ABSTRACT
Objective: This study analyzed the relation between the position of scientists on embryo editing and the different types of knowledge involved. Methods: A lexical analysis of 151 scientific articles in the PubMed and Web of Science databases was conducted using the IRAMUTEQ software. Results: The results showed that gene editing in embryos is presented in two argumentative branches: the first refers to the editing technique and its possibilities; the second discusses the impacts of these techniques on the public arena. The results demonstrate a consensus regarding the potential of editing; however, dilemmas about its effectiveness were also highlighted. Conclusion: The presence of ethical conflicts with embryo editing among the specialists was evidenced especially regarding the birth of genetically modified babies. Therefore, gene editing is marked by conflicts that are not limited only to biological contexts, but that encompasses different aspects of social life.
Objetivo: O objetivo deste trabalho foi analisar a relação entre o posicionamento dos cientistas sobre a edição de embriões e os diferentes tipos de conhecimento envolvidos nesses debates. Método: Utilizando o software IRAMUTEQ realizou-se uma análise lexical de 151 artigos científicos nas bases de dados PubMed e Web of Science. Resultados: Os resultados demonstraram que a edição genética de embriões se apresenta em dois blocos argumentativos: o primeiro se refere à técnica de edição e suas possibilidades e o segundo discute os impactos dessas técnicas na arena pública. Os achados demonstram consenso sobre as potencialidades da edição, contudo dilemas sobre a sua eficácia foram também destacados. Conclusão: Evidenciou-se a presença de embates éticos sobre a edição de embriões entre os especialistas em relação ao nascimento de bebês geneticamente modificados. Observou-se que a edição genética é marcada por conflitos que não se limitam apenas a contextos biológicos, mas que tangem diferentes aspectos da vida social.
Subject(s)
Bioethics , Embryo, Mammalian , Gene Editing , Social RepresentationABSTRACT
Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Subject(s)
Humans , Apolipoprotein E4/genetics , Cytosine , Mutation , Blastocyst , Heterozygote , Gene Editing , CRISPR-Cas SystemsABSTRACT
Introducción: Pese al incremento en conocimientos de la morfogénesis cardiaca humana, se conoce poco sobre los detalles cuantitativos en ello. Objetivo: Describir cuantitativamente el desarrollo del miocardio ventricular compacto y no compacto y su correlación con la longitud cráneo rabadilla. Métodos: Estudio descriptivo, transversal en 18 embriones humanos de los estadios 17 al 23 de Carnegie, pertenecientes a la Embrioteca de la Universidad de Ciencias Médicas de Villa Clara. Se midió la longitud del embrión, el grosor del miocardio compacto, trabecular y total en la pared lateral de ambos ventrículos y del vértice cardiaco. Resultados: El grosor de la pared lateral del miocardio compacto aumenta en ambos ventrículos desde los estadios 17 al 23 de Carnegie, de 0,06 mm hasta 0,17 mm en el derecho y de 0,09 mm hasta 0,23 mm en el izquierdo. El grosor de la pared lateral trabeculada disminuye con el avance de los estadios, de 0,43 mm a 0,34 mm en el derecho y de 0,45 mm a 0,37 mm en el izquierdo. El grosor de la pared lateral total aumenta de 0,48 mm a 0,51 mm en el ventrículo derecho y de 0,52 mm a 0,62 mm en el izquierdo. El grosor de la pared del vértice compacto aumenta de 0,19 mm a 0,25 mm. Conclusiones: La compactación de la pared ventricular aumenta con el desarrollo; la longitud cráneo raquis se relaciona con el grosor del miocardio ventricular(AU)
Introduction: Despite the increase in knowledge of human morphogenesis, especially cardiogenesis and the processes by which the morphology of the ventricular myocardium is defined, little is known about the quantitative details in it. Objectives: To quantitatively describe the development of compact and non-compact ventricular myocardium and its correlation with cranio-rump length. Methods: descriptive, cross-sectional study in 18 human embryos from Carnegie stages 17 to 23, belonging to the Embryoteca of the Villa Clara University of Medical Sciences. The length of the embryo, the thickness of the compact, trabecular and total myocardium were measured in the lateral wall of both ventricles and the cardiac apex. Results: The thickness of the lateral wall of the compact myocardium increases in both ventricles from Carnegie stages 17 to 23, from 0.06 mm to 0.17 mm in the right and from 0.09 to 0.23 mm in the left ventricles. The thickness of the trabeculated lateral wall decreases with the advancement of the stages, from 0.43 mm to 0.34 mm in the right and from 0.45 mm to 0.37 mm in the left. The total lateral wall thickness increases from 0.48 mm to 0.51 mm in the right ventricle and from 0.52 mm to 0.62 mm in the left. The wall thickness of the compact vertex increases from 0.19 mm to 0.25 mm. Conclusions: Ventricular wall compaction increases with development; the cranio-spinal length is related to the thickness of the ventricular myocardium(AU)
