Comprehensive analysis of whole genome methylation in mouse blastocysts cultured with four different constituents following in vitro fertilization

Abstract Background With the development of assisted reproductive technology (ART), diseases believed to be caused by ART have begun to be identified as imprinted disease. However, no conclusion has been reached. So we sought to determine whether ART procedures disturb gene methylation and whether imprinted genes alone are selectively disturbed. To examine whether the constituents of the culture medium contribute to the changes in methylation, we used a mouse model to conduct IVF and comprehensively analyzed 5′–C–phosphate–G–3′ (CpG) by reduced representation bisulfite sequencing (RRBS) using a second-generation sequencer to determine changes in methylation using four types of culture media with different amino acid constituents. Results We cultured ova to the blastocyst stage in a mouse model in culture media with four different amino acid constituents. Each culture medium included (1) KSOM culture medium (NoAA), (2) KSOM media + essential amino acids (EAAs), (3) KSOM medium + non-essential amino acids (NEAAs), or (4) KSOM medium + EAAs + NEAAs (AllAA) analyzed by reduced representation bisulfite sequencing. The results showed that (1) there were many regions that maintained hypermethylation with NEAAs, (2) there was little effect of demethylation on reprogramming in the 5′UTR and promoter regions, and (3) specific changes were observed in imprinted genes such as Nnat and Nespas. Conclusions Compared with EAAs, NEAAs could protect genes from demethylation caused by reprogramming. On the imprinted genes, methylation of the promoter region of H19 was decreased by NEAAs, suggesting that specific genes were prone to changes in methylation. It was suggested that these changes could provide similar results in humans. Further studies are needed to understand how changes in methylation may affect gene expression profiles.

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P–283 Combination between amino acids profile of the spent culture media and morphokinetics parameters of human embryos to determine its viability

Human Reproduction ◽

10.1093/humrep/deab130.282 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

N Adel ◽

M Kadah ◽

S Abdulghafar ◽

M Elmahdy ◽

D Ghareeb ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Embryo Development ◽

Culture Medium ◽

Essential Amino Acids ◽

Human Embryos ◽

Data Set ◽

Group A ◽

Amino Acids Profile ◽

Group D

Abstract Study question How to determine human embryo viability noninvasively before embryo transfer? Summary answer We propose that the combination of the amino acid profile of an individual embryo with its morphokinetics will provide noninvasive tool to determine its viability. What is known already It was already known that human embryos at early cleavage require non-essential amino acids, while at the 8-cell to blastocyst stages, a mixture of non-essential and essential amino acids. Amino acids have important roles during embryo development. Acting as biosynthetic precursors,buffers of intracellular pH in the embryo, antioxidants, energy sources and regulators of metabolic function and signaling pathways. Many studies have used time-lapse to analyze human embryonic development including the process of fertilization and assessment of early events and introducednoninvasive prognostic markers which predict embryo development and correlate it to IVF treatment outcomes. Study design, size, duration This study was a prospective cohort study approved by the Clinical Trial Ethical Committee of Faculty of Medicine, Alexandria University according to ethical standards of scientific research (Serial number: 0303721).Thirty females aged 30.13 ± 4.83 years undergoing ICSI cycle in the Madina Fertility Center, during the period of March 2018 to November 2019.202 MII oocytes were incubated individually in embryoscope. Participants/materials, setting, methods Embryos (n = 161) were divided on Day 5 into two groups –developed embryos “Group D” (embryos that developed to blastocyst) and arrested embryos “Group A” (embryos remain at cleavage stage and fail to develop to blastocys).Developed embryos (Group D) included 99 embryos, and Arrested embryos (Group A) included 62 embryos. For each group, morphokinetic developmental points using embryoscope and the different amino acids concentrations in spent culture medium were analyzed using LC- mass spectro etry. Main results and the role of chance On one hand, the first appearance of pronuclei (TPNa), t2, t4 and CC2 in group D occurred significantly earlier than those of Group A.Analysis of 19 essential and non-essential amino acids in spent culture medium of each embryo in the two studied groups D and A showed a significantly higher concentration of two essential amino acids L-Valine (145.73 ± 150.96) and L-Phenylalanine (61.59 ± 55.78) in Group D than their concentration in Group A ( 104.58 ± 33.58, 44.24 ± 14.61, respectively , p ≤ 0.05).and significantly lower concentration of three non-essential amino acids L-Tyrosine (62.56 ± 41.03) , L-Cysteine (19.48 ± 11.90), and L-Alanine (136.0 ± 389.83) observed in Group D when compared to Group A (69.57 ± 20.78, 22.37 ± 8.59,145.33 ± 165.22, respectively, Limitations, reasons for caution It is important to note, that results were developed on a data set from one clinic with different stimulation protocols, a multicenter data and a correlation with the stimulation protocol used should be involved in future studies, in addition a larger sample size to avoid high standard deviation is recommended Wider implications of the findings: We can conclude that amino acid turnover is independent of the traditional morphological assessment of embryos and it may reflect its viability. The prospective combined use of amino acids profile of individual embryo and its morphokinetic parameters may contribute to introduce a new noninvasivs tool that may improve implantation rate Trial registration number 0303721

