A developmentally programmed splicing failure contributes to DNA damage response attenuation during mammalian zygotic genome activation.

Transition from maternal to embryonic transcriptional control is crucial for embryogenesis. However, alternative splicing regulation during this process remains understudied. Using transcriptomic data from human, mouse, and cow preimplantation development, we show that the stage of zygotic genome activation (ZGA) exhibits the highest levels of exon skipping diversity reported for any cell or tissue type. Much of this exon skipping is temporary, leads to disruptive noncanonical isoforms, and occurs in genes enriched for DNA damage response in the three species. Two core spliceosomal components, Snrpb and Snrpd2, regulate these patterns. These genes have low maternal expression at ZGA and increase sharply thereafter. Microinjection of Snrpb/d2 messenger RNA into mouse zygotes reduces the levels of exon skipping at ZGA and leads to increased p53-mediated DNA damage response. We propose that mammalian embryos undergo an evolutionarily conserved, developmentally programmed splicing failure at ZGA that contributes to the attenuation of cellular responses to DNA damage.

Dependence of the Magnetization Process on the Thickness of Fe70Pd30 Nanostructured Thin Film.

Fe-Pd magnetic shape-memory alloys are of major importance for microsystem applications due to their magnetically driven large reversible strains under moderate stresses. In this context, we focus on the synthesis of nanostructured Fe70Pd30 shape-memory alloy antidot array thin films with different layer thicknesses in the range from 20 nm to 80 nm, deposited onto nanostructured alumina membranes. A significant change in the magnetization process of nanostructured samples was detected by varying the layer thickness. The in-plane coercivity for the antidot array samples increased with decreasing layer thickness, whereas for non-patterned films the coercive field decreased. Anomalous coercivity dependence with temperature was detected for thinner antidot array samples, observing a critical temperature at which the in-plane coercivity behavior changed. A significant reduction in the Curie temperature for antidot samples with thinner layer thicknesses was observed. We attribute these effects to complex magnetization reversal processes and the three-dimensional magnetization profile induced by the nanoholes. These findings could be of major interest in the development of novel magnetic sensors and thermo-magnetic recording patterned media based on template-assisted deposition techniques.

Administration of the nonsteroidal anti-inflammatory drug tolfenamic acid at embryo transfer improves maintenance of pregnancy and embryo survival in recipient mice.

PURPOSE: To evaluate the effect of the nonsteroidal anti-inflammatory drugs tolfenamic acid and flunixin meglumine in pregnancy rate and embryo survival of recipient mice subjected to embryo transfer. METHODS: A total of 142 recipient females were transferred with 2,931 embryos and treated with a single injection of tolfenamic acid (1 mg/kg; n = 54 females with 1,129 embryos), flunixin meglumine (2.5 mg/kg; n = 46 females with 942 embryos), or bi-distilled water (10 mL/kg) as control group (n = 42 females with 860 embryos). Pregnancy was checked 2 weeks after embryo transfer, delivery was registered on the due date, and litter size was recorded on Day 7 after birth. RESULTS: Pregnancy rate of tolfenamic acid treated females was significantly higher than flunixin group (P < 0.05) and showed a tendency to be higher when compared to the control group (P = 0.06). The number of pups born from transferred embryos in pregnant females was significantly higher for both treatment groups compared to controls (P < 0.05). Number of pups from total transferred embryos was higher for both treatment groups (P < 0.05) when compared to controls. CONCLUSION: The use of tolfenamic acid at the time of embryo transfer improves both pregnancy rate and number of live pups in recipient mice, with optimal effects observed with flunixin meglumine. We suggest that the use of tolfenamic acid has beneficial effects on the maintenance of pregnancy and embryo survival in recipient mice, which should be taken into account for further studies in other mammalian females.

Genotipo de los genes VKORC1 y CYP2C9 en la respuesta individual a la warfarina / Genotype of the VKORC1 and CYP2C9 genes in individual response to warfarin

Introducción: la warfarina es un fármaco ampliamente utilizado como anticoagulante oral. Su estrecho rango terapéutico y marcada variabilidad interindividual en la respuesta requieren un control riguroso en su administración para evitar accidentes hemorrágicos. Objetivos: correlacionar las variantes genéticas de CYP2C9*2 y *3 y VKORC1 (C1173T) con la respuesta y los efectos adversos. Material y método: los genotipos CYP2C9*1, *2, *3, y VKORC1 fueron obtenidos por PCR-RFLP y los resultados analizados usando el paquete estadístico SPSS 12.0. Resultados: hay una tendencia a la reducción de dosis en relación con la presencia de alelos polimórficos. Los portadores de CYP2C9*3 requirieron la menor dosis de mantenimiento, seguidos por los portadores de CYP2C9*2 y homocigotos CYP2C9 *1, en ese orden, (4,4±1,0 vesus 5,4±2,3 versus 7,0±3,6 mg/d, p=0,03). Los portadores CYP2C9*3 tuvieron, además, un aumento del riesgo de sobreanticoagulación y requirieron casi el doble de ajustes de dosis para lograr una adecuada anticoagulación. Para VKORC1, los homocigotas T/T necesitaron la dosis más baja, seguidos por los heterocigotas C/T y homocigotas C/C, en ese orden (3,6±0,6 versus 5,5±0,5 y 7,9±0,7 mg/d, p <0,001). Los pacientes T/T tuvieron un mayor riesgo de sobreanticoagulación que los C/T y C/C. El genotipo T/T de VKORC1 produce en todas las combinaciones con CYP2C9 una disminución cercana a 50% de la dosis diaria de warfarina. Conclusiones: se confirma una sensibilidad aumentada a la warfarina en pacientes portadores de alelos *2 y *3 de CYP2C9, y T de VKORC1. Se demuestra un efecto combinado (aproximadamente aditivo) de los alelos variantes de ambos genes.

