Las contracciones en blastocistos humanos se correlacionan con aneuploidía, menor implantación y mayor tiempo de llegada a blastocisto: estudio retrospectivo con incubadora Time-Lapse

Objectives:To identify contraction (CT) patterns in human blastocysts using a Time-Lapse incubator and to correlate them with their ploidy status by PGT-A analysis, the time they took to reach blastocyst state, implantation rate and clinical pregnancy rate. Design: Retrospective cohort study. Interventions: From October 2016 through May 2018, 270 patients were evaluated, 912 embryos were cultivated in the Time-Lapse (Embryoscope™, Vitrolife) incubator; 778 of them were tested for aneuploidy using an NGS platform in a reference laboratory. Vitrification was performed according to the result of embryo development and awaiting the NGS result, followed by devitrification and transfer of a single embryo. Blastocyst contractions (CT) were determined using the EmbryoViewer (EmbryoViewer™ drawing tools) in order to obtain area, percentage of contraction and the various types of contractions, and these were compared with the NGS genetic study result. 182 transfers were performed to patients with an average age of 30.4 years, range 24 to 39 years. Finally, we obtained the correlation of implantation rate and clinical pregnancy of euploid embryos transferred in the program of assisted reproduction. Results: Embryos were separated into two groups according to contraction during their development: those that contracted (CT) and those that did not, named "only expanding" (OE). OE embryos were euploid in 58.3%, while 53.6% of CT embryos were aneuploid, with a statistically significant difference (p=0.029), meaning that OE embryos had higher chances of being euploid than CT embryos. Pregnancy rate was also higher in OE embryos (63.1% vs. 46.7% in CT embryos; p=0.012). CT embryos took longer to reach the blastocyst stage compared to OE embryos (p=0.004). Women's age had no correlation with embryo contraction. Conclusions: In this study, embryos who showed contractions had a higher chance of aneuploidy, a lower implantation rate, and required longer to reach the blastocyst stage. The simple fact of observing contractions in an embryo could be useful to decide to transfer another embryo without contractions. Further studies are necessary to prove these findings.

Objetivos. Estudiar los patrones de contracciones en blastocistos humanos mediante el uso de una incubadora time-lapse y correlacionarlos con su estado de ploidía por análisis PGT-A, el tiempo para alcanzar el estado blastocisto, la tasa de implantación y de embarazo clínico. Diseño. Estudio de cohortes retrospectivo. Intervenciones. Entre octubre 2016 y mayo 2018, se evaluó 270 pacientes; se hizo cultivo extendido de 5 a 6 días a 912 embriones en la incubadora time-lapse (Embryoscope, Vitrolife), y a 778 se les estudió para aneuploidía usando una plataforma NGS en un laboratorio de referencia. Hubo posterior vitrificación, según resultado del desarrollo embrionario y en espera del resultado del NGS, seguido de desvitrificación y transferencia de embrión único. Se determinó las contracciones del blastocisto (CTB) mediante la herramienta de dibujo del embrión EmbryoViewer (EmbryoViewer drawing tools), de manera de obtener el área, porcentaje de contracción y los diferentes tipos de contracciones, y se comparó los embriones con el resultado del estudio genético mediante NGS. Se transfirió 182 embriones en pacientes de 30,4 años promedio, rango entre 24 y 39 años. Finalmente, se correlacionó la tasa de implantación y embarazo clínico de los embriones euploides que fueron transferidos, en el programa de reproducción asistida. Resultados. Se separó los embriones en dos grupos de acuerdo a las contracciones durante su desarrollo, en aquellos que las tuvieron (CT) y aquellos que no, denominados 'solo expanding' (SE). Los embriones SE fueron euploides en 58,3%, mientras los embriones CT fueron aneuploides en 53,6%, con significancia estadística (p=0,012). Ello indica que la mayoría de los embriones euploides hacen 'solo expanding' durante su desarrollo, mientras que la mayoría de los embriones aneuploides (53,9%) hacen contracciones durante su desarrollo (p=0,029). Del mismo modo, la tasa de embarazo clínico de los embriones SE euploides fue 63,1% frente a 46,7% de los embriones CT, p=0.012. Finalmente, los embriones euploides CT tardaron más en convertirse en blastocistos tempranos que los embriones SE, p=0.004. La edad de la mujer no representó un factor para contracción embrionaria. Conclusiones. Los resultados obtenidos en este estudio muestran que los embriones que muestran contracciones, sin importar que tan intensas sean, están relacionados con mayor probabilidad de aneuploidías, menor tasa de implantación y ritmos de división lentos. El simple hecho de observar contracciones en un embrión podría ser útil para decidir transferir otro embrión que no las haya tenido. Se requiere más estudios para comprobar estos hallazgos.