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Amino acid metabolism of bovine blastocysts derived from parthenogenetically activated or in vitro fertilized oocytes

Reproduction Fertility and Development ◽

10.1071/r98052 ◽

1998 ◽

Vol 10 (3) ◽

pp. 279 ◽

Cited By ~ 10

Author(s):

Y. G. Jung ◽

T. Sakata ◽

E. S. Lee ◽

Y. Fukui

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Metabolism ◽

Culture Medium ◽

Acid Metabolism ◽

Essential Amino Acids ◽

Fluid Medium ◽

Vitro Fertilization ◽

Oviduct Fluid

The uptake and synthesis of 19 amino acids by fresh or frozen–thawed bovine blastocysts produced by parthenogenesis (PT) or in vitro fertilization (IVF) were compared in the present study. Fresh blastocysts, 180 h after IVF or PT activation, and frozen–thawed blastocysts, 168 h old and cultured for 12 h post-thawing, were cultured in synthetic oviduct fluid medium (SOFM) containing polyvinyl alcohol (PVA) with both essential and non-essential amino acids (EAA and NEAA, respectively) (Medium 1: M1) or SOFM containing PVA with only EAA (Medium 2: M2). In Experiment 1, when fresh or frozen–thawed PT blastocysts were cultured in M1, the uptake of glutamate (in fresh only), aspartate and arginine, and the synthesis of glutamine and alanine were significantly enhanced. In the culture with M2, serine, asparagine, glutamate, glutamine, glycine, arginine and alanine were significantly taken up. It was found that the glutamine concentrations was significantly higher (P < 0.001) in the culture medium drops containing embryos than in the drops without embryos. In Experiment 2, when PT blastocysts were cultured in M1, the uptake of aspartate and synthesis of alanine were greater (P < 0.01) than those by IVF blastocysts. When M2 was used, a significant (P < 0.01) production of serine, asparagine, glutamate, glutamine and alanine, and the uptake of arginine by PT blastocysts were observed. In Experiment 3, when IVF blastocysts were cultured in M1, fresh blastocysts depleted more aspartate and glutamate, and produced more glutamine and alanine than frozen–thawed blastocysts. When cultured in M2, frozen–thawed blastocysts depleted more threonine (P < 0.01) than fresh blastocysts. These results indicate that the uptake and synthesis of amino acids were different in fresh or frozen–thawed bovine blastocysts derived from PT or IVF. These differences in amino acid metabolism may be related to the viability of the blastocysts.

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Effects of amino acids on development in vitro of cleavage-stage bovine embryos into blastocysts

Reproduction Fertility and Development ◽

10.1071/rd9960835 ◽

1996 ◽

Vol 8 (5) ◽

pp. 835 ◽

Cited By ~ 28

Author(s):

T Pinyopummintr ◽

BD Bavister

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Embryo Development ◽

Culture Medium ◽

Essential Amino Acids ◽

Bovine Embryo ◽

Bovine Embryos ◽

Development In Vitro ◽

Defined Culture

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.

