RESUMEN
PURPOSE: The purpose of the study was to explore the effect of blastomere biopsy for preimplantation genetic diagnosis (PGD) on the embryos' dynamics, further cleavage, development, and implantation. METHODS: The study group included 366 embryos from all PGD treatments (September 2012 to June 2014) cultured in the EmbryoScope™ time-lapse monitoring system. The control group included all intracytoplasmic sperm injection (ICSI) embryos cultured in EmbryoScope™ until day 5 during the same time period (385 embryos). Time points of key embryonic events were analyzed with an EmbryoViewer™. RESULTS: Most (88 %) of the embryos were biopsied at ≥8 cells. These results summarize the further dynamic development of the largest cohort of biopsied embryos and demonstrate that blastomere biopsy of cleavage-stage embryos significantly delayed compaction and blastulation compared to the control non-biopsied embryos. This delay in preimplanation developmental events also affected postimplantation development as observed when the dynamics of non-implanted embryos (known implantation data (KID) negative) were compared to those of implanted embryos (KID positive). CONCLUSION: Analysis of morphokinetic parameters enabled us to explore how blastomere biopsy interferes with the dynamic sequence of developmental events. Our results show that biopsy delays the compaction and the blastulation of the embryos, leading to a decrease in implantation.
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Blastómeros/ultraestructura , Implantación del Embrión/genética , Desarrollo Embrionario/genética , Diagnóstico Preimplantación , Biopsia , Fase de Segmentación del Huevo/metabolismo , Técnicas de Cultivo de Embriones , Transferencia de Embrión/métodos , Femenino , Fertilización In Vitro/métodos , Humanos , Embarazo , Inyecciones de Esperma IntracitoplasmáticasRESUMEN
BACKGROUND: Monocyte-platelet aggregates (MPAs) represent the crossroads between thrombosis and inflammation, and targeting this axis may suppress thromboinflammation. While antiplatelet therapy (APT) reduces platelet-platelet aggregation and thrombosis, its effects on MPA and platelet effector properties on monocytes are uncertain. OBJECTIVES: To analyze the effect of platelets on monocyte activation and APT on MPA and platelet-induced monocyte activation. METHODS: Agonist-stimulated whole blood was incubated in the presence of P-selectin, PSGL1, PAR1, P2Y12, GP IIb/IIIa, and COX-1 inhibitors and assessed for platelet and monocyte activity via flow cytometry. RNA-Seq of monocytes incubated with platelets was used to identify platelet-induced monocyte transcripts and was validated by RT-qPCR in monocyte-PR co-incubation ± APT. RESULTS: Consistent with a proinflammatory platelet effector role, MPAs were increased in patients with COVID-19. RNA-Seq revealed a thromboinflammatory monocyte transcriptome upon incubation with platelets. Monocytes aggregated to platelets expressed higher CD40 and tissue factor than monocytes without platelets (p < 0.05 for each). Inhibition with P-selectin (85% reduction) and PSGL1 (87% reduction) led to a robust decrease in MPA. P2Y12 and PAR1 inhibition lowered MPA formation (30 and 21% reduction, p < 0.05, respectively) and decreased monocyte CD40 and TF expression, while GP IIb/IIIa and COX1 inhibition had no effect. Pretreatment of platelets with P2Y12 inhibitors reduced the expression of platelet-mediated monocyte transcription of proinflammatory SOCS3 and OSM. CONCLUSIONS: Platelets skew monocytes toward a proinflammatory phenotype. Among traditional APTs, P2Y12 inhibition attenuates platelet-induced monocyte activation.
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COVID-19 , Trombosis , Humanos , Plaquetas/metabolismo , Inflamación/metabolismo , Monocitos/metabolismo , Selectina-P/metabolismo , Activación Plaquetaria , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Glicoproteína IIb de Membrana Plaquetaria/metabolismo , Receptor PAR-1/metabolismo , Trombosis/metabolismoRESUMEN
Patients with HIV exhibit platelet activation and increased risk of cardiovascular disease, the prevention of which is not fully known. Fifty-five HIV-positive patients were randomized to clopidogrel, aspirin, or no-treatment for 14 days, and the platelet phenotype and ability to induce endothelial inflammation assessed. Clopidogrel as opposed to aspirin and no-treatment reduced platelet activation (P-selectin and PAC-1 expression). Compared with baseline, platelet-induced proinflammatory transcript expression of cultured endothelial cells were reduced in those assigned to clopidogrel, with no change in the aspirin and no-treatment arms. In HIV, clinical trials of clopidogrel to prevent cardiovascular disease are warranted. (Antiplatelet Therapy in HIV; NCT02559414).
