First pilot study of maternal spindle transfer for the treatment of repeated in vitro fertilization failures in couples with idiopathic infertility.

OBJECTIVES: To gain insights into the technical feasibility of maternal spindle transfer (MST) applied in the context of repeated in vitro fertilization (IVF) failures for the treatment of idiopathic infertility. DESIGN: A prospective pilot study. SETTING: IVF center. PATIENT(S): Twenty-five infertile couples with multiple previous unsuccessful IVF cycles (range, 3-11), no previous pregnancy, and no history of mitochondrial DNA (mtDNA) disease participated. The study focused on women <40 years, with previous IVF attempts characterized by a pattern of low fertilization rates and/or impaired embryo development. Couples with severe male-factor infertility were not eligible. Oocyte donors with previous successful IVF outcomes were matched with patients according to standard practice. INTERVENTION(S): We performed MST by transferring metaphase II spindles from the patients' oocytes into the previously enucleated donor oocytes, followed by intracytoplasmic sperm injection, in vitro embryo culture, blastocyst biopsy, and vitrification. Only euploid blastocysts were considered for embryo transfer. MAIN OUTCOME MEASURE(S): Outcome measures included oocyte fertilization, blastocyst development, clinical pregnancy and live birth, incidence of mitochondrial carryover and potential mtDNA reversal, as well as general health of the children born. RESULT(S): Twenty-eight MST cycles produced 6 children (19 embryo transfers, 7 clinical pregnancies). Pediatric follow-up of the children, performed at intervals from birth to 12-24 months of age, revealed their development to be unremarkable. DNA fingerprinting confirmed that the nuclear DNA of MST children was inherited from both parents, without any contribution from the oocyte donor. For 5 of the children, mtDNA was derived almost exclusively (>99%) from the donor. However, 1 child, who had similarly low mtDNA carryover (0.8%) at the blastocyst stage, showed an increase in the maternal mtDNA haplotype, accounting for 30% to 60% of the total at birth. CONCLUSION(S): This pilot study provides the first insights into the feasibility of applying MST for patients with idiopathic infertility and repeated IVF failures. Reconstructed oocytes produced embryos capable of implanting, developing to term and producing apparently healthy newborns/children. However, claims concerning the efficacy of MST with respect to infertility treatment would be premature considering the limitations of this study. Importantly, mtDNA reversal was detected in one child born after MST, a finding with possible implications for mitochondrial replacement therapies. CLINICAL TRIAL REGISTRATION NUMBER: Pilot trial registry number, ISRCTN11455145. The date