Summary Introduction: warfarin is a widely used oral anticoagulant. Its narrow therapeutic range (NTR) and its large interindividual variability requires strict control when administered to avoid hemorrhagic accidents. Objectives: to correlate CYP2C9*2 and *3 and VKORC1 (C1173T) genetic variants with response and adverse side effects. Method: CYP2C9*1, *2, *3, and VKORC1 genotypes were obtained by a commonly used PCR-RFLP procedure. The results were analyzed using SPSS 12.0. statistical package. Results: there is a tendency to reduce the dosage in connection with the presence of polymorphic alleles. CYP2C9*3 carriers require the lower maintenance dosage, followed by CYP2C9*2 carriers and then by CYP2C9 *1 homozygotes (4.4±1.0 versus 5.4±2.3 versus 7.0±3.6 mg/d, p=0.03). CYP2C9*3 carriers also showed an increase in the anticoagulation risk, which required almost twice the number of dose adjustments to achieve appropriate anticoagulation. As to VKORC1, T/T homozygotes needed the lowest dose, followed by the C/T heterozygotes, and then by the C/C homozygotes (3.6±0.6 versus 5.5±0.5 and 7.9±0.7 mg/d, p <0,001). Risk of overcoagulation was higher in T/T patients than in C/T or C/C patients. T/T genotype of VKORC1 causes a decrease of nearly 50% in the warfarine daily dosage for all combinations with CYP2C9. Conclusions: we confirmed an increased sensitivity to warfarine in patient carriers of *2 and *3 CYP2C9 alleles and T VKORC1 alleles. We showed a combined effect (approximately accumulative) of variant alleles in both genes.

Résumé Introduction: la warfarine est une drogue très utilisée comme anticoagulant oral. Étant donné son restreint rang thérapeutique et sa remarquable variabilité interindividuelle, un contrôle rigoureux du dosage s’avère indispensable afin d’éviter des accidents hémorragiques. Objectifs: mettre en rapport les variantes génétiques de CYP2C9*2 et *3 et VKORC1 (C1173T) avec la réponse et les effets adverses. Matériel et méthode: les génotypes CYP2C9*1,*2,*3, et VKORC1 résultent par PCR-RFLP et les résultats analysés au moyen du paquet statistique SPSS 12.0. Résultats: il existe une tendance à réduire la dose par la présence d’allèles polymorphiques. Les porteurs de CYP2C9*3 ont requis une dose plus basse de maintien, puis les porteurs de CYP2C9*2 et homozygotes CYP2C9*1, dans cet ordre, (4,4±1,0 versus 7,0±3,6 mg/d, p=0,03). Les porteurs CYP2C9*3 ont aussi subi une augmentation du risque de suranticoagulation et ont requis presque le double d’ajustement de la dose pour atteindre une anticoagulation adéquate. Pour VKORC1, les homozygotes T/T ont requis une dose plus basse, suivis des hétérozygotes C/T et des homozygotes C/C, dans cet ordre (3,6±0,6 versus 5,5±0,5 et 7,9±0,7 mg/d, p <0,001). Les patients T/T ont subi un plus grand risque de suranticoagulation que les C/T et C/C. Le génotype T/T de VKORC1 provoque dans toutes les combinaisons avec CYP2C9 une diminution de 50% environ de la dose par jour de warfarine. Conclusions: on confirme une sensibilité augmentée à la warfarine chez les patients porteurs d’allèles *2 et *3 de CYP2C9, et T de VKORC1. Un effet combiné (à peu près aditif) des allèles variantes des deux gènes reste évident.

Resumo Introdução: a warfarina é uma droga muito usada como anticoagulante oral. Sua administração deve ser controlada rigorosamente para evitar acidentes hemorrágicos devido a uma aplicação terapêutica restrita e a variação da resposta individual. Objetivos: correlacionar as variantes genéticas de CYP2C9*2 e *3 y VKORC1 (C1173T) com a resposta e os efeitos adversos. Material e método: os genótipos CYP2C9*1, *2, *3, e VKORC1 foram obtidos por PCR-RFLP e os resultados foram analisados usando o programa SPSS 12.0. Resultados: observa-se uma tendência à redução da dose na presença de alelos polimórficos. Os portadores de CYP2C9*3 necessitam doses de manutenção mais baixas, seguidos pelos portadores de CYP2C9*2 e homozigotos CYP2C9 *1 (4,4±1,0 versus 5,4±2,3 versus 7,0±3,6 mg/d, p=0,03). Os portadores de CYP2C9*3 apresentaram também um aumento do risco de sobreanticoagulaçao e necessitam o dobro do número de ajustes de dose para obter uma anticoagulaçao adequada. Para VKORC1, os homozigotos T/T necessitaram a dose mais baixa, seguidos pelos heterozigotos C/T e homozigotos C/C (3,6±0,6 versus 5,5±0,5 y 7,9±0,7 mg/d, p <0,001). Os pacientes T/T apresentaram um risco maior de sobreanticoagulação que os C/T e C/C. O genótipo T/T de VKORC1 produz uma redução de aproximadamente 50% da dose diária de warfarina em todas as combinações com CYP2C9. Conclusões: confirma-se a sensibilidade aumentada à warfarina de pacientes portadores de alelos *2 y *3 de CYP2C9, e T de VKORC1. Demonstram-se um efeito combinado (aproximadamente aditivo) dos alelos variantes de ambos os genes.