scholarly journals The Metabolic Signature of In Vitro Produced Bovine Embryos Helps Predict Pregnancy and Birth after Embryo Transfer

Metabolites ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 484
Author(s):  
Isabel Gimeno ◽  
Pablo García‐Manrique ◽  
Susana Carrocera ◽  
Cristina López‐Hidalgo ◽  
Luis Valledor ◽  
...  
Keyword(s):  
Amino Acids ◽  
Area Under The Curve ◽  
Culture Conditions ◽  
Embryonic Stage ◽  
Data Sets ◽  
Aconitic Acid ◽  
Characteristic Area ◽  
Oviduct Fluid ◽  
Pregnancy And Birth

In vitro produced (IVP) embryos show large metabolic variability induced by breed, culture conditions, embryonic stage and sex and gamete donors. We hypothesized that the birth potential could be accurately predicted by UHPLC-MS/MS in culture medium (CM) with the discrimination of factors inducing metabolic variation. Day-6 embryos were developed in single CM (modified synthetic oviduct fluid) for 24 h and transferred to recipients as fresh (28 ETs) or frozen/thawed (58 ETs) Day-7 blastocysts. Variability was induced with seven bulls, slaughterhouse oocyte donors, culture conditions (serum + Bovine Serum Albumin [BSA] or BSA alone) prior to single culture embryonic stage records (Day-6: morula, early blastocyst, blastocyst; Day-7: expanding blastocyst; fully expanded blastocysts) and cryopreservation. Retained metabolite signals (6111) were analyzed as a function of pregnancy at Day-40, Day-62 and birth in a combinatorial block study with all fixed factors. We identified 34 accumulated metabolites through 511 blocks, 198 for birth, 166 for Day-62 and 147 for Day-40. The relative abundance of metabolites was higher within blocks from non-pregnant (460) than from pregnant (51) embryos. Taxonomy classified lipids (12 fatty acids and derivatives; 224 blocks), amino acids (12) and derivatives (3) (186 blocks), benzenoids (4; 58 blocks), tri-carboxylic acids (2; 41 blocks) and 5-Hydroxy-l-tryptophan (2 blocks). Some metabolites were effective as single biomarkers in 95 blocks (Receiver Operating Characteristic – Area Under the Curve [ROC-AUC]: 0.700–1.000). In contrast, more accurate predictions within the largest data sets were obtained with combinations of 2, 3 and 4 single metabolites in 206 blocks (ROC-AUC = 0.800–1.000). Pregnancy-prone embryos consumed more amino acids and citric acid, and depleted less lipids and cis-aconitic acid. Big metabolic differences between embryos support efficient pregnancy and birth prediction when analyzed in discriminant conditions.

scholarly journals Model Distribution Effects on Likelihood Ratios in Fire Debris Analysis

Separations ◽  
2018 ◽  
Vol 5 (3) ◽  
pp. 44 ◽  
Author(s):  
Alyssa Allen ◽  
Mary Williams ◽  
Nicholas Thurn ◽  
Michael Sigman
Keyword(s):  
Computational Models ◽  
Area Under The Curve ◽  
Ground Truth ◽  
Data Sets ◽  
Likelihood Ratios ◽  
Data Set ◽  
Discriminant Model ◽  
Fire Debris ◽  
Characteristic Area ◽  
Ignitable Liquid

Computational models for determining the strength of fire debris evidence based on likelihood ratios (LR) were developed and validated against data sets derived from different distributions of ASTM E1618-14 designated ignitable liquid class and substrate pyrolysis contributions using in-silico generated data. The models all perform well in cross validation against the distributions used to generate the model. However, a model generated based on data that does not contain representatives from all of the ASTM E1618-14 classes does not perform well in validation with data sets that contain representatives from the missing classes. A quadratic discriminant model based on a balanced data set (ignitable liquid versus substrate pyrolysis), with a uniform distribution of the ASTM E1618-14 classes, performed well (receiver operating characteristic area under the curve of 0.836) when tested against laboratory-developed casework-relevant samples of known ground truth.

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

1998 ◽  
Vol 10 (3) ◽  
pp. 279 ◽  
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.

Review: Role of tubal environment in preimplantation embryogenesis: application to co-culture assays

Zygote ◽  
2010 ◽  
Vol 19 (1) ◽  
pp. 47-54 ◽  
Author(s):  
Pierre Guérin ◽  
Yves Ménézo
Keyword(s):  
Amino Acids ◽  
Embryo Culture ◽  
Metabolic Flux ◽  
Cultured Cells ◽  
Culture Media ◽  
Culture Conditions ◽  
Antioxidant Compounds ◽  
Beneficial Effects ◽  
Stage Embryo

SummaryThe culture of early preimplantation stage embryo is still delicate and the metabolic pathways of embryos are not completely understood. Embryo needs are evolutionary during the preimplantation development, consequently it is difficult to meet embryo needs in vitro. Culture conditions have to respect several physical and chemical equilibria: such as redox potential, pH, osmotic pressure, metabolic flux of energetic compounds, endogenous pools of amino acids and transcripts, etc. Embryo culture media are generally supplemented with amino acids, glucose, other energetic metabolites and antioxidant compounds, vitamin, and growth factors etc. Furthermore autocrine and paracrine regulation of embryo development probably exist. In fact embryo culture conditions have to be as non-toxic as possible. Various types of co-culture systems have been devised to overcome these problems. Complex interrelations exist between embryos and co-cultured cells. The beneficial effects of co-cultured cells may be due to continuous modifications of the culture medium, i.e. the elimination of toxic compounds and/or the supply of embryotrophic factors.

