Evaluating risk, safety and efficacy of novel reproductive techniques and therapies through the EuroGTP II risk assessment tool.

STUDY QUESTION: Can risks associated with novelties in assisted reproduction technologies (ARTs) be assessed in a systematic and structured way? SUMMARY ANSWER: An ART-specific risk assessment tool has been developed to assess the risks associated with the development of novelties in ART (EuroGTP II-ART). WHAT IS KNOWN ALREADY: How to implement new technologies in ART is well-described in the literature. The successive steps should include testing in animal models, executing pre-clinical studies using supernumerary gametes or embryos, prospective clinical trials and finally, short- and long-term follow-up studies on the health of the offspring. A framework categorizing treatments from experimental through innovative to established according to the extent of the studies conducted has been devised. However, a systematic and standardized methodology to facilitate risk evaluation before innovations are performed in a clinical setting is lacking. STUDY DESIGN, SIZE, DURATION: The EuroGTP II-ART risk assessment tool was developed on the basis of a generic risk assessment algorithm developed for tissue and cell therapies and products (TCTPs) in the context of the project 'Good Practices for demonstrating safety and quality through recipient follow-up European Good Tissue and cells Practices II (EuroGTP II)'. For this purpose, a series of four meetings was held in which eight ART experts participated. In addition, several tests and simulations were undertaken to fine-tune the final tool. PARTICIPANTS/MATERIALS, SETTING, METHODS: The three steps comprising the EuroGTP II methodology were evaluated against its usefulness and applicability in ART. Ways to improve and adapt the methodology into ART risk assessment were agreed and implemented. MAIN RESULTS AND THE ROLE OF CHANCE: Assessment of the novelty (Step 1), consisting of seven questions, is the same as for other TCTPs. Practical examples were included for better understanding. Identification of potential risks and consequences (Step 2), consisting of a series of risks and risk consequences to consider during risk assessment, was adapted from the generic methodology, adding more potential risks for processes involving gonadic tissues. The algorithm to score risks was also adapted, giving a specific range of highest possible risk scores. A list of strategies for risk reduction and definition of extended studies required to ensure effectiveness and safety (Step 3) was also produced by the ART experts, based on generic EuroGTP II methodology. Several explanations and examples were provided for each of the steps for better understanding within this field. LIMITATIONS, REASONS FOR CAUTION: A multidisciplinary team is needed to perform risk assessment, to interpret results and to determine risk mitigation strategies and/or next steps required to ensure the safety in the clinical use of novelties. WIDER IMPLICATIONS OF THE FINDINGS: This is a dynamic tool whose value goes beyond assessment of risk before implementing a novel ART in clinical practice, to re-evaluate risks based on information collected during the process. STUDY FUNDING / COMPETING INTEREST(S): This study was called EUROGTP II and was funded by the European Commission (Grant agreement number 709567). The authors declare no competing interests concerning the results of this study.

Melatonin Receptor 1B and Corticosteroid Receptor Polymorphisms in Infertile Women with Implantation Failure and Miscarriages.

BACKGROUND: The development of assisted reproductive techniques has significantly improved fertility chances in many women, but recurrent implantation failure (RIF) and miscarriages (RM) might preclude successful pregnancy. Alterations in the intrinsic secretory patterns of melatonin and cortisol influence reproduction in humans, and imperfection of receptor - dependent signaling may additionally compromise the hormonal effects. Therefore, the present study aims to investigate the influence of certain melatonin and cortisol receptor polymorphisms in infertile women. METHODS: A total of 111 female infertile patients suffering from implantation failure and/or miscarriages were genotyped for MTNR1B rs1562444, MTNR1Brs10830962, GCCR rs41423247, and GCCR ER22/23EK variants. Additionally, 106 female volunteers were genotyped for the same polymorphisms. RESULTS: The allele and genotype distribution of the investigated polymorphisms did not differ between infertile women and the control group. Significantly more women with history of RIF have MTNR1B rs1562444 G-allele-containing genotypes in comparison to AA carriers (19.3% vs. 3.6%, p = 0.004). The minor allele of the ER22/23EK variant was more frequent in infertile patients with three or more unsuccessful implantation attempts than in other women (12.5% vs. 2.4%, p = 0.025). CONCLUSIONS: Melatonin receptor 1B polymorphisms might affect embryo implantation and early pregnancy loss, while their influence on late pregnancy complications needs further evaluation. The possible association between the cortisol receptor ER22/23EK variant and recurrent implantation failure might help to differentiate women who could benefit from corticosteroid treatment.