RESUMEN
Spermatogenesis is regulated by a complex network of posttranslation modifications. Sumoylation (a modification by small ubiquitin-like modifiers, or SUMO proteins) was identified as an important cellular event in different cell types. SUMO proteins are highly expressed in the testis, and their role during spermatogenesis has begun to be elucidated. Given the important role of sumoylation in the regulation of mitosis and cancer progression in other tissues, the aim of the current study was to identify the targets of SUMO in proliferating mouse spermatogonia and human seminoma tissues and to initially examine the level of sumoylation in relation to the proliferative activity of the tissues. Using freshly purified spermatogonia and C18-4 spermatogonia cell line, mass spectrometry analysis identified several SUMO targets implicated into the proliferation of spermatogonia (such as heat shock protein 60 [HSP60] and prohibitin). Tissue array and western blot approaches showed that SUMO expression is a prominent feature of human seminomas and that the proliferative activity of the tumor tissues was positively correlated with the level of SUMO expression. Downregulation of sumoylation with si-RNA was not sufficient to significantly affect the proliferation of C18-4 spermatogonia; however, SUMO overexpression increased the proliferation rate of the cells. These data suggest that cells are more sensitive to an elevated level of SUMO, and that this situation may lead to an upregulated cellular proliferation and, possibly, cancer. Mass spectrometry analysis identified around a hundred SUMO targets in seminoma samples. Notably, many of the identified proteins (such as proliferating cell nuclear antigen [PCNA], DNA topoisomerase 2-alpha [Top2A], prohibitin, 14-3-3 protein, and others) were implicated in oncogenic transformation and cancer progression.
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Seminoma/metabolismo , Espermatogonias/crecimiento & desarrollo , Sumoilación , Neoplasias Testiculares/metabolismo , Adulto , Anciano , Animales , Western Blotting , Línea Celular , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Seminoma/patología , Espermatogénesis , Espermatogonias/metabolismo , Neoplasias Testiculares/patologíaRESUMEN
Unexplained infertility (*UI) was a common problem before the IVF era. A couple was declared as UI only after they passed all the routine common tests and no reason was found for the infertility. The introduction of ICSI in the IVF clinics enabled the investigation of the quality of the couple's gametes. Thus, the definition of the term UI was refined and the number of couples encompassed by this term decreased dramatically. Zona Pellucida (ZP) incompetence was described as one of the causes of UI. We recently (2004) published a case study of an UI couple due to this cause. The oocytes of the woman collapsed during the preparation of the oocytes for sperm injection. These oocytes suffered from irregular ZP and abnormal appearance. Only after gentle treatment of the oocyte-cumulus complex and ICSI fertilization, embryo development and delivery of normal newborn was achieved. This woman and others who suffer from UI did not conceive in a natural way since the ZP of the ovulated eggs did not bind sperm cells and, thereby, were not fertilized. It only recently became possible to find the reason for the infertility of the couple and the solution, through the use of highly advanced technology in the IVF-ICSI process. One of the reasons for the deformatted ZP is the malfunction of the gene that encodes the 4 glycoproteins, which compose the ZP. This is now under investigation in our Institute.
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Genitales Femeninos/fisiopatología , Infertilidad Femenina/etiología , Zona Pelúcida , Femenino , Fertilización In Vitro , Humanos , Infertilidad Femenina/fisiopatología , Embarazo , Técnicas Reproductivas Asistidas , Inyecciones de Esperma Intracitoplasmáticas , Interacciones Espermatozoide-ÓvuloRESUMEN
Spinal muscular atrophy (SMA) is an autosomal recessive disorder with a carrier frequency of approximately 1 in 40. Approximately 95% of patients have homozygous deletions of exon 7 and/or 8 of the SMN1 gene. Carrier testing for SMA is relatively complex and requires quantitative polymerase chain reaction (PCR) of genomic DNA to determine SMN1 copy number. The purpose of this study was to assess the feasibility of carrier testing for SMA in males, by nested PCR analysis of SMN1 deletions in single sperm cells. A nested PCR method was developed to amplify SMN1 exon 7 in single cells. Restriction enzyme digestion with DraI was used to differentiate between the highly homologous SMN1 and SMN2 genes. Single sperm cells from five known SMA carriers and six noncarriers were analyzed. Among the five carriers, a total of 132 single sperm cells were analyzed and SMN1 exon 7 deletion was detected in 68 cells (51.5%). In contrast, among the six noncarriers, a total of 136 single sperm cells were analyzed. Of these, an apparent SMN1 exon 7 deletion was detected in four sperm cells. This was interpreted as an allele dropout (ADO) rate of 2.9%. We conclude that nested PCR of SMN1 exon 7 is an accurate and reproducible method for detection of SMA male carriers with a SMN1 deletion.