149 Bovine embryo selection can be improved by the characteristics of secreted extracellular vesicles

2019 ◽  
Vol 31 (1) ◽  
pp. 199
Author(s):  
E. Mellisho ◽  
M. Briones ◽  
F. O. Castro ◽  
L. Rodriguez-Alvarez
Keyword(s):  
Receiver Operating Characteristic ◽  
Predictive Power ◽  
Operating Characteristic ◽  
Culture Media ◽  
Area Under The Curve ◽  
Blastocyst Stage ◽  
Blastocyst Quality ◽  
Characteristic Area ◽  
Receiver Operating

Extracellular vesicles (EV) secreted by blastocysts might be relevant to predict competence of embryos produced in vitro. The aim of this study was to develop a model to select competent embryos that combines blastocyst morphokinetics data and morphological parameters of EV secreted during blastulation (Days 5-7.5). Embryos were cultured in groups up to Day 5; morulae were selected and individually cultured in SOFaa depleted of EV until Day 7.5 after IVF. Embryo competence was determined by in vitro post-hatching development up to Day 11. A retrospective classification of blastocyst and culture media was performed based on blastulation time [early (EB) or late (LB)] and competence at Day 11 [competent (C) or non-competent (NC)]. The EV were isolated from culture media of individual embryos, their properties determined by nanoparticle tracking analysis. The model was based on a binary logistic regression to describe the dichotomous-dependent variable of the blastocyst (C=1 and NC=0). A set of independent variables of blastocyst morphokinetics (blastulation time, blastocyst stage, blastocyst quality and blastocyst diameter at Day 7.5) and EV morphological parameters [mean size (ME), mode size (MO) and particle concentration (CO)] were analysed with multiple regression. The analysis generated the coefficients and their standard errors and significance level of an equation to calculate a probability, where values between 0.5 and 1 predict competent embryos. To verify the predictive power of the algorithm, the following indicators were used: the receiver operating characteristic with the determination of area under the curve, percentage correct predictions, and Omnibus tests. Statistical significance was determined at the P&lt;0.05 level. A rough guide for classifying the accuracy of a predictive model is as follows: 0.9 to 1=excellent, 0.8 to 0.9=good, 0.7 to 0.8=fair, 0.6 to 0.7=poor, 0.5 to 0.6=fail. A total of 254 embryos were used in this study; from them, 73 were classified in C-EB, 68 in NC-EB, 61 in C-LB and 52 in NC-LB. Initially, all independent variables were analysed in model 1; the most significant predictors associated with embryo competence were blastocyst stage, blastocyst quality, blastocyst diameter, ME and CO (P&lt;0.05). In model 2 no significant variables were excluded (blastulation time and MO). The statistical test of predictive power indicates that models 1 and 2 achieved a receiver operating characteristic-area under the curve of 0.853 (95% confidence interval, 0.806-0.9; P&lt;0.001) and correct predictions of 77.2 and 77.6%, respectively. When EV characteristics were excluded and the model considers only variables from the embryo, the receiver operating characteristic-area under the curve value was 0.714 (95% confidence interval, 0.651-0.777; P&lt;0.001) and correct predictions was reduced to 65.4. Model 2 was consider the most appropriate from the practical point of view because it avoids disturbing embryo culture during blastulation. The results indicate that incorporating EV properties increases accuracy of embryo selection, supporting the possibility to improve conventional methods by combining blastocyst morphology and characteristics of EV obtained by nanoparticle tracking analysis. This work was supported by Fondecyt 1170310.

scholarly journals Accounting for biases in riboprofiling data indicates a major role for proline and not positive amino acids in stalling translation

2014 ◽  
Author(s):  
Carlo G. Artieri ◽  
Hunter B. Fraser
Keyword(s):  
Amino Acids ◽  
Ribosome Profiling ◽  
Side Chain ◽  
Data Sets ◽  
Charged Amino Acids ◽  
Recent Method ◽  
Low Coverage ◽  
Positively Charged

The recent advent of ribosome profiling ? sequencing of short ribosome-bound fragments of mRNA ? has offered an unprecedented opportunity to interrogate the sequence features responsible for modulating translational rates. Nevertheless, numerous analyses of the first riboprofiling dataset have produced equivocal and often incompatible results. Here we analyze three independent yeast riboprofiling data sets, including two with much higher coverage than previously available, and find that all three show substantial technical sequence biases that confound interpretations of ribosomal occupancy. After accounting for these biases, we find no effect of previously implicated factors on ribosomal pausing. Rather, we find that incorporation of proline, whose unique side-chain stalls peptide synthesis in vitro, also slows the ribosome in vivo. We also reanalyze a recent method that reported positively charged amino acids as the major determinant of ribosomal stalling and demonstrate that its assumptions lead to false signals of stalling in low-coverage data. Our results suggest that any analysis of riboprofiling data should account for sequencing biases and sparse coverage. To this end, we establish a robust methodology that enables analysis of ribosome profiling data without prior assumptions regarding which positions spanned by the ribosome cause stalling.

