International legal framework for the criminal legal protection of the embryo

Legal Protection ◽

Legal Framework ◽

Criminal Code ◽

International Aspects

In connection with the active development and use of assisted reproductive technologies, protection of the human embryo and its legal status issue is currently being actualized. We make an attempt to reveal and explain some of the international aspects of the criminal law protection of the life and rights of the embryo. We consider the concept of “embryo” not only from the point of view of various scientific approaches (medicine, biology, embryology, jurisprudence), but also from the legislative side. We present and analyze the first mention of the embryo in Roman private law in connection with modern domestic law. We carry out an analysis of international legal acts that provide protection of embryos both “in vitro” and “in vivo”, followed by consideration of specific criminal law norms of foreign countries, namely Brazil and Colombia. We pay attention to some of the most famous cases from the jurisprudence of the European Court of Human Rights in order to understand the applied international legal acts “de facto”. The study also takes into account modern domestic legislation and considers point “g” of part 2 of Article 105 of the Criminal Code of the Russian Federation.

Head and neck tumors and assisted reproductive technologies: social and legal aspects

Head and neck tumors (HNT) ◽

10.17650/2222-1468-2020-10-3-90-96 ◽

2020 ◽

Vol 10 (3) ◽

pp. 90-96

Author(s):

N. A. Ognerubov

Keyword(s):

Head And Neck ◽

Legal Status ◽

Head And Neck Cancers ◽

Head And Neck Tumors ◽

Central Nervous System Tumors ◽

Neck Tumors

Background. Currently, the proportion of pregnancies obtained by assisted reproductive technologies (ARTs) reaches 1.7–4.0 %. Short-term and long-term results of ART implementation require public and, most importantly, legal assessment. Some publications suggest higher risks of congenital deformities and head and neck cancers in children conceived by ART.Objective: to review publications analyzing the problem of head and neck cancers in children conceived by ART and legal protection of the embryo.Materials and methods. We performed retrospective analysis of 42 articles published in 1995–2019, including 33 foreign and 9 Russian articles.Results. Earlier studies have demonstrated that children conceived by ART have an increased risk of cancers, including head and neck cancers, such as central nervous system tumors and retinoblastoma. Recent publications have shown no significant differences in the prevalence of malignant tumors between children born after ART and children conceived naturally. Nevertheless, the risk of developing head and neck tumors is higher in children after ART. The analysis of Russian and international legislation has demonstrated that the legal status of an embryo differs depending on whether it develops in vivo or in vitro.Conclusion. Children conceived by ART are at higher risk of malignant head and neck tumors, primarily central nervous system tumors and retinoblastoma. The legal status of an embryo depends on whether it develops in vivo or in vitro.

How to Achieve High-Quality Oocytes? The Key Role of Myo-Inositol and Melatonin

International Journal of Endocrinology ◽

10.1155/2016/4987436 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 36

Author(s):

Salvatore Giovanni Vitale ◽

Paola Rossetti ◽

Francesco Corrado ◽

Agnese Maria Chiara Rapisarda ◽

Sandro La Vignera ◽

...

Keyword(s):

Oocyte Quality ◽

Fertilization Rate ◽

In Vitro Models ◽

The One ◽

High Level

Assisted reproductive technologies (ART) have experienced growing interest from infertile patients seeking to become pregnant. The quality of oocytes plays a pivotal role in determining ART outcomes. Although many authors have studied how supplementation therapy may affect this important parameter for both in vivo and in vitro models, data are not yet robust enough to support firm conclusions. Regarding this last point, in this review our objective has been to evaluate the state of the art regarding supplementation with melatonin and myo-inositol in order to improve oocyte quality during ART. On the one hand, the antioxidant effect of melatonin is well known as being useful during ovulation and oocyte incubation, two occasions with a high level of oxidative stress. On the other hand, myo-inositol is important in cellular structure and in cellular signaling pathways. Our analysis suggests that the use of these two molecules may significantly improve the quality of oocytes and the quality of embryos: melatonin seems to raise the fertilization rate, and myo-inositol improves the pregnancy rate, although all published studies do not fully agree with these conclusions. However, previous studies have demonstrated that cotreatment improves these results compared with melatonin alone or myo-inositol alone. We recommend that further studies be performed in order to confirm these positive outcomes in routine ART treatment.