Subject(s)
Humans , Embryonic Structures/embryology , Heart Ventricles/embryology , Morphogenesis/physiology , Epidemiology, Descriptive , Cross-Sectional StudiesABSTRACT
RESUMEN Introducción: Aún persisten controversias en los eventos de la morfogénesis cardiovascular y una ausencia, casi total, de parámetros morfométricos en las fases iniciales de su desarrollo. Objetivos: Determinar la razón miocardio no compactado/miocardio compactado (NC/C) en ambos ventrículos y la evolución cronológica de esta razón en el período estudiado. Método: Se realizó un estudio descriptivo, transversal, con 18 embriones humanos pertenecientes a la Embrioteca de la Universidad de Ciencias Médicas de Villa Clara (Cuba) clasificados entre los estadios 17 y 23 de Carnegie. Se determinó el índice NC/C, el cual no es más que el cálculo matemático de la razón entre las porciones no compactada y compactada por espécimen y por estadios. Resultados: Los resultados de la aplicación de este índice en el ventrículo derecho de los embriones son: 7,17; 4,26; 3,12; 2,79; 2,36; 2,84 y 2,10 en los estadios de Carnegie 17, 18, 19, 20, 21, 22 y 23, respectivamente. En estos mismos especímenes se obtuvo como resultado en el ventrículo izquierdo: 5,0; 3,80; 2,68; 2,18; 2,50; 2,01 y 1,56, igualmente organizado por estadios. Conclusiones: Los índices NC/C obtenidos sustentan cuantitativamente que la compactación del miocardio ventricular avanza en los estadios evaluados; sus valores, mayores en el vértice, denotan que es posible que aún no haya concluido en esta zona.
ABSTRACT Introduction: Controversies still persist regarding the events of cardiovascular morphogenesis and an almost total absence of morphometric parameters in the initial phases of its development. Objectives: To determine the non-compacted to compacted (NC/C) myocardium ratio in both ventricles and the chronological progression of this ratio in the period studied. Method: A descriptive, cross-sectional study was carried out with 18 human embryos belonging to the Embryoteca of the Universidad de Ciencias Médicas de Villa Clara (Cuba) classified between Carnegie stages 17 and 23. The NC/C ratio -which is simply the mathematical calculation of the ratio between the non-compacted and compacted portions per specimen and per stage- was determined. Results: The application of this ratio in the right ventricle of the embryos obtained the following results: 7.17; 4.26; 3.12; 2.79; 2.36; 2.84 and 2.10 in Carnegie's stages 17, 18, 19, 20, 21, 22 and 23, respectively. In these same specimens, the left ventricle yielded the following results: 5.0; 3.80; 2.68; 2.18; 2.50; 2.01 and 1.56, also organized by stages. Conclusions: NC/C ratios obtained quantitatively support a progression of the ventricular myocardial compaction in the evaluated stages; their higher values at the apex denote that it may still be incomplete in this zone.
Subject(s)
Embryo Research , Embryonic and Fetal DevelopmentABSTRACT
OBJECTIVE: It is widely accepted that aging decreases women’s fertility capacity. The aim of this study was to assess correlations between maternal age and the morphokinetic parameters and cleavage pattern of embryos. METHODS: The morphokinetics of embryos derived from women 40 years of age were compared retrospectively in terms of time of second polar body extrusion, time of pronuclei appearance, time of pronuclei fading, and time of two to eight discrete cells (t2–t8). Furthermore, abnormal cleavage patterns such as uneven blastomeres at the two-cell stage, cell fusion (Fu), and trichotomous mitoses (TM) were assessed. RESULTS: Only t5 occurred later in women aged 36–40 and >40 years when compared with those aged 0.05). However, Fu and TM were more common in women aged >40 years than in younger women (p<0.001). CONCLUSION: Maternal age was correlated with the cleavage pattern of embryos. Therefore, evaluating embryo morphokinetics may contribute to optimal embryo selection, thereby increasing fertility in patients with advanced maternal age.