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Amino Acid Metabolism in Human Embryos

Physiological Research ◽

10.33549/physiolres.933240 ◽

2016 ◽

pp. 823-832 ◽

Cited By ~ 3

Author(s):

P. DRÁBKOVÁ ◽

L. ANDRLOVÁ ◽

R. HAMPL ◽

R. KANĎÁR

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Metabolism ◽

Culture Medium ◽

Acid Metabolism ◽

Culture Media ◽

Time Lapse ◽

Human Embryos ◽

Strongly Correlated ◽

Cell Stage

The aim of this study was to find some relationship between amino acid metabolism and the embryo morphokinetic parameters studied via time-lapse analysis. Study included 48 human embryo samples and their culture media. Two groups of embryos were identified: embryos reached the 8-cell stage on day 3 (n=34) and embryos failed to develop at any point during the incubation (n=14). Amino acids levels were measured on day 3 of embryo development; using time-lapse analysis, the precise timing of embryo cleavage, synchrony of division, grade of fragmentation etc. were established. No statistically significant differences between dividing and arresting embryos were observed in terms of amino acids production/consumption and turnover. Amino acids which were part of the culture medium did not exhibit any statistically significant correlation with kinetic parameters with the exception of the grade of fragmentation on day 3; there were negative correlation with glutamate, and positive with glutamine, glycine and taurine. In some dividing and in some arresting embryos appeared new amino acids which strongly correlated with each other, with methionine, but not with any other amino acid that is a regular part of the culture medium.

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Very Low Ethanol Concentrations Affect the Viability and Growth Recovery in Post-Stationary-Phase Staphylococcus aureus Populations

Applied and Environmental Microbiology ◽

10.1128/aem.72.4.2627-2636.2006 ◽

2006 ◽

Vol 72 (4) ◽

pp. 2627-2636 ◽

Cited By ~ 20

Author(s):

Indranil Chatterjee ◽

Greg A. Somerville ◽

Christine Heilmann ◽

Hans-Georg Sahl ◽

Hans H. Maurer ◽

...

Keyword(s):

Amino Acids ◽

Staphylococcus Aureus ◽

Amino Acid ◽

Stationary Phase ◽

Culture Medium ◽

Culture Media ◽

Growth Media ◽

Cell Integrity ◽

Amino Acid Depletion

ABSTRACT Pharmaceuticals, culture media used for in vitro diagnostics and research, human body fluids, and environments can retain very low ethanol concentrations (VLEC) (≤0.1%, vol/vol). In contrast to the well-established effects of elevated ethanol concentrations on bacteria, little is known about the consequences of exposure to VLEC. We supplemented growth media for Staphylococcus aureus strain DSM20231 with VLEC (VLEC+ conditions) and determined ultramorphology, growth, and viability compared to those with unsupplemented media (VLEC− conditions) for prolonged culture times (up to 8 days). VLEC+-grown late-stationary-phase S. aureus displayed extensive alterations of cell integrity as shown by scanning electron microscopy. Surprisingly, while ethanol in the medium was completely metabolized during exponential phase, a profound delay of S. aureus post-stationary-phase recovery (>48 h) was observed. Concomitantly, under VLEC+ conditions, the concentration of acetate in the culture medium remained elevated while that of ammonia was reduced, contributing to an acidic culture medium and suggesting decreased amino acid catabolism. Interestingly, amino acid depletion was not uniformly affected: under VLEC+ conditions, glutamic acid, ornithine, and proline remained in the culture medium while the uptake of other amino acids was not affected. Supplementation with arginine, but not with other amino acids, was able to restore post-stationary-phase growth and viability. Taken together, these data demonstrate that VLEC have profound effects on the recovery of S. aureus even after ethanol depletion and delay the transition from primary to secondary metabolite catabolism. These data also suggest that the concentration of ethanol needed for bacteriostatic control of S. aureus is lower than that previously reported.

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Screening of amino acid constituents from date palm fruits

International Journal of Bioassays ◽

10.21746/ijbio.2016.10.0011 ◽

2016 ◽

Vol 5 (10) ◽

pp. 4972

Author(s):

Lata Birlangi

Keyword(s):

Amino Acids ◽

Amino Acid ◽

United Arab Emirates ◽

Nutritional Value ◽

Date Palm ◽

Amino Acid Content ◽

Essential Amino Acids ◽

Cultivated Plants ◽

Palm Fruits ◽

Social Part

The date palm (Phoenix dactylifera L.) is one of mankind’s oldest cultivated plants. The fruit of the date palm is an important crop of the hot arid and semi-arid regions of the world. It has always played a genuine economic and social part in the lives of the people of these areas. The present objective in examining the amino acid content of different varieties of date palm fruits from Middle-East region; is to determine whether its protein could effectively supplement the nutritional value and it is also aimed in finding which variety is rich in number of amino acids. The phytochemical screening revealed the presence of eight essential amino acids and five non-essential amino acids in the date fruits. Among all the date fruit varieties taken as samples for the study, Dabbas cultivar of United Arab Emirates found to exhibit eight types of amino acids which includes five as non-essential ones. Total of thirteen amino acids were detected in the seven date cultivars. Determination of amino acid can serve as a guide to the possible nutritional value.