El uso de la tecnología Time-Lapse y algoritmo predictivo KIDScore 5 ayuda a mejorar la selección de embriones euploides para la transferencia

Objectives: To determine if the use of the KIDScore 5 algorithm (known implantation data) can help select between euploid embryos in order to improve pregnancy and implantation rates in patients undergoing assisted reproductive procedures. Methods: Retrospective cohort study in a fertility clinic, from October 2016 to December 2018, of 1 049 embryos from 328 patients. All the embryos were cultured in the Time-Lapse, Embryoscope® incubator (Vitrolife®, Canada) for 5-6 days. Of these, 896 embryos (85.4%) were biopsied and analyzed by NGS, and assessed with the predictive KIDScore 5 algorithm (Vitrolife®, Canada). The 153 remaining embryos (14.6%) were assessed with the predictive KIDScore 5 algorithm only. 256 single euploid embryos were transferred in couples undergoing IVF treatments at the Inmater clinics laboratory of assisted reproduction in Lima - Peru. Results: The implantation rate was significantly higher (p = 0.004) in euploid embryos transferred when selected by the KIDScore 5 algorithm (Group 2) versus those selected using only genetic study by NGS technology (Group 1) (71.2% vs. 48.8%). The rate of implantation of the euploid embryos transferred with KIDScore value = 6 versus those transferred with KIDScore = 1 was statistically different (73.5% vs. 50.8%; p = 0.030). When assessing the relationship between the rate of euploid embryos versus the result of the KIDScore 5 value, we found highly significant differences in the rates of euploid embryos with values 6 and 5 versus those with KIDScore 0 and 1 (60.5% vs. 45.7%; p = 0.0004). Conclusions: The embryo selection with the KIDScore 5 algorithm offers advantage on implantation and pregnancy rates only when euploid embryos are transferred. Its use as an additional criterion to embryo selection should be considered when accompanied by genetic study of the embryos to be transferred. Euploid embryos with a higher value in the KIDScore 5 algorithm scale have better rates of implantation and euploidy than embryos with the minimum value of this algorithm.

Objetivos. Evaluar si el uso del algoritmo KIDScore 5 (known implantation data) puede ayudar a seleccionar entre los embriones euploides, para mejorar las tasas de embarazo e implantación en pacientes sometidas a procedimientos de reproducción asistida. Métodos. Estudio de cohorte retrospectivo en una clínica de fertilidad, desde octubre 2016 a diciembre 2018. Se estudió 1 049 embriones provenientes de 328 pacientes. Todos los embriones fueron cultivados en la incubadora Time-Lapse, Embryoscope® (Vitrolife®, Canadá) durante 5 a 6 días. De estos, 896 embriones (85,4%) fueron biopsiados y analizados mediante NGS y recibieron una valoración otorgada por el algoritmo predictivo KIDScore 5 (Vitrolife®, Canadá). Los 153 embriones restantes (14,6%) únicamente recibieron la valoración mediante el algoritmo predictivo KIDScore 5. Se realizó 256 transferencias únicas de embriones euploides en parejas sometidas a tratamientos de FIV en el laboratorio de reproducción asistida de la Clínica Inmater, Lima - Perú. Resultados. La tasa de implantación de los embriones euploides transferidos con valores de KIDScore = 6 versus los transferidos con valores de KIDScore = 1 tuvo diferencia estadísticamente significativa (73,5% vs. 50,8%; p=0,030). Al evaluar la relación entre la tasa de euploidia embrionaria versus el resultado del valor de KIDScore 5, se obtuvo diferencias altamente significativas en las tasas de euploidia en los embriones con resultados de KIDScore 6 y 5 versus los de KIDScore 0 y 1 (60,5% vs. 45,7%; p=0,0004). Conclusiones. La selección embrionaria con ayuda del algoritmo KIDScore 5 ofrece ventaja en las tasas de implantación y embarazo únicamente cuando se transfieren embriones euploides. Su uso como criterio adicional a la selección embrionaria debiera ser considerado siempre que se acompañe estudio genético a los embriones a transferir. Los embriones euploides con valor más alto en la escala del algoritmo KIDScore 5, tienen mejores tasas de implantación y euploidía que los embriones con el valor mínimo de dicho algoritmo.