Subject(s)
Female , Humans , Aging , Blastomeres , Cell Fusion , Embryonic Structures , Fertility , Maternal Age , Mitosis , Polar Bodies , Retrospective Studies , Sperm Injections, IntracytoplasmicABSTRACT
The human nervus terminalis (terminal nerve) and the nerves to the vomeronasal organ (VNON) are both associated with the olfactory nerves and are of major interest to embryologists. However, there is still limited knowledge on their topographical anatomy in the nasal septum and on the number and distribution of ganglion cells along and near the cribriform plate of the ethmoid bone. We observed serial or semiserial sections of 30 fetuses at 7–18 weeks (crown rump length [CRL], 25–160 mm). Calretinin and S100 protein staining demonstrated not only the terminal nerve along the anterior edge of the perpendicular lamina of the ethmoid, but also the VNON along the posterior edge of the lamina. The terminal nerve was composed of 1–2 nerve bundles that passed through the anterior end of the cribriform plate, whereas the VNON consisted of 2–3 bundles behind the olfactory nerves. The terminal nerve ran along and crossed the posterior side of the nasal branch of the anterior ethmoidal nerve. Multiple clusters of small ganglion cells were found on the lateral surfaces of the ethmoid's crista galli, which are likely the origin of both the terminal nerve and VNON. The ganglions along the crista galli were ball-like and 15–20 µm in diameter and, ranged from 40–153 in unilateral number according to our counting at 21-µm-interval except for one specimen (480 neurons; CRL, 137 mm). An effect of nerve degeneration with increasing age seemed to be masked by a remarkable individual difference.
Subject(s)
Humans , Calbindin 2 , Ethmoid Bone , Fetus , Ganglion Cysts , Individuality , Masks , Nasal Septum , Nerve Degeneration , Neurons , Olfactory Nerve , Vomeronasal OrganABSTRACT
As the number of frozen human embryos continues to rise daily, with numbers not expected to fall, an answer must be found to this dilemma. Four possible solutions have been suggested: a) thaw the embryos and allow them to perish; b) thaw them and donate them for biomedical research; c) thaw them and donate them in adoption; and d) leave them frozen indefinitely. This paper will evaluate the morality of these four possible solutions, particularly frozen human embryo adoption in the light of the Magisterium of the Catholic Church, which in its Instruction Dignitas Personae, appears to have opted to consider this practice as illicit. We also review the various stances of expert moralists in favour of or against frozen human embryo adoption, and we reflect on the extent to which the doctrine contained in Dignitas Personae can bind the moral conscience of the Catholic faithful. Finally, we make a personal evaluation of frozen human embryo adoption, in an attempt to find moral reasons that substantiate the negative opinion manifested by the Catholic Magisterium on this matter. In conclusion, we suggest that the moral assessment of frozen human embryo adoption as set forth in Dignitas Personae might not be considered as settled. Therefore, we are of the opinion that there is no impediment to further research on the moral foundation of this adoptive practice; however, insofar as this occurs, we believe that the best attitude of the Catholic faithful is to follow the moral criteria presented in Dignitas Personae with respect to the adoption of frozen human embryos.
Cada día aumenta el número de embriones humanos congelados y no se prevé que su número disminuya, por lo que parece necesario buscar una solución a este problema. Se han planteado cuatro posibles: a) descongelarlos y dejarlos morir; b) descongelarlos y donarlos para investigaciones biomédicas; c) descongelarlos y donarlos en adopción; y d) dejarlos congelados indefinidamente. En este trabajo se evalúa la moralidad de estas cuatro posibles soluciones, y espacialmente de la adopción de los embriones humanos congelados a la luz del Magisterio de la Iglesia Católica, que en su Instrucción Dignitas Personae, se decanta por la ilicitud de dicha práctica. También se revisan distintas posturas de moralistas expertos favorables o no a la adopción de embriones humanos congelados. Igualmente se reflexiona sobre en qué medida la doctrina contenida en Dignitas Personae puede obligar a la conciencia moral de los fieles católicos. Finalmente se realiza una evaluación personal de la adopción de embriones humanos congelados tratando de buscar razones morales que fundamenten el porqué del juicio negativo manifestado por el Magisterio Católico. Los autores sostienen que la ilicitud ética de la adopción de embriones humanos congelados puede radicar en la ilicitud moral de la subrogación, que hace ilícito todo el proceso procreativo, constituido por: acto conyugal, fecundación del óvulo e implantación del embrión producido en el útero en su madre biológica. Finalmente se plantea que lo expuesto en Dignitas Personae posiblemente no da por zanjada la valoración moral de la adopción de embriones humanos congelados, por lo que somos de la opinión de que no existe impedimento alguno para poder seguir investigando sobre la fundamentación moral de esta práctica adoptiva; pero que, en tanto en cuanto ello se produzca, nos parece que la mejor actitud de los fieles católicos es seguir los criterios morales de Dignitas Personae, expone con respecto a la adopción de embriones humanos congelados.