Metabolism of pyruvate by pre-elongation sheep embryos and effect of pyruvate and lactate concentrations during culture in vitro

1993 ◽  
Vol 5 (4) ◽  
pp. 417 ◽  
Author(s):  
JG Thompson ◽  
AC Bell ◽  
PA Pugh ◽  
HR Tervit
Keyword(s):  
Amino Acids ◽  
Significant Interaction ◽  
Essential Amino Acids ◽  
Blastocyst Stage ◽  
Energy Substrates ◽  
Lactate Levels ◽  
Oviduct Fluid ◽  
High Lactate

In the first of two experiments, utilization of [1-14C]pyruvate by 8-cell and blastocyst-stage embryos derived in vivo was examined during a 3-h incubation in HEPES-buffered synthetic oviduct fluid (SOF) medium in the presence or absence of other substrates. In the second, a factorial design examined the effect of pyruvate (0, 0.33, 1.0 and 3.3 mM) and lactate (3.3, 10 and 33 mM) on development of 1- and 2-cell sheep embryos cultured in vitro in a modified SOF medium (containing glucose, glutamine and modified Eagle's medium non-essential amino acids). Peak utilization of [1-14C]pyruvate was unaffected by the presence or absence of other energy substrates. In contrast, rate of utilization was affected by the addition of other energy substrates, with half maximal utilization occurring at either 0.4 +/- 0.2 mM or 1.2 +/- 0.2 mM for 8-cells and either 0.2 +/- 0.2 mM or 1.3 +/- 0.3 mM for blastocysts when incubated in the absence or presence of other energy substrates respectively. In the second experiment the proportion of embryos developing to blastocysts was inhibited by high lactate levels (P < 0.001), but was generally not affected by pyruvate concentration. However, there was a significant interaction (P < 0.001) between pyruvate and lactate when both were present in the medium. At 0.33 mM pyruvate, 3.3 mM lactate supported good development (83 +/- 8% blastocysts) whereas 10 mM lactate supported less development (50 +/- 11%). However, at the higher levels of pyruvate this effect was lost.(ABSTRACT TRUNCATED AT 250 WORDS)

Neutrophil migration through in vitro interstitial matrix

1992 ◽  
Vol 50 (1) ◽  
pp. 894-895
Author(s):  
J. Roemer ◽  
S.R. Simon
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Cell Migration ◽  
Smooth Muscle Cells ◽  
Inflammatory Cell ◽  
Neutrophil Migration ◽  
Culture Conditions ◽  
Aortic Smooth Muscle ◽  
Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

Antimyotoxic Activity of Synthetic Peptides Derived from Bothrops atrox Snake Gamma Phospholipase A2 Inhibitor Selected by Virtual Screening

2019 ◽  
Vol 19 (22) ◽  
pp. 1952-1961 ◽  
Author(s):  
J.C. Sobrinho ◽  
A.F. Francisco ◽  
R. Simões-Silva ◽  
A.M. Kayano ◽  
J.J. Alfonso Ruiz Diaz ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Docking ◽  
Synthetic Peptides ◽  
Cell Damage ◽  
Neutralization Assay ◽  
Phospholipase Activity ◽  
Phospholipases A2 ◽  
Catalytically Active ◽  
Bothrops Atrox

Background: Several studies have aimed to identify molecules that inhibit the toxic actions of snake venom phospholipases A2 (PLA2s). Studies carried out with PLA2 inhibitors (PLIs) have been shown to be efficient in this assignment. Objective: This work aimed to analyze the interaction of peptides derived from Bothrops atrox PLIγ (atPLIγ) with a PLA2 and to evaluate the ability of these peptides to reduce phospholipase and myotoxic activities. Methods: Peptides were subjected to molecular docking with a homologous Lys49 PLA2 from B. atrox venom modeled by homology. Phospholipase activity neutralization assay was performed with BthTX-II and different ratios of the peptides. A catalytically active and an inactive PLA2 were purified from the B. atrox venom and used together in the in vitro myotoxic activity neutralization experiments with the peptides. Results: The peptides interacted with amino acids near the PLA2 hydrophobic channel and the loop that would be bound to calcium in Asp49 PLA2. They were able to reduce phospholipase activity and peptides DFCHNV and ATHEE reached the highest reduction levels, being these two peptides the best that also interacted in the in silico experiments. The peptides reduced the myotubes cell damage with a highlight for the DFCHNV peptide, which reduced by about 65%. It has been suggested that myotoxic activity reduction is related to the sites occupied in the PLA2 structure, which could corroborate the results observed in molecular docking. Conclusion: This study should contribute to the investigation of the potential of PLIs to inhibit the toxic effects of PLA2s.

Sign in / Sign up

Export Citation Format

Share Document