Retrospective study of single vitrified-warmed blastocyst transfer cycles according to the presence of morphokinetic variables / 대한생식의학회지

This study retrospectively assessed whether time-lapse data relating to developmental timing and morphology were associated with clinical outcomes, with the eventual goal of using morphokinetic variables to select embryos prospectively for cryopreservation. In this study, we examined the clinical outcomes of single vitrified-warmed blastocyst transfer cycles that were cultured in a time-lapse incubation system. The morphokinetic variables included uneven pronuclei, an uneven blastomere, multinucleation, and direct, rapid, and irregular division. A total of 164 single vitrified-warmed blastocyst transfer cycles were analyzed (102 cycles of regularly developed blastocysts and 62 cycles of blastocysts with morphokinetic variables). No significant differences in the age of females or the standard blastocyst morphology were found between these two groups. The regularly developed blastocysts showed significantly higher implantation and clinical pregnancy rates than the blastocysts exhibiting morphokinetic variables (30.4% vs. 9.7% and 37.3% vs. 14.5%, respectively; p < 0.01). The blastocysts that exhibited morphokinetic variables showed different mean development times compared with the regularly developed blastocysts. Although morphokinetic variables are known to have fatal impacts on embryonic development, a considerable number of embryos developed to the blastocyst stage. Morphokinetic variables had negative effects on the implantation and clinical pregnancy rates in vitrified-warmed blastocyst transfer cycles. These findings suggest that blastocysts cultured in a time-lapse incubation system should be considered for selective cryopreservation according to morphokinetic variables.

Time lapse between endodontic and periodontal treatments of combined periodontal-endodontic lesion: a systematic review / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aims to evaluate the time lapse between the endodontic and periodontal treatments of the combined periodontal-endodontic lesion to guide the clinical treatment.</p><p><b>METHODS</b>A systemic literature search was performed in articles published from 1980 to March 2017 using the electronic databases, including PubMed, EMbase, Cochrane, Web of Science, CNKI, CBM, and Wanfang Databases.</p><p><b>RESULTS</b>After screening, two randomized controlled trials, two prospective case series studies, and eight case reports were ultimately included. All clinical studies and case reports were treated with root canal therapy and periodontal nonsurgical or periodontal surgical treatment, with root canal therapy as the initial treatment step. Changes in the periodontal probing depth and attachment loss were reported in all clinical studies and case reports.</p><p><b>CONCLUSIONS</b>An observational period between the endodontic and periodontal nonsurgical treatments may not be required. However, given that the quality of several included studies was limited, and the number of randomized controlled trials was small, a large number of randomized controlled trials are needed to verify this result.</p>

Research progress on new techniques for preimplantation genetic diagnosis / 医学研究生学报

Preimplantation genetic diagnosis (PGD) was developed to play a supporting role in assisted reproductive technology. With this kind of detection method, embryos with copy number variations, chromosome translocations or single mutations were excluded and the normal embryos were chosen and implanted. Theoretically, the application of these procedures could improve the implantation and pregnancy rate and help to delivery healthy offspring. PGD was considered to be more precise, higher specific and non-invasive with the appearance of microarray hybridization technology, the next generation sequencing and time-lapse monitoring technology. This paper presented a review of new Methods used in PGD, including fluorescence in situ hybridization, array comparative genomic hybridization, SNP array, next generation sequencing, MicroSeq-PGD, MaReCs, time-lapse monitoring and cfDNA-based method, and their advantages and disadvantages as well as efficacy in the procedures in which they are used.