Cada dia aumenta o número de embriões humanos congelados e não se espera o número diminua, portanto parece necessário encontrar uma solução para este problema. Foram levantadas quatro possíveis: a) descongelá-los e deixá-los morrer; (b) descongelá-los e doá-los para pesquisa biomédica; (c) descongelada-los e doá-los para adoção; e (d) deixá-los congelados indefinidamente. Nesse trabalho se avalia a moralidade dessas quatro possíveis soluções e especialmente a adoção de embriões humanos congelados à luz do Magistério da Igreja Católica, que em sua instrução Dignitas Personae, opta pela ilegalidade da prática. Também se revisam posturas diferentes dos moralistas especialistas favoráveis ou não à adoção de embriões humanos congelados. Igualmente, reflete-se sobre como a doutrina contida na Dignitas Personae pode obrigar a consciência moral dos fiéis católicos.Finalmente se realiza uma avaliação pessoal da adoção de embriões humanos congelados pretendendo buscar razões morais que fundamentem o porquê do juízo negativo manifestado pelo magistério católico. Os autores argumentam que a ilegalidade ética da adoção de embriões humanos congelados pode resultar na ilicitude moral de sub-rogação, tornando ilícito todo o processoprocriador, constiuído por: ato conjugal, fertilização do óvulo e implantação do embrião produzido no útero de sua mãe biológica. Finalmente, apresenta-se que o exposto no Dignitas Personae, possivelmente, não dá por resolvida a valoração moral da adopção de embriões humanos congelados, portanto somos da opinião de que não há nenhum impedimento para seguir pesquisando sobre o fundamentação moral desta prática adotiva; Porém, porquanto ele se produza, parece-nos que a melhor atitude dos fiéis católicos será seguir os critérios morais de Dignitas Personae, expostos no que se refere a adoção de embriões humanos congelados.
Subject(s)
Humans , Adoption , Catholicism , Cryopreservation , Embryonic Structures , MoraleABSTRACT
Resumo: As indagações com relação ao termo inicial da vida humana perpassam a sociedade desde a antiguidade. No mundo pós-moderno, os avanços da ciência e da técnica potencializam as discussões acerca do tema, de modo que os debates que anteriormente se concentravam em torno do aborto, também passaram a ter como objeto as intervenções da biotecnologia. O presente artigo trata do último aspecto, a saber, refletir em que medida a dignidade humana pode ser considerada um referencial (hermenêutico) na construção de parâmetros ético-jurídicos para os avanços biotecnológicos na definição do marco inicial da vida humana. Para desenvolver a pesquisa, utilizou-se como método de abordagem a ética hermenêutica crítica, mediante a qual a ética encontra-se no centro do processo de compreensão e interpretação, observando-se os contornos da facticidade. Constatou-se que não há consenso no que diz respeito ao termo inicial da vida, de forma que é fundamental dialogar com a nova realidade decorrente dos avanços biotecnológicos, no processo de construção de preceitos éticos e jurídicos de proteção do embrião e da natureza humana, tendo como referencial a dignidade humana.
Abstract: Questions concerning the beginning of human life have pervaded society since antiquity. In the post-modern world, scientific and technological advances have fueled discussions on the issue, such that debates previously concentrated on abortion now also focus on biotechnological interventions. The article addresses the latter, reflecting on the extent to which human dignity can be considered a (hermeneutic) reference in establishing ethical and legal parameters for biotechnological advances in the definition of the beginning of human life. The study's method was critical hermeneutic ethics, with ethics at the center of the process of understanding and interpretation, observing the contours of facticity. No consensus was found on the beginning of human life, so it is essential to engage in dialogue with the new reality resulting from biotechnological advances in the process of defining ethical and legal principles for protecting the embryo and human nature, with human dignity as the reference.
Resumen: Las investigaciones relacionadas con el inicio de la vida humana han acompañado a la sociedad desde la antigüedad. En el mundo posmoderno, los avances de la ciencia y de la técnica potencian las discusiones sobre este tema, de modo que los debates que anteriormente se concentraban en torno al aborto, también pasaron a ser objeto por las intervenciones de la biotecnología. El presente artículo trata acerca de este último aspecto, a saber, reflexionar en qué medida la dignidad humana puede ser considerada una referencia (hermenéutica) en la construcción de parámetros ético-jurídicos para los avances biotecnológicos, en la definición del marco inicial de la vida humana. Para desarrollar la investigación, se utilizó como método de enfoque la ética hermenéutica crítica, mediante la cual la ética se encuentra en el centro del proceso de comprensión e interpretación, observándose los contornos de la facticidad. Se constató que no existe consenso en lo que se refiere al término inicial de la vida, de forma que es fundamental dialogar con la nueva realidad, derivada de los avances biotecnológicos, en el proceso de construcción de preceptos éticos y jurídicos de protección del embrión y de la naturaleza humana, teniendo como referente la dignidad humana.