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Progress towards reduced-crude protein diets for broiler chickens and sustainable chicken-meat production

Journal of Animal Science and Biotechnology ◽

10.1186/s40104-021-00550-w ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Sonia Yun Liu ◽

Shemil P. Macelline ◽

Peter V. Chrystal ◽

Peter H. Selle

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Broiler Chickens ◽

Crude Protein ◽

Plasma Concentrations ◽

Essential Amino Acids ◽

Chicken Meat ◽

Meat Production ◽

Accurate Identification ◽

Reduced Crude Protein

AbstractThe prime purpose of this review is to explore the pathways whereby progress towards reduced-crude protein (CP) diets and sustainable chicken-meat production may be best achieved. Reduced-CP broiler diets have the potential to attenuate environmental pollution from nitrogen and ammonia emissions; moreover, they have the capacity to diminish the global chicken-meat industry’s dependence on soybean meal to tangible extents. The variable impacts of reduced-CP broiler diets on apparent amino acid digestibility coefficients are addressed. The more accurate identification of amino acid requirements for broiler chickens offered reduced-CP diets is essential as this would diminish amino acid imbalances and the deamination of surplus amino acids. Deamination of amino acids increases the synthesis and excretion of uric acid for which there is a requirement for glycine, this emphasises the value of so-called “non-essential” amino acids. Starch digestive dynamics and their possible impact of glucose on pancreatic secretions of insulin are discussed, although the functions of insulin in avian species require clarification. Maize is probably a superior feed grain to wheat as the basis of reduced-CP diets; if so, the identification of the underlying reasons for this difference should be instructive. Moderating increases in starch concentrations and condensing dietary starch:protein ratios in reduced-CP diets may prove to be advantageous as expanding ratios appear to be aligned to inferior broiler performance. Threonine is specifically examined because elevated free threonine plasma concentrations in birds offered reduced-CP diets may be indicative of compromised performance. If progress in these directions can be realised, then the prospects of reduced-CP diets contributing to sustainable chicken-meat production are promising.

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Further evidence that mechanisms of host/symbiont integration are dissimilar in the maternal versus embryonic Acyrthosiphon pisum bacteriome

EvoDevo ◽

10.1186/s13227-020-00168-5 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Celeste R. Banfill ◽

Alex C. C. Wilson ◽

Hsiao-ling Lu

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Acyrthosiphon Pisum ◽

Essential Amino Acids ◽

Developmental Time ◽

Follicular Epithelium ◽

Future Research ◽

Buchnera Aphidicola ◽

Bacterial Endosymbionts ◽

Asexual Embryogenesis

Abstract Background Host/symbiont integration is a signature of evolutionarily ancient, obligate endosymbioses. However, little is known about the cellular and developmental mechanisms of host/symbiont integration at the molecular level. Many insects possess obligate bacterial endosymbionts that provide essential nutrients. To advance understanding of the developmental and metabolic integration of hosts and endosymbionts, we track the localization of a non-essential amino acid transporter, ApNEAAT1, across asexual embryogenesis in the aphid, Acyrthosiphon pisum. Previous work in adult bacteriomes revealed that ApNEAAT1 functions to exchange non-essential amino acids at the A. pisum/Buchnera aphidicola symbiotic interface. Driven by amino acid concentration gradients, ApNEAAT1 moves proline, serine, and alanine from A. pisum to Buchnera and cysteine from Buchnera to A. pisum. Here, we test the hypothesis that ApNEAAT1 is localized to the symbiotic interface during asexual embryogenesis. Results During A. pisum asexual embryogenesis, ApNEAAT1 does not localize to the symbiotic interface. We observed ApNEAAT1 localization to the maternal follicular epithelium, the germline, and, in late-stage embryos, to anterior neural structures and insect immune cells (hemocytes). We predict that ApNEAAT1 provisions non-essential amino acids to developing oocytes and embryos, as well as to the brain and related neural structures. Additionally, ApNEAAT1 may perform roles related to host immunity. Conclusions Our work provides further evidence that the embryonic and adult bacteriomes of asexual A. pisum are not equivalent. Future research is needed to elucidate the developmental time point at which the bacteriome reaches maturity.