Reproductive fluids, used for the in vitro production of pig embryos, result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM.

10.21203/rs.3.rs-42227/v2 ◽

2020 ◽

Author(s):

Evelynne Paris-Oller ◽

Sergio Navarro-Serna ◽

Cristina Soriano-Úbeda ◽

Jordana Sena Lopes ◽

Carmen Matas ◽

...

Keyword(s):

Reproductive Performance ◽

Culture Media ◽

In Vitro Production ◽

Aberrant Expression ◽

Low Efficiency ◽

The Impact

Abstract Background: In vitro embryo production (IVP) and embryo transfer (ET) are two very common assisted reproductive technologies (ART) in human and cattle. However, in pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow. In addition, the potential impact of these two ART on the health of the offspring is unknown. We investigated here if the use of a modified IVP system, with natural reproductive fluids (RF) as supplements to the culture media, combined with a minimally invasive surgery to perform ET, affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits.Results: The blastocyst rates obtained by both in vitro systems, conventional (C-IVP) and modified (RF-IVP), were similar. Pregnancy and farrowing rates were also similar. However, when compared to in vivo control (artificial insemination, AI), litter sizes of both IVP groups were lower, while placental efficiency was higher in AI than in RF-IVP. Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI, but not for RF-IVP group.Conclusions: The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring.

DNA methylation and gene expression changes derived from assisted reproductive technologies can be decreased by reproductive fluids

eLife ◽

10.7554/elife.23670 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 49

Author(s):

Sebastian Canovas ◽

Elena Ivanova ◽

Raquel Romar ◽

Soledad García-Martínez ◽

Cristina Soriano-Úbeda ◽

...

Keyword(s):

Dna Methylation ◽

Cell Number ◽

Rna Seq ◽

Methylation Analysis ◽

Number Of Children ◽

Methylation Patterns

The number of children born since the origin of Assisted Reproductive Technologies (ART) exceeds 5 million. The majority seem healthy, but a higher frequency of defects has been reported among ART-conceived infants, suggesting an epigenetic cost. We report the first whole-genome DNA methylation datasets from single pig blastocysts showing differences between in vivo and in vitro produced embryos. Blastocysts were produced in vitro either without (C-IVF) or in the presence of natural reproductive fluids (Natur-IVF). Natur-IVF embryos were of higher quality than C-IVF in terms of cell number and hatching ability. RNA-Seq and DNA methylation analyses showed that Natur-IVF embryos have expression and methylation patterns closer to in vivo blastocysts. Genes involved in reprogramming, imprinting and development were affected by culture, with fewer aberrations in Natur-IVF embryos. Methylation analysis detected methylated changes in C-IVF, but not in Natur-IVF, at genes whose methylation could be critical, such as IGF2R and NNAT.

116 DEVELOPMENT OF ASSISTED REPRODUCTION TECHNOLOGIES FOR THE ENDANGERED MISSISSIPPI GOPHER FROG (RANA SEVOSA) AND SPERM TRANSFER FOR IN VITRO FERTILIZATION

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab116 ◽

2012 ◽

Vol 24 (1) ◽

pp. 170 ◽

Cited By ~ 11

Author(s):

A. Kouba ◽

E. Willis ◽

C. Vance ◽

S. Hasenstab ◽

S. Reichling ◽

...

Keyword(s):