Subject(s)
Humans , Biotechnology/ethics , Reproductive Techniques/ethics , Beginning of Human Life/ethics , Reproductive Health Services/ethics , Ethics, Medical , Biotechnology/legislation & jurisprudence , Reproductive Techniques/legislation & jurisprudence , Reproductive Health Services/legislation & jurisprudenceABSTRACT
β-Thalassemia is a global health issue, caused by mutations in the HBB gene. Among these mutations, HBB -28 (A>G) mutations is one of the three most common mutations in China and Southeast Asia patients with β-thalassemia. Correcting this mutation in human embryos may prevent the disease being passed onto future generations and cure anemia. Here we report the first study using base editor (BE) system to correct disease mutant in human embryos. Firstly, we produced a 293T cell line with an exogenous HBB -28 (A>G) mutant fragment for gRNAs and targeting efficiency evaluation. Then we collected primary skin fibroblast cells from a β-thalassemia patient with HBB -28 (A>G) homozygous mutation. Data showed that base editor could precisely correct HBB -28 (A>G) mutation in the patient's primary cells. To model homozygous mutation disease embryos, we constructed nuclear transfer embryos by fusing the lymphocyte or skin fibroblast cells with enucleated in vitro matured (IVM) oocytes. Notably, the gene correction efficiency was over 23.0% in these embryos by base editor. Although these embryos were still mosaic, the percentage of repaired blastomeres was over 20.0%. In addition, we found that base editor variants, with narrowed deamination window, could promote G-to-A conversion at HBB -28 site precisely in human embryos. Collectively, this study demonstrated the feasibility of curing genetic disease in human somatic cells and embryos by base editor system.
Subject(s)
Female , Humans , APOBEC-1 Deaminase , Genetics , Metabolism , Base Sequence , Blastomeres , Cell Biology , Metabolism , CRISPR-Cas Systems , Embryo, Mammalian , Metabolism , Pathology , Fibroblasts , Metabolism , Pathology , Gene Editing , Methods , Gene Expression , HEK293 Cells , Heterozygote , Homozygote , Point Mutation , Primary Cell Culture , Promoter Regions, Genetic , Sequence Analysis, DNA , beta-Globins , Genetics , Metabolism , beta-Thalassemia , Genetics , Metabolism , Pathology , TherapeuticsABSTRACT
Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
ABSTRACT
Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
ABSTRACT
La búsqueda de la eficacia en la fecundación in vitro hace que se produzcan más embriones que los que se implantarán, lo que produce un excedente de embriones, que es congelado. Esto hace que ineludiblemente el número de embriones humanos congelados aumente. Entre las soluciones para dichos embriones humanos congelados está la donación/adopción de los mismos. Ineludiblemente esta práctica conlleva objetivos problemas éticos. En este trabajo se evalúa la eticidad de la donación/adopción de embriones humanos congelados desde la perspectiva de la filosofía moral, lo que podríamos llamar una "ética laica" y dos de las religiones monoteístas: la musulmana y la judía.
The search for IVF efficacy leads to a higher embryo production than it is necessary for implantation; this results in an excess of embryos which are kept frozen. This amount of frozen embryos inevitably increases. The donation/adoption are among the possible solutions for these frozen embryos. However, this practice has objective ethical problems. This article considers the ethical aspects of the donation / adoption of frozen human embryos from the point of view of moral philosophy, from what we could call "secular ethics" and from two monotheistic religions: Muslim and Jewish.
A busca da eficácia na fecundação in vitro faz com que se produzam mais embriões dos que se implantarão, o que produz um excedente de embriões, que é congelado. Isto faz com que inquestionavelmente o número de embriões humanos congelados aumente. Entre as soluções para os ditos embriões humanos congelados está na doação/adoção dos mesmos. Ineludivelmente esta prática implica objetivos problemas éticos. Neste trabalho se avalia a eticidade da doação/adoção de embriões humanos congelados a partir da perspectiva da filosofia moral, o que poderíamos chamar uma "ética laica" e duas religiões monoteistas: a mulçumana e a judia.
Subject(s)
Cryopreservation , Embryo Culture Techniques/methods , Embryo Research/ethics , Embryo, Mammalian , Embryo Culture Techniques/ethics , Morale , ReligionABSTRACT
La utilización de embriones humanos en la investigación biológica ha generado un debate ético desde la discusión ontológica, en relación a si se puede o no considerar al embrión persona humana. En este artículo se análiza el estatuto ontológico del embrión humano desde una perspectiva biológica, considerando las principales líneas de investigación que lo intervienen en América Latina. Adcionalmente, revisamos el aporte desde el estatuto legal del embrión humano en esta región y la postura desde la formalidad de la investigación cientifica en el cuidado de su utilización. Consideramos finalmente que en América Latina no se ha abordado de manera profunda la discusión en torno al estatuto del embrión humano como persona, quedando la discusión en una dimensión biológica.