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Secuencia de Bases , Trastornos de los Cromosomas/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Heterocigoto , Atrofia Muscular Espinal/genética , Proteínas del Tejido Nervioso/genética , Proteínas de Unión al ARN/genética , Eliminación de Secuencia/genética , Espermatozoides , Análisis Mutacional de ADN/métodos , Exones/genética , Femenino , Genes Recesivos/genética , Humanos , Masculino , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Proteínas del Complejo SMN , Sensibilidad y Especificidad , Espermatozoides/citología , Proteína 1 para la Supervivencia de la Neurona Motora , Proteína 2 para la Supervivencia de la Neurona MotoraRESUMEN
OBJECTIVE: To analyze whether the cleavage pattern redefined for all cleavage stages according to the relative blastomere size as a function of cell number has an additive value in predicting implantation potential of day-2 and day-3 embryos. DESIGN: Retrospective analysis of standard embryo morphologic parameters (cleavage rate and degree of fragmentation) supplemented by cleavage pattern findings of 347 implanted embryos compared with those of a matched control group of 307 embryos that failed to implant. SETTING: University-based tertiary medical center. PATIENT(S): Two hundred and nine women with successful implantation and 181 controls matched for age and demographic parameters with failed implantation. INTERVENTION(S): In vitro fertilization (IVF), embryo assessment, and embryo transfer. MAIN OUTCOME MEASURE(S): Determination of cleavage patterns in synchronized and unsynchronized cleaving embryos and correlations with implantation outcomes. RESULT(S): Statistically significantly more embryos of the implanted group had good cleavage patterns compared with the failed implantation group (88% vs. 70%). A good cleavage pattern predicted implantation outcome even for nonsynchronized cleaving blastomeres at three, five, six, and seven cells (79% vs. 59%). Regression analysis demonstrated that adding cleavage pattern to the scoring system increased our ability to predict implantation in the training set; the area under the curve was the highest (0.707) as was the proportion of correct classification (>70%) when the cleavage pattern was assessed on both days 2 and 3. CONCLUSION(S): When combined with measurements of the cleavage rate and degree of fragmentation, the cleavage pattern refines our ability to predict the likelihood of implantation, representing a definitive tool in the selection of top-quality embryos.
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Blastómeros/citología , Fase de Segmentación del Huevo , Implantación del Embrión , Fertilización In Vitro , Recuento de Células , Femenino , Humanos , Embarazo , Pronóstico , Análisis de Regresión , Estudios RetrospectivosRESUMEN
OBJECTIVE: To investigate genetic, molecular and functional aspects of human zona pellucida (ZP) in oocytes with an abnormal appearance. STUDY DESIGN: The study included three women with unexplained infertility whose oocytes had an abnormal ZP appearance and the mother and fertile sister of one of them. The coding exons and their flanking intron regions of the four ZP genes and the regulatory element for the ZP3 gene were sequenced. Immunofluorescence staining of discarded oocytes using monoclonal antibodies against recombinant human ZP glycoproteins and a hemizona assay were performed. RESULTS: No new mutations were observed in the ZP1 (12 exons), ZP2 (19 exons), ZP3 (9 exons), ZP4 (12 exons) genes or in the ZP3 regulatory element of the three studied women. Sequencing of the genes revealed eight synonymous and non-synonymous reported polymorphisms only in ZP1, ZP2 and ZP3. Immunofluorescence staining of the discarded oocytes of two women showed clear and strong staining of the ZP1, ZP2 and ZP4 proteins, but weak staining of the ZP3 protein, although their ZP displayed normal sperm binding ability in the hemizona assay. Intracytoplasmic sperm injection yielded good pregnancy outcomes, even though few injected oocytes developed normally up to day 3. CONCLUSIONS: The abnormal oocyte ZP appearance in the three study women may not have been due to the genetic changes in the ZP genes. Moreover, sperm binding was normal despite low ZP3 staining observed, suggesting that ZP3 profile may play a subordinate role in the reported cases. Our findings support previous studies which claim that abnormal oocyte morphology is not associated with a decrease in fertilization rates or birth outcomes in couples undergoing intracytoplasmic sperm injection.
