in group B. The adjusted odds of pregnancy were found to be 65% lower for patients in the older partner age group (95% confidence interval CI, 0.13, 0.95). The adjusted odds of live birth rate (odds ratio OR, 0.45; 95% CI, 0.20, 1.00), implantation rate (OR, 0.91; 95% CI, 0.43, 1.92), and rate of pregnancy loss (OR, 1.5; 95% CI, 0.5, 4.5) favored the younger partner age group; however, these results were not statistically significant.
In this model that controlled for oocyte quality to the greatest degree possible by using paired recipients from the same donor from the same stimulation cycle, we found that increased paternal age had a negative effect on pregnancy rates.
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