Captive Breeding ◽

Sperm Concentration ◽

Critically Endangered ◽

Leopard Frog ◽

Genetic Management ◽

Mississippi Gopher Frog

Species-specific differences in breeding strategies and physiology have limited the application of assisted reproductive technologies (ART) for critically endangered amphibians in captive assurance colonies. In 2006, the Memphis Zoo (MZ) initiated a program to develop ART for the critically endangered Mississippi gopher frog after natural breeding failed. Standard gamete collection and IVF developed by MZ for reproducing endangered toads such as the Wyoming or boreal toad were applied to the gopher frog with little success, especially hormonal therapy for sperm production. Using the leopard frog as a model species for Ranids, we tested the time and dose dependence of a luteinizing hormone releasing hormone analogue (LHRHa) and hCG on sperm quantity and quality. Initial findings from the leopard frog study were critical in designing the study on gopher frogs. Our objectives were to (1) compare 2 different hormones administered intraperitoneal (500 IU hCG vs 15 μg LHRHa) or their combination on spermiation in gopher frogs; (2) develop in vivo oocyte maturation and ovulation protocols using LHRHa (15 μg) and hCG (500 IU); and (3) transfer this technology to another institution as proof of principle. In gopher frogs, 100 and 83% of the males produced sperm in response to the LHRHa and the combination treatment, respectively, whereas only 16% responded to hCG alone. Sperm concentration peaked at 1 h post-administration for all treatments, with the LHRH/hCG cocktail treatment producing the highest concentration of sperm (mean = 4.6 × 106 ± 1.2 × 106 sperm mL–1, n = 6). No differences in motility were observed between treatments (P > 0.05). For females, a series of priming hormones of hCG and LHRHa were given several months before an ovulatory hormone regimen resulting in ovulation by 100% of the females (n = 6), whereas animals not primed failed to ovulate (n = 4). These 3 separate priming and IVF trials conducted between 2008 and 2010 resulted in each female laying ∼2000 eggs, with an average fertilization rate of 76% for inseminated eggs and hundreds of tadpoles produced. These IVF tadpoles represent the first captive reproduction of gopher frogs and highlight how ART can be applied to conservation and genetic management of threatened species. Subsequently, we tested our IVF protocols on gopher frogs at Omaha's Henry Doorly Zoo using fresh (collected on site) and chilled, shipped sperm from MZ. We collected 6169 eggs from 9 hormone-primed females with all animals ovulating. A portion of the total eggs ovulated were inseminated, resulting in 2401 fertilized eggs (38.9% of total eggs collected) across 18 different male–female pairings leading to viable tadpoles. In addition, sperm transferred overnight from the MZ produced 202/441 fertilized eggs (46%). The transfer of this technology and production of endangered amphibians using chilled, shipped sperm from live animals is a conservation milestone that can be applied to other captive breeding programs.

Law in Ensuring the Safety of Genetic Technologies Against their Use for Criminal Purposes

Lex Russica ◽

10.17803/1729-5920.2020.165.8.047-053 ◽

2020 ◽

pp. 47-53

Author(s):

N. V. Kruchinina

Keyword(s):

Legal Status ◽

Legal Framework ◽

Legal Regulation ◽

Human Reproduction ◽

Economic Progress ◽

Genetic Technologies ◽

Criminal Groups ◽

Definition Of

Genetic technologies offer wide prospects for socio-economic progress. At the same time, their application in practice could put at stake the interests of society, human rights and freedoms. Therefore, the development of genetic technologies requires its analysis from the standpoint of jurisprudence, thoughtful legislative regulation and protection from uncontrolled spread and criminal use. The paper analyzes different points of view on the use of genetic technologies. The author substantiates the necessity of proper legal regulation and security of the process of development of genetic technologies. The paper contains the results of the scientific research. The paper elucidates the problems related to the use of genetic technologies in the process of artificial human reproduction: imperfection of the legal framework (In particular, lack of the definition of the legal status of human embryo, lack of justification for the legality of its use for research and therapeutic purposes), the threat of the use of genetic technologies for criminal purposes. The author concludes that the use of genetic technologies for criminal purposes is especially dangerous because organized criminal groups focus their attention on genetic technologies. This gives rise to a special criminal situation that requires new approaches for effective counteraction. To this end, the priority is given to identification of crimes committed with the use of genetic technologies and analysis of the emerging practice of investigating this category of crimes. Failure to comply with standards, deviation from regulations and procedures imposed on medical care may result in harm to health or death also when the assisted reproductive technologies are used. The author has made some proposals to solve these problems with due regard to domestic and foreign experience in the use of genetic technologies in the field of human artificial reproduction (in particular, it is proposed to establish effective international cooperation in this area).

Culture Medium and Sex Drive Epigenetic Reprogramming in Preimplantation Bovine Embryos

International Journal of Molecular Sciences ◽

10.3390/ijms22126426 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6426

Author(s):

Sebastian Canovas ◽

Elena Ivanova ◽

Meriem Hamdi ◽

Fernando Perez-Sanz ◽

Dimitrios Rizos ◽

...