The utilization of human embryos in the biological research has generated a bioethical debate from the ontological point of view about whether or not to consider the embryo as a human person. This paper review the ontological statute of the human embryo from a biological perspective considering the principal lines of investigation in Latin America. Complementary to discussion we check the contribution from the legal statute of the human embryo and the position from the formal scientific research in his utilization. Finally, we consider that, in Latin America, the discussion has not been approached in a deep way about the statute of the human embryo as person, staying the discussion in a dimension of the biological thing.
A utilização de embriões humanos na investigação biológica tem gerado um debate ético a partir da discussão ontológica, com relação a se deveria ou não considerar o embrião pessoa humana. Neste artigo se analisa o estatuto ontológico do embrião humano a partir de uma perspectiva biológica, considerando as principais linhas de investigação de intervenção no embrião na América Latina. Adicionalmente, revisamos a contribuição a partir do estatuto legal do embrião humano nesta região e a postura a partir da formalidade da investigação cientifica no cuidado de sua utilização. Consideramos finalmente que na América Latina não se tem abordado de maneira profunda a discussão em torno do estatuto do embrião humano como pessoa, tornando-se a discussão uma dimensão biológica.
Subject(s)
Humans , Bioethics , Embryo Research/ethics , Embryo, Mammalian , Personhood , Embryo Research/legislation & jurisprudence , Latin AmericaABSTRACT
Objective To explore the effect of Dexamethasone(DEX)on the proliferation and the expression of fibroblast growth factor(FGF)- 1,FGF - 2 and interleukin - 6(IL - 6)mRNA in hyperoxia - exposed human em-bryo lung fibroblasts(MRC - 5). Methods High oxygen volume fraction of 950 mL/ L was used to establish hyperoxia -damaged cell models,then treated with different concentrations of DEX(10 - 4 ,10 - 6 ,10 - 8 ,10 - 9 ,10 - 10 ,10 - 11 ,10 - 12 mol/ L),respectively. The proliferation activity of the cells was evaluated by methyl thiazolyl tetrazolium(MTT)at 24 h,48 h,72 h,and the optimal hyperoxia - exposed time and concentration of DEX were chosen;then they were divided into air group,hyperoxia group and hyperoxia + DEX group. The mRNA expressions of FGF - 1,FGF - 2 and IL - 6 were detected by quantitative real - time PCR at 48 h. Results Cell proliferation activity of the hyperoxia group was lower than that of the air group and there were significant differences at 48 h and 72 h(all P ﹤ 0. 05). Compared with the hyperoxia group,cell proliferation activity of the hyperoxia + DEX group increased with the concentration of DEX decreasing,reaching the peak at 10 - 9 mol/ L,and then gradually decreased with the concentration of DEX decreasing. Cells were cultured for 48 h,compared with the air group,the level of FGF - 1 mRNA was lower,FGF - 2 and IL - 6 mRNA were higher in the hyperoxia group(P ﹤ 0. 05). Compared with hyperoxia group,the level of FGF - 1 mRNA was higher,FGF - 2 and IL - 6 mRNA were lower in hyperoxia + DEX group(P ﹤ 0. 05). Conclusions Exposure to high oxygen volume fraction of 950 mL/ L for 48 h can cultivate optimal hyperoxia - damaged cell models,and DEX can protect the cell from hyperoxia injury and 10 - 9 mol/ L was the optimal concentration. Enhanced the expression of FGF - 1 mRNA and inhibited expression of FGF - 2 mRNA may be one of the mechanism that DEX protects the cells from hyperoxia injury.
ABSTRACT
Objective To investigate the pathogen-related genes of atmospheric polluting disease so as to clarify the biology mechanism and provide the scientific basis.Methods By using the technique of dot blot hybridization,we analyzed genes’differential expression with cloning by exposure to ≥75 μg/m3 PM2.5 in heating season and < 75 μg/m3 PM2.5 in un-heating season in WI-38 human embryo lung cells.The levels of cytokines TNF-α,IL-2, IL-6 and IL-8 were determined by radio immunity assay. Results After 24h of treatment, compared with control group,more than 100μg/mL PM2.5 significantly increased TNF-α,IL-6 and IL-8 levels,and decreased IL-2 in WI-38 human embryo lung cells (P < 0.05 ).The clear stripe was found in 350 bp in 48 gene samples with segment with differential expression of genes exposed to different concentrations of PM2.5 in WI-38 human embryo lung cells.Through the dot blot hybridization,black brown spots were found in 41 samples in Tester cDNA hybridization,and no similar spots were found in all of the same samples in Driver cDNA hybridization. Conclusion PM2.5 exposure may induce the inflammatory damage of WI-38 human embryo lung cells.Obvious genetic damage was observed in those cells exposed to ≥75 μg/m3 PM2.5 in heating season.