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Proteínas del Huevo/genética , Infertilidad Femenina/terapia , Glicoproteínas de Membrana/genética , Enfermedades del Ovario/genética , Polimorfismo Genético , Receptores de Superficie Celular/genética , Elementos Reguladores de la Transcripción , Zona Pelúcida/patología , Adulto , Análisis Mutacional de ADN , Ectogénesis , Proteínas del Huevo/química , Proteínas del Huevo/metabolismo , Exones , Salud de la Familia , Femenino , Humanos , Infertilidad Femenina/etiología , Nacimiento Vivo , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/metabolismo , Mutación , Oocitos/metabolismo , Oocitos/patología , Enfermedades del Ovario/metabolismo , Enfermedades del Ovario/patología , Enfermedades del Ovario/fisiopatología , Embarazo , Receptores de Superficie Celular/química , Receptores de Superficie Celular/metabolismo , Inyecciones de Esperma Intracitoplasmáticas , Interacciones Espermatozoide-Óvulo , Zona Pelúcida/metabolismo , Glicoproteínas de la Zona PelúcidaRESUMEN
Human embryonic stem cells (HESCs) carrying specific mutations potentially provide a valuable tool for studying genetic disorders in humans. One preferable approach for obtaining these cell lines is by deriving them from affected preimplantation genetically diagnosed embryos. These unique cells are especially important for modeling human genetic disorders for which there are no adequate research models. They can be further used to gain new insights into developmentally regulated events that occur during human embryo development and that are responsible for the manifestation of genetically inherited disorders. They also have great value for the exploration of new therapeutic protocols, including gene-therapy-based treatments and disease-oriented drug screening and discovery. Here, we report the establishment of 15 different mutant human embryonic stem cell lines derived from genetically affected embryos, all donated by couples undergoing preimplantation genetic diagnosis in our in vitro fertilization unit. For further information regarding access to HESC lines from our repository, for research purposes, please email dalitb@tasmc.health.gov.il.
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Células Madre Embrionarias/citología , Células Madre Embrionarias/metabolismo , Enfermedades Genéticas Congénitas/genética , Mutación/genética , Diagnóstico Preimplantación/métodos , Blastocisto/citología , Blastocisto/metabolismo , Diferenciación Celular/genética , Línea Celular , Ensayo de Unidades Formadoras de Colonias , Regulación del Desarrollo de la Expresión Génica , Humanos , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/metabolismo , Reproducibilidad de los ResultadosRESUMEN
We report on the establishment of a human embryonic stem cell (HESC) line from a preimplantation fragile X-affected embryo and demonstrate its value as an appropriate model to study developmentally regulated events that are involved in the pathogenesis of this disorder. Fragile X syndrome results from FMR1 gene inactivation due to a CGG expansion at the 5'UTR region of the gene. Early events in FMR1 silencing have not been fully characterized due to the lack of appropriate animal or cellular models. Here we show that, despite the presence of a full mutation, affected undifferentiated HESCs express FMR1 and are DNA unmethylated. However, epigenetic silencing by DNA methylation and histone modification occurs upon differentiation. Our unique cell system allows the dissection of the sequence by which these epigenetic changes are acquired and illustrates the importance of HESCs in unraveling developmentally regulated mechanisms associated with human genetic disorders.