Keyword(s):

Dna Methylation ◽

Developmental Process ◽

Sexually Dimorphic ◽

Element Analysis ◽

Global Methylation ◽

The Impact

Assisted reproductive technologies impact transcriptome and epigenome of embryos and can result in long-term phenotypic consequences. Whole-genome DNA methylation profiles from individual bovine blastocysts in vivo- and in vitro-derived (using three sources of protein: reproductive fluids, blood serum and bovine serum albumin) were generated. The impact of in vitro culture on DNA methylation was analyzed, and sex-specific methylation differences at blastocyst stage were uncovered. In vivo embryos showed the highest levels of methylation (29.5%), close to those produced in vitro with serum, whilst embryos produced in vitro with reproductive fluids or albumin showed less global methylation (25–25.4%). During repetitive element analysis, the serum group was the most affected. DNA methylation differences between in vivo and in vitro groups were more frequent in the first intron than in CpGi in promoters. Moreover, hierarchical cluster analysis showed that sex produced a stronger bias in the results than embryo origin. For each group, distance between male and female embryos varied, with in vivo blastocyst showing a lesser distance. Between the sexually dimorphic methylated tiles, which were biased to X-chromosome, critical factors for reproduction, developmental process, cell proliferation and DNA methylation machinery were included. These results support the idea that blastocysts show sexually-dimorphic DNA methylation patterns, and the known picture about the blastocyst methylome should be reconsidered.

In vivo measurement of pH and CO2 levels in the uterus of sows through the estrous cycle and after insemination

Scientific Reports ◽

10.1038/s41598-021-82620-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Octavio López-Albors ◽

Pedro José Llamas-López ◽

Joaquín Ángel Ortuño ◽

Rafael Latorre ◽

Francisco Alberto García-Vázquez

Keyword(s):

Estrous Cycle ◽

Invasive Approach ◽

Vitro Fertilization ◽

Non Invasive ◽

Physiological Values

AbstractThe pH–CO2–HCO3− system is a ubiquitous biological regulator with important functional implications for reproduction. Knowledge of the physiological values of its components is relevant for reproductive biology and the optimization of Assisted Reproductive Technologies (ARTs). However, in situ measurements of these parameters in the uterus are scarce or null. This study describes a non-invasive method for in situ time-lapse recording of pH and CO2 within the uterus of non-anesthetized sows. Animals were at three different reproductive conditions, estrous with no insemination and two hours after insemination, and diestrous. From pH and CO2 data, HCO3− concentration was estimated. The non-invasive approach to the porcine uterus with novel optical probes allowed the obtaining of in situ physiological values of pH, CO2, and HCO3−. Variable oscillatory patterns of pH, CO2 and HCO3− were found independently of the estrous condition. Insemination did not immediately change the levels of uterine pH, CO2 (%) and HCO3− concentration, but all the values were affected by the estrous cycle decreasing significantly at diestrous condition. This study contributes to a better understanding of the in vivo regulation of the pH-CO2-HCO3− system in the uterus and may help to optimize the protocols of sperm treatment for in vitro fertilization.

HYPOXIA AND REPRODUCTIVE HEALTH: Hypoxia and ovarian function: follicle development, ovulation, oocyte maturation

Reproduction ◽

10.1530/rep-20-0509 ◽

2021 ◽

Vol 161 (1) ◽

pp. F33-F40

Author(s):

Megan Lim ◽

Jeremy G Thompson ◽

Kylie R Dunning

Keyword(s):