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The screening and preliminary construction of quality mutant population for cultivar “Nipponbare” in japonica rice (Oryza sativa)

Biologia ◽

10.2478/s11756-012-0106-x ◽

2012 ◽

Vol 67 (6) ◽

Author(s):

Da Zhang ◽

Jian Wu ◽

Guan Li ◽

Chun Shi

Keyword(s):

Amino Acids ◽

Oryza Sativa ◽

Amino Acid ◽

Mutation Frequency ◽

Near Infrared ◽

Essential Amino Acids ◽

Japonica Rice ◽

Near Infrared Reflectance ◽

Mutant Population ◽

Nonessential Amino Acids

AbstractProgenies of Oryza sativa cv. Nipponbare induced with 0.4% ethyl methane sulphonate (EMS) were screened for quality mutants and the preliminary quality mutant population was constructed in present experiment. A total of 2210 materials were first screened using near infrared reflectance spectroscopy (NIRS) from which 208 quality mutants were obtained for a second screening and then yielded 73 quality mutants including amylase content (AC), gel consistency (GC), gelatinization temperature (GT), protein content (PC), rapid viscosity analysis (RVA) parameters and amino acid contents. The screening yielded 11 PC mutants with a mutation frequency of 4.98‰, followed by 7 rice floury viscosity mutants (3.17‰), 5 AC mutants (2.26‰), 4 chalky mutants, GT and GC mutants (1.81‰), and 2 ASV mutants (0.9‰). The relative contents of 17 kinds of amino acid mutations, including 7 kinds for essential amino acids and 10 kinds for nonessential amino acids were identified. With the variation of 10% as the screening standard, mutants were obtained for lysine and leucine at 0.45‰ and for valine at 4.98‰, but no mutants were found for isoleucine, phenylalanine, threonine. For nonessential amino acids, mutants of glutamic (0.45‰), arginine (3.62‰), alanine (3.17‰), serine (0.45‰), glycine (0.45‰), tyrosine (1.81‰), proline (2.71‰), and histidine (0.45‰) were obtained, but none was found for aspartic, phenylalanine nor threonine. At 100% as the screening standard for methionine and cysteines, the mutation frequency of these two amino acid mutants were 0.9‰ and 4.98‰ respectively. Quality mutants in this preliminary library of rice could play important role in gene function and breeding of rice quality.

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MATHEMATICAL MODELS FOR THE SYNTHESIS OF PLANT-BASED COMPOSITIONS WITH IMPROVED AMINO ACID COMPOSITION

10.32008/geolinks2021/b1/v3/12 ◽

2021 ◽

Author(s):

Irina Gaivoronskaya ◽

Valenitna Kolpakova

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Mathematical Models ◽

Functional Properties ◽

Amino Nitrogen ◽

Essential Amino Acids ◽

Major Allergen ◽

Structural Features ◽

Covalent Bonds ◽

Pea Protein

The aim of the work was to optimize the process of obtaining multicomponent protein compositions with high biological value and higher functional properties than the original vegetable protein products. Was realized studies to obtain biocomposites on the base of pea protein-oat protein and pea protein-rice protein. Developed composites were enriched with all limited amino acids. For each of the essential amino acids, the amino acid score was 100% and higher. Protein products used in these compositions are not in major allergen list, which allows to use these compositions in allergen-free products and specialized nutrition. To determine biosynthesis parameters for compositions from pea protein and various protein concentrates with the use of transglutaminase enzyme, was studied effect of concentration and exposition time on the amount of amino nitrogen released during the reaction. Decreasing of amino nitrogen in the medium indicated the occurrence of a protein synthesis reaction with the formation of new covalent bonds. Were determined optimal parameters of reaction: the hydromodule, the exposure time, the concentration of EP of the preparation, were obtained mathematical models. Studies on the functional properties of composites, the physicochemical properties of the proteins that make up their composition, and structural features will make it possible to determine the uses in the manufacture of food products based on their ability to bind fat, water, form foam, gels, and etc.

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