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Blastocisto/patología , Diferenciación Celular/genética , Células Madre Embrionarias/patología , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/genética , Síndrome del Cromosoma X Frágil/genética , Mutación , Diagnóstico Preimplantación , Animales , Blastocisto/metabolismo , Línea Celular , Metilación de ADN , Células Madre Embrionarias/metabolismo , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/metabolismo , Síndrome del Cromosoma X Frágil/diagnóstico , Síndrome del Cromosoma X Frágil/metabolismo , Síndrome del Cromosoma X Frágil/patología , Silenciador del Gen , Histonas/metabolismo , Humanos , Ratones , Ratones SCID , Teratoma/genética , Teratoma/patología , Células Tumorales CultivadasRESUMEN
OBJECTIVE: Canavan disease is an autosomal recessive disorder which is relatively common in Ashkenazi Jews. It is characterized by developmental delay, severe hypotonia and early death, and is caused by a deficiency of aspartoacylase which is encoded by the ASPA gene. In Ashkenazi Jews, one major mutation (E285A) and one minor mutation (Y231X) account for the majority of cases. The objective of this study was to develop a preimplantation genetic diagnosis (PGD) protocol for Canavan disease. METHODS: Two carrier couples requested PGD for Canavan disease. In 1 couple each was a carrier of a different ASPA mutation (Y231X and E285A). In the other couple both were carriers of the minor mutation (Y231X). A single-cell duplex nested polymerase chain reaction (PCR) protocol was first developed on single leukocytes obtained from the known carrier parents. Following verification in single leukocytes, clinical PGD was offered to both couples. RESULTS: We evaluated 115 single leukocytes from known carriers and found an allele drop out rate of 1.7% for the fragment harboring the Y231X mutation and 0% for the fragment harboring the E285A mutation. One cycle of PGD was performed in each family. In the first, 11 embryos were successfully analyzed and 4 were found to be affected. Two unaffected embryos were transferred, but no pregnancy resulted. In the other family, 4 embryos were analyzed, 1 was affected, 2 were heterozygotes and 1 was homozygous normal. Following transfer of 2 unaffected embryos, a singleton pregnancy resulted, currently ongoing at 18 weeks gestational age. Amniocentesis performed at 16 weeks confirmed the diagnosis. CONCLUSION: Reliable PGD for Canavan disease is possible using a single-cell nested PCR approach.
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Enfermedad de Canavan/diagnóstico , Enfermedad de Canavan/genética , Pruebas Genéticas/métodos , Reacción en Cadena de la Polimerasa , Diagnóstico Preimplantación/métodos , Femenino , Pruebas Genéticas/normas , Humanos , Judíos/genética , Leucocitos , Embarazo , Diagnóstico Preimplantación/normas , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVES: Duchenne muscular dystrophy (DMD) is a lethal X-linked recessive disorder with an incidence of approximately 1 in 3500 males, caused by mutation in the DMD gene. About 2/3 of DMD cases are caused by gross DMD gene deletion mutations. The purpose of this study was to develop a series of single-cell multiplex-nested PCR protocols for preimplantation genetic diagnosis (PGD) of the most prevalent DMD deletions. METHODS: The protocols were developed on single blood leukocytes from normal males and females and patients with known DMD gene deletion. In the first reaction, 2 of 11 different primer sets (exons 4, 8, 12, 13, 17, 46, 47, 49, 50, 52 and intron 52) were used to allow the simultaneous amplification of different DMD loci and the SRY gender marker, in a single triplex-nested polymerase chain reaction (PCR). Aliquots of this reaction were then subjected to nested PCR in which each locus was amplified individually. Following the successful establishment of single-cell triplex-nested PCR in single leukocytes, the technique was employed in five clinical PGD cases. RESULTS: For each DMD locus, more than 50 single leukocytes from healthy controls and more than 100 single leukocytes from affected individuals with known deletions were analyzed. Amplification efficiency for each tested locus was 98-100%. The false-negative rates for each analysis taken separately was <1%. Taken together, however, the results of the triplex-nested PCR analysis had a false-negative rate of 0%. No contamination was detected in all wash-drop blanks tested. We subsequently performed 18 PGD cycles in 5 DMD carriers. A total of 156 embryos were biopsied and successfully analyzed. Of these, 39 affected embryos were detected and 50 unaffected embryos were transferred (mean = 2.9 +/- 1.1 embryos per cycle). These resulted in three biochemical pregnancies and three clinical pregnancies, all of which have culminated in the birth of normal offspring. CONCLUSION: Triplex-nested PCR using 2 of 11 DMD loci and the SRY gender marker allow PGD for >90% of DMD families with known deletions. These protocols are associated with a high amplification efficiency and accuracy.