Oocyte Maturation ◽

Follicle Development ◽

Oxygen Regulation ◽

Low Oxygen ◽

Oxygen Environment ◽

Regulated Gene Expression

The ovarian follicle provides the oocyte with the ideal environment for growth and development in preparation for ovulation and fertilisation. The follicle undergoes many structural changes as it grows, including changes in vasculature, cell proliferation and differentiation and the formation of a fluid-filled antrum. These changes collectively create a low oxygen environment within the follicle. Thus, the oocyte itself develops in a potentially hypoxic environment. The survival of hypoxic tissues is controlled by hypoxia-inducible factors (HIFs) that are activated in a low oxygen state. The understanding of HIF pathways is growing across all fields of biology, and its role in ovarian development is steadily gaining clarity. One of the genes upregulated by HIF is a vascular endothelial growth factor, the main inducer of angiogenesis which is required for follicle development and corpus formation. Ovulation is also intrinsically linked to HIF activity through the ovulatory luteinising hormone surge increasing HIF expression. The role for HIF in oocyte maturation is less understood, as efforts to replicate the low oxygen environment of the in vivo follicle are not achievable by culturing in low oxygen alone. There is potential for other factors present in vivo, but lost in vitro, to be involved in oxygen regulation. One factor of interest is haemoglobin, the oxygen-binding protein, which brings the exciting possibility of sensitive oxygen regulation, consequently affecting HIF-regulated gene expression. A thorough understanding of oxygen regulation within the follicle would provide vital applications for the field of assisted reproductive technologies, in particular in vitro oocyte maturation.

196 EFFECTS OF DIFFERENT IN VITRO MATURATION SYSTEMS ON BOVINE EMBRYO DEVELOPMENT

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab196 ◽

2012 ◽

Vol 24 (1) ◽

pp. 210

Author(s):

S. M. Bernal ◽

J. Heinzmann ◽

D. Herrmann ◽

A. Lucas-Hahn ◽

B. Timmermann ◽

...

Keyword(s):

Mrna Expression ◽

In Vitro Maturation ◽

Expression Profiles ◽

Developmental Competence ◽

Bovine Embryo ◽

Blastocyst Quality

Bovine oocytes and embryos have been established as a valuable model for studying human early development, specifically after assisted reproductive technologies (ART). Efforts for the improvement of ART in the last years have focused on culture media and conditions. Recently, Albuz et al. (2010) reported that the culture of bovine cumulus–oocyte complexes (COC) with cyclic adenosine 3′, 5′-monophosphate (cAMP) modulators, before and during extended in vitro maturation (IVM), improved blastocyst quality and yields in mice and cattle. In this study, we investigated the influence of an extended IVM phase in combination with cAMP modulators on blastocyst yields and quality, the effects on mRNA expression profiles and epigenetic marks. We compared these results to the standard protocol (Wrenzycki et al., 2001) used in our laboratory with oocytes from different retrieval methods. Oocytes were retrieved from slaughterhouse ovaries either by slicing or follicular aspiration. The COC were either subjected directly to IVM using the standard TCM-based protocol for 24 h (TCM24-slicing and TCM24-aspiration, respectively) or oocytes that were retrieved by aspiration were treated with forskolin and IBMX for a 2-h pre-IVM period, followed by an extended IVM phase of 30 h in TCM, supplemented with cilostamide (cAMP30-aspiration). Statistical analyses were performed using 1-way ANOVA followed by the nonparametrical Kruskal–Wallis test. Maturation rates were 79.3 ± 2.6% in TCM24-aspiration, 74.2 ± 8.8% in cAMP30-aspiration and 70.4 ± 5.1% in TCM24-slicing oocytes. Matured oocytes were fertilized in vitro with semen from a bull previously proven to be suitable for IVF. Blastocyst rates from presumptive zygotes were significantly higher (P = 0.003) in the TCM24-aspiration group (32 ± 7%) compared to TCM24-slicing (23 ± 7%) and cAMP30-aspiration (22 ± 5%). Analysis revealed that cell numbers were rather similar in the 3 experimental groups (125 ± 19, 128 ± 15 and 129 ± 9), while in vivo-produced blastocysts possessed slightly more cells (134 ± 17; P ≥ 0.05). RT-qPCR analysis of mRNA expression for a panel of genes indicative of embryo quality including DNMT3a, SLC2A8, COX2 and PCK2, showed that blastocysts derived from both aspiration protocols were similar to in vivo embryos, but were different from blastocysts resulting from the ovary-slicing protocol. Specifically, the expression profile of COX2, which is involved in pregnancy outcome and in the response to growth factors, indicates an enhanced developmental competence of aspirated oocytes. However, the transcript level of EGR1 (early growth response) was significantly higher (P = 0.009) in in vivo-derived blastocysts in comparison to all in vitro treatments. The investigation of the epigenetic status of the in vitro-derived blastocysts based on bisulfite sequencing of 2 satellite repeat sequences is currently underway. Results so far indicate that the method of obtaining the oocytes (slicing vs aspiration) for in vitro production of bovine embryos is of greater influence on blastocyst quality than IVM conditions.