Evaluation of human embryo development in in vitro fertilization- and intracytoplasmic sperm injection-fertilized oocytes: A time-lapse study / 대한생식의학회지

OBJECTIVE: We investigated whether the insemination method (in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI]) affected morphokinetic events and abnormal cleavage events in embryonic development. METHODS: A total of 1,830 normal fertilized embryos were obtained from 272 IVF and ICSI cycles that underwent ovum retrieval culture using a time-lapse system (Embryoscope) from June 2013 to March 2015. All embryos were investigated by a detailed time-lapse analysis that measured the developmental events in the hours after IVF or ICSI insemination. RESULTS: No significant differences were observed between the two groups regarding clinical outcomes (p>0.05). ICSI-derived embryos showed significantly faster morphokinetics than those derived from conventional IVF, from the time to pronuclear fading to the time to 6 cells (p0.05). There were no differences in abnormal cleavage events between the two groups (p>0.05); they showed the same rates of direct cleavage from 1 to 3 cells, 2 multinucleated cells, 2 uneven cells, and reverse cleavage. CONCLUSION: The morphokinetics of embryo development was found to vary between IVF- and ICSI-fertilized oocytes, at least until the 6-cell stage. However, these differences did not affect the clinical outcomes of the embryo. Additionally, no significant differences in abnormal cleavage events were found according to the fertilization method.

Mouse Fyn induces pseudopodium formation in Chinese hamster ovary cells

Molecular mechanisms underlying the effects of Fyn on cell morphology, pseudopodium movement, and cell migration were investigated. The Fyn gene was subcloned into pEGFP-N1 to produce pEGFP-N1-Fyn. Chinese hamster ovary (CHO) cells were transfected with pEGFP-N1-Fyn. The expression of Fyn mRNA and proteins was monitored by reverse transcription-PCR and Western blotting. Additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. Sequences of the recombinant plasmids pMD18-T-Fyn and pEGFP-N1-Fyn were confirmed by sequence identification using National Center for Biotechnology Information in USA, and Fyn expression was detected by RT-PCR and Western blotting. The morphology of CHO cells transfected with the recombinant vector was significantly altered. Fyn expression induced filopodia and lamellipodia formation. Based on these results, we concluded that overexpression of mouse Fyn induces the formation of filopodia and lamellipodia in CHO cells, and promotes cell movement.

A tecnologia time-lapse pode predizer a qualidade embrionária antes dos resultados do PGD (diagnósticogenético pré-implantacional)? / Technology time-lapse can predict embryo quality of results before PGD(pre-implantation genetic diagnosis)?