Decreased ratio of FOXP3+/FOXP3-CD45RA+CD4+ T cells in peripheral blood is associated with unexplained infertility and ART failure.

Unexplained infertility has a huge social impact and is a significant challenge for both clinicians and researchers. Previous studies have shown the involvement of multiple factors in infertility. Among these, the subset of regulatory T cells is of particular interest for the maternal tolerance towards the semi-allogenic fetus. We investigated circulating CD45RA+ regulatory and non-regulatory CD4+ T cells in healthy women and patients with unexplained infertility in the context of thymic output and peripheral proliferation. The proportion of FOXP3+ and FOXP3-CD45RA+CD4+ T cells in peripheral blood was studied in control groups of healthy parous and nulliparous (never-pregnant) women and in patients with unexplained infertility. In the same groups thymic output and peripheral proliferation were defined by the sj/ßTREC ratio, and signal joint T-cell receptor excision circles (sjTREC) and Ki67 expression, respectively. In parous women a decrease in sjTREC/105 cells and CD45RA+ T lymphocytes, compared to nulliparous group was found. At the same time, the proportion of FOXP3-CD45RA+CD4+ cells, but not FOXP3+CD45RA+ Tregs was reduced. In contrast, in patients with unsuccessful pregnancy, proportions of both regulatory and non-regulatory T cell counterparts were lower. Taken together, our results provide evidence for group-specific properties in the CD45RA+ T cell compartment between healthy parous, nulliparous and women with unexplained infertility.

Valor de la determinación del fenotipo del sistema Pi en la deficiencia de alfa-L-antitripsina (AL-AT) / Value of determination of Pi phenotype in alpha-L-antitrypsin deficiency (AL-AT)

Se estudian 110 niños con diversas enfermedades del hígado: 16 hepatitis agudas virales, 8 hepatitis crónicas persistentes, 31 hepatitis crónica activas, 5 esteatosis hepáticas, 5 cirrosis hepáticas, 24 colestasis del lactante, 3 enfermedad de Wilson, 2 fibrosis hepáticas congénitas, 5 enfermedades metabólicas y 5 de otras causas, con edades comprendidas entre 2 meses y 14 años, a los que se determinó el fenotipo del sistema Pi por enfoque isoelétrico en geles ultrafinos de poliacrilamida según el método de Kueppers, con modificaciones como pesquisaje de deficiencia de alfa-L-antitripsina (AL-AT), realizándose los niveles en suero a los que presentaban el fenotipo Pi ZZ, biopsia hepática con coloración de PAS con digestión de diastasa y estudio familiar del fenotipo. Se encuentra el 3,6% de positividad del fenotipo Pi ZZ en los niños estudiados, presentando disminución sérica de AL-AT y la presencia de inclusiones PAS positivas resistentes a diastasa en el citoplasma de los hepatocitos, teniendo 3 de ellos antecedentes de ictericia en la etapa posnatal y el cuarto paciente hepatomegalia como forma de presentación. Se observa que el estudio del fenotipo a los padres resultó un patrón heterocigótico (MZ) y los hermanos normales (MM). Se insiste en la importancia del diagnóstico de la deficiencia de AL-AT y el valor diagnóstico de la determinación del fenotipo del sistema Pi en toda enfermedad hepática en la infancia y adolescencia

La deficiencia de alfa 1-antitripsina como causa de enfermedad hepática crónica en la infancia / Alpha 1-antitrypsin deficiency as cause of chronic hepatic disease during infancy

Se presentan 4 pacientes con deficiencias de alfa-1-antitripsina (alfa-1-At), basado en diagnóstico clínico, bioquímico, laparoscopia y estudio histomorfológico del hígado; 3 de ellos con antecedentes de colestasis neonatal, con diagnóstico de hepatitis crónica activa el primero; el segundo evolucionó hacia una hipertensión portal precoz con cirrosis hepática y muerte a los 2 años y medio, y el tercero hacia una recuperación clínica, emdoscópica e hística. El cuarto paciente con hepatomegalia como forma de presentación y diagnóstico de hepatitis crónica activa. Todos los pacientes tenían disminución sérica de alfa-1-At, presencia de inclusiones ácido peryódico de Schiff (PAS) positivas resistentes a la diastasa en el citoplasma de los hepatocitos y fenotipo del sistema Pi ZZ. El estudio familiar del fenotipo de los padres resultó un patrón heterozigótico (MZ) y los hermanos normales (MM). Se insiste en el diagnóstico de esta enfermedad en todo niño con colestasis neonatal o enfermedad hepática antes de los 5 años de edad