ABSTRACT
In the embryonic heart, the primitive atrium is considered to receive the bilateral sinus horns including the upper terminal of the inferior vena cava (IVC). To reveal topographical anatomy of the embryonic venous pole of the heart, we examined horizontal serial paraffin sections of 15 human embryos with crown-rump length 9-31 mm, corresponding to a gestational age of 6-7 weeks or Carnegie stage 14-16. The IVC was often fixed to the developing right pulmonary vein by a mesentery-like fibrous tissue. Rather than the terminal portion of the future superior vena cava, the IVC contributed to form a right-sided atrial lumen at the stage. The sinus venosus or its left horn communicated with the IVC in earlier specimens, but in later specimens, the left atrium extended caudally to separate the sinus and IVC. In contrast, the right atrium consistently extended far caudally, even below the sinus horn, along the IVC. A small (or large) attachment between the left (or right) atrium and IVC in adult hearts seemed to be derived from the left (or right) sinus valve. This hypothesis did not contradict with the incorporation theory of the sinus valves into the atrial wall. Variations in topographical anatomy around the IVC, especially of the sinus valves, might not always depend on the stages but partly in individual differences.
Subject(s)
Adult , Animals , Humans , Crown-Rump Length , Embryonic Structures , Gestational Age , Heart , Heart Atria , Horns , Individuality , Paraffin , Pulmonary Veins , Vena Cava, Inferior , Vena Cava, SuperiorABSTRACT
A term "mesoesophagus" has been often used by surgeons, but the morphology was not described well. To better understand the structures attaching the human abdominal and lower thoracic esophagus to the body wall, we examined serial or semiserial sections from 10 embryos and 9 fetuses. The esophagus was initially embedded in a large posterior mesenchymal tissue, which included the vertebral column and aorta. Below the tracheal bifurcation at the fifth week, the esophagus formed a mesentery-like structure, which we call the "mesoesophagus," that was sculpted by the enlarging lungs and pleural cavity. The pneumatoenteric recess of the pleuroperitoneal canal was observed in the lowest part of the mesoesophagus. At the seventh week, the mesoesophagus was divided into the upper long and lower short parts by the diaphragm. Near the esophageal hiatus, the pleural cavity provided 1 or 2 recesses in the upper side, while the fetal adrenal gland in the left side was attached to the lower side of the mesoesophagus. At the 10th and 18th week, the mesoesophagus remained along the lower thoracic esophagus, but the abdominal esophagus attached to the diaphragm instead of to the left adrenal. The mesoesophagus did not contain any blood vessels from the aorta and to the azygos vein. The posterior attachment of the abdominal esophagus seemed to develop to the major part of the phrenoesophageal membrane with modification from the increased mass of the left fetal adrenal. After postnatal degeneration of the fetal adrenal, the abdominal esophagus might again obtain a mesentery. Consequently, the mesoesophagus seemed to correspond to a small area containing the pulmonary ligament and aorta in adults.
Subject(s)
Adult , Humans , Adrenal Glands , Aorta , Azygos Vein , Blood Vessels , Diaphragm , Embryonic Structures , Esophagus , Fetus , Ligaments , Lung , Membranes , Mesentery , Pleural Cavity , SpineABSTRACT
Analisam-se os argumentos usados pelas organizações de ética portuguesas na regulação da investigação em embriões de origem humana. Recolheram-se documentos produzidos entre 2006 e 2010. Procedeu-se à análise temática de conteúdo, e as estratégias discursivas foram estudadas a partir de uma abordagem semântica da informação. Discutiram-se o estatuto do embrião abstrato (ser humano/pessoa ou artefato biológico/neoestrutura laboratorial) e os critérios que devem nortear as boas práticas e equilibrar expectativas e riscos na investigação em embriões, coexistindo argumentos heterogéneos oriundos da bioética principialista, laica e interventiva. Importa incorporar no debate as perspetivas de quem tem que decidir o destino de embriões concretos.
This article analyzes the arguments used by Portuguese ethics organizations on to the regulation of human embryo research. Documents produced between 2006 and 2010 were collected and, based on thematic content analysis, the discursive strategies were studied from a semantic approach to data. The debate focused the status of abstract embryos (human being/person or biological artifact/laboratory neostructure) and the criteria that should guide best practices and balance expectations and risks on embryo research, in which heterogeneous arguments coexist based on principialist, secular and interventional bioethics. The perspectives of those who must decide the fate of real embryos should be incorporated into the discussion.