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Análisis Citogenético/métodos , Eliminación de Gen , Distrofia Muscular de Duchenne/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Diagnóstico Preimplantación , Estudios de Casos y Controles , Transferencia de Embrión , Reacciones Falso Negativas , Femenino , Humanos , Leucocitos , Masculino , Distrofia Muscular de Duchenne/sangre , Distrofia Muscular de Duchenne/genética , Reacción en Cadena de la Polimerasa/normas , Embarazo , Diagnóstico Preimplantación/éticaRESUMEN
OBJECTIVE: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder caused in most patients by homozygous deletion of the SMN1 gene. For a carrier couple at a 25% risk of affected offspring, preimplantation genetic diagnosis (PGD) offers an alternative to prenatal diagnosis and termination of affected pregnancies. Our objective was to develop an accurate and reliable single-cell multiplex nested PCR analysis for PGD of SMA. METHODS: The method was developed on single blood leukocytes, obtained from healthy controls and an adult SMA type III patient with a known homozygous deletion of SMN1 exon 7 and 8. Multiplex nested PCR on single cells was used to co-amplify exons 7 and 8 of SMN. Additional multiplexing was performed with the ZFX/ZFY gene for sexing. Following successful establishment of the multiplex nested PCR protocol in single leukocytes, the technique was employed for PGD in 4 patients for a total of 7 cycles. In 2 patients, sexing was simultaneously performed using ZFX/ZFY. RESULTS: 220 single leukocytes from a normal individual and 220 from an SMA patient were analyzed. Exon 7 of SMN1 was amplified in 99% of normal single leukocytes and in none of the SMA-affected leukocytes. Exon 7 of SMN2 was amplified in 100% of both normal and SMA-affected leukocytes. Exon 8 of SMN1 was amplified in 98% of normal cells and in none of the SMA-affected leukocytes. Exon 8 of SMN2 was amplified in 96% of both normal and SMA-affected leukocytes. Amplification efficiency was 99% for ZFX/ZFY. There were no false-negative results and no contamination was detected in all wash-drop blanks tested. Seven PGD cycles were performed in 4 SMA-carrier couples with successful molecular analysis of 34 embryos and a total of 15 normal embryos transferred in 7 cycles. One clinical pregnancy has resulted in the delivery of a healthy male. Amniocentesis performed at 17 weeks confirmed the correct diagnosis for both SMA and sexing. CONCLUSIONS: These results suggest that our multiplex nested PCR protocol offers an efficient and accurate method for PGD of SMA while enabling the simultaneous analysis of an additional loci.
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Análisis Citogenético/métodos , Atrofia Muscular Espinal/diagnóstico , Atrofia Muscular Espinal/genética , Reacción en Cadena de la Polimerasa/métodos , Diagnóstico Preimplantación/métodos , Adulto , Femenino , Humanos , Masculino , Oocitos/fisiología , EmbarazoRESUMEN
PURPOSE: To compare the efficacy of two commercially available in vitro fertilization (IVF) and embryo culture media systems: the glucose-free P1 Medium supplemented with 20% synthetic serum substitute (SSS) (Irvine Scientific), and the Cook IVF Medium (Cook, Australia). METHODS: A prospective randomized study. Medical center-based IVF Unit affiliated to the Faculty of Medicine of Tel Aviv University. IVF patients were randomly assigned to either P1 Medium supplemented with 20% SSS (182 patients, 196 cycles) or Cook Medium (167 patients, 179 cycles). RESULTS: Fertilization rates were similar with both media (52.3 +/- 26.1 and 53.8 +/- 27.6, respectively). Likewise, no difference was found in morphological characteristics and grading of cultured embryos. However, a significantly higher proportion of the embryos incubated in the P1 Medium reached the four-cell stage on day 2 or the 6-cell stage on day 3 postfertilization, compared to those incubated in Cook Medium (54.3% vs. 41.9%, p < 0.0001). Clinical pregnancy and delivery rates were improved when oocytes and embryos were cultured in P1 Medium. Finally, Implantation rate was significantly higher in the P1 Medium Group (9.9% vs. 6%, respectively). CONCLUSIONS: Our results suggest that the P1 Medium may be associated with a higher embryo cleavage rate and improved implantation rates compared to the Cook IVF Medium.