Objetivo: Verificar se há correlação entre os resultados obtidos via diagnóstico genético préimplantacional (PGD) e os dados obtidos via monitoramento em tempo real (time-lapse) durante os três dias de desenvolvimento embrionário. Método: Estudo retrospectivo no qual foram avaliados embriões de ciclos de injeção intracitoplasmática de espermatozóides (ICSI) de março a junho de 2012. Após a ICSI, os embriões foram colocados em incubadora vertical com monitoramento time-lapse durante três dias. A seguir os embriões foram submetidos ao PGD. Os resultados obtidos foram baseados na análise do tempo de aparecimento e desaparecimento dos dois pró-núcleos, de início da primeira e da segunda clivagem e de intervalo entre esses eventos, além da observação dos corpúsculos polares. Resultados: Dados mostraram que 84,6% dos embriões apresentaram um padrão de ciclo celular assincrônico e a inviabilidade comprovada com os resultados do PGD, 7,7% dos embriões considerados normais nos resultados do PGD mostraram ter ciclos celulares fora dos padrões e 7,7% dos embriões com ciclo celular normal apresentaram ao PGD anomalias múltiplas. As diferenças foram estatisticamente significantes para p < 0,05. Embriões com PGD normal podem apresentar um ciclo celular assincrônico, o que afeta sua implantação. Conclusões: O estudo preliminar mostra que os dados obtidos com a metodologia time-lapse, em primeiro momento, podem ser usados para avaliar a qualidade embrionária em conjunto com a avaliação morfológica deles, independentemente dos resultados do PGD, pois alterações no padrão de desenvolvimento celular embrionário parecem afetar a implantação, salvo algumas exceções. Porém, como o N amostral ainda é pequeno, necessita-se de um período maior para a certificação dos eventos observados.

Objective: Investigate whether there is correlation between results obtained with PGD and data obtained monitoring “time lapse” during 3 days of embryonic development. Method: This is a retrospective study in which we assessed embryos ICSI cycles from March to December 2012. Upon completion ICSI, embryos were placed in an incubator with vertical monitoring “time lapse” for 3 days, then embryos were subjected to PGD. The results were based on analysis of the time of appearance of two pronuclei, disappearance, the start time of first and second cleavage and intervals between these events, and the observation of polar bodies. Results: Data show that 84.6% of the embryos exhibited a pattern of cell cycle asynchronous and viability confirmed by the results of PGD, and 7.7% of embryos considered normal results of PGD, proved to have cell cycle asynchronous, 7.7% of embryos with normal cell cycle PGD showed multiple anomalies, differences were statistically significant at p < 0.05. PGD embryos with normal can present asynchronous cell cycle, affecting its implantation. Conclusion: The preliminary study shows that data obtained with method “time-lapse”, can be used to evaluate the embryo quality together with morphological evaluation same regardless outcome of PGD, because changes in pattern development of embryonic cell seem to affect development, with some exceptions. However, such as sample size remains small needs to be a longer period for certification of observed events.

A Comparison of Biomechanical Characteristics and Morphologise between Operative and Nonoperative Treatments fo Tenotomized Rabbit Achilles Tendon over Lapse of Time / 대한정형외과학회잡지

The treatment of Achilles tendon rupture is controversial between surgical repair and conservative cast treatment. This study was attempted to compare the biomechanical and morphological results between operative and nonoperative treatments of experimentally tenotomized rabbit Achilles tendon with the lapse of time. A total of 72 adult rabbits were used. After tenotomizing the Achilles tendons, the subjects were divided into 3 groups according to the time of the initiation of the treatment and each group was subdivided into 2 further subgroups according to the method of treatment; immediate operation and nonoperation, 1 week delayed operation and nonoperation, and 2 weeks delayed operation and nonoperation subgroups. Ten animals from each subgroup were sacrificed after four weeks of cast immobilization and the Achilles tendons were prepared for the gross, biomechanical and histological examinations. At the same time, two animals from each subgroup were examined by Microfil perfusion for microvascular changes in the healed tendons. On biomechanical examination, maximal loads were decreased with the lapse of time in each treatment method, and there were statistical significances between the immediate and 2 weeks delayed operation subgroups, 1 week and 2 weeks delayed operation subgroups, and immediate and 2 weeks delayed nonoperation subgroups. A maximal loads were higher in the operative treatment of each group but there was no statistical significances between operative and nonoperative treatment of each group. The results of absorption energy and stiffness were similar to those of maximal load. In conclusion, these results suggest that a rupture of the Achilles tendon should be treated as early as possible preferably within 1 week, in order to achieve a high tensile strength irrespective of the treatment method. In terms of rerupture, nonoperative treatment is comparable with surgical treatment if a rupture of Achilles tendon is managed within 2 weeks of injury.