Subject(s)
Humans , Embryo, Mammalian , Ethics , Human Embryonic Stem Cells , PortugalABSTRACT
The role of bone morphogenetic proteins (BMP-s) in the development of the nervous system has been widely studied on avian and rodent embryos. Human embryos have rarely been available for detection of BMP expression. In this study 39 human embryos of Carnegie stages (CS) 10-20 were investigated. The embryos were fixed in paraformaldehyde, embedded in paraffin and sectioned serially in transverse direction. BMP-2 and BMP-4 protein expression in the developing neural tube and the caudal spinal cord was determined by immunohistochemistry. Our data show that BMP-s tend to be more expressed in the neural tube in earlier stages; in particular, BMP-4 staining was found to be higher at CS10 compared to CS20. More detailed analysis was performed on embryos of CS14-18. Stronger BMP-2 and BMP-4 expression was found in the dorsal part than in the ventral part of the spinal cord. No differences were seen in the staining intensity of BMP-s in the spinal ganglia. Interestingly, in neural crest cells BMP-2 staining was stronger at CS16 and CS18 as compared to CS14, while no differences were found in BMP-4 staining. On the other hand, in the non-neural ectoderm BMP-4 staining was found to be stronger at CS16 than at CS14, while no differences were seen for BMP-2. In conclusion, expression of BMP-s in the developing neural tube and spinal cord of human embryos is generally in accordance with the findings made in rodents and birds.
El papel de las proteínas morfogenéticas óseas (BMP-s) ha sido ampliamente estudiado en el desarrollo del sistema nervioso en embriones de aves y roedores. Los embriones humanos rara vez han estado disponibles para la detección de la expresión de BMP. En este estudio se investigaron 39 embriones humanos de los estadios Carnegie (CS) 10-20. Los embriones fueron fijados en paraformaldehído, embebidos en parafina y seccionados en serie en dirección transversal. Se determinó por inmunohistoquímica BMP-2 la expresión de la proteína BMP-4 en el tubo neural y en la médula espinal caudal en desarrollo. Nuestros resultados mostraron que la BMP-s tienden a ser más expresadas en el tubo neural en etapas tempranas, en particular, se encontró tinción BMP-4 más alta en comparación con CS10 CS20. Un análisis más detallado se realizó en embriones de CS14-18. En la parte dorsal se observó mayor expresión de BMP-2 y de BMP-4 que en la parte ventral de la médula espinal. No se observaron diferencias en la intensidad de la tinción de BMP-s en los ganglios espinales. Curiosamente, en las células de la cresta neural BMP-2 la tinción fue más fuerte en CS16 y CS18 en comparación con CS14, mientras que no se encontraron diferencias en la tinción de BMP-4. Por otro lado, en el ectodermo no neural se encontró tinción BMP-4 más fuerte en CS16 que en CS14, mientras que no se observaron diferencias para BMP-2. En conclusión, la expresión de BMP-s en el tubo neural en desarrollo y la médula espinal de embriones humanos está generalmente de acuerdo con los hallazgos realizados en roedores y aves.
Subject(s)
Humans , Spinal Cord/embryology , /physiology , /physiology , Neural Tube/embryology , Embryonic Development , ImmunohistochemistryABSTRACT
Este artigo trata da divergência científica existente com relação à sobra de embriões congelados, particularmente entre a biologia, a religião, o direito e a bioética, e focando especialmente nos meios e nas formas com que cada uma dessas ciências caracteriza o início da vida. É objetivo dos autores, ainda, fornecer alternativas para a proteção do embrião humano, como a adoção por casais ou apenas por mulheres que, por razões médicas, não podem ser fertilizadas, mas que podem gestar. Em se tratando de Brasil, os autores concluem ser legal e legítimo a doação e a adoção de embrião humano para fertilização, desde que sejam considerados o princípio da dignidade da pessoa humana e mantido o sigilo sobre a identidade dos doadores de gametas, cujos dados devem permanecer registrados de forma permanente no centro onde ocorreu a fertilização.
This article deals with the existing scientific divergence with regard to the surplus of frozen embryos, especially between biology, religion, the law and bioethics, with a specific focus on the ways in which each of these areas determines the onset of life. The aim of the authors is to suggest alternatives that protect the human embryo, such as adoption by couples or by single women, who, for medical reasons, are infertile, but are capable of bearing a child. In the case of Brazil, the authors conclude that it is legal and legitimate to donate and adopt human embryos for fertilization, so long as the principle of human dignity is upheld and confidentiality maintained regarding the identity of the donors of the gametes, whose data should remain on file permanently at the center where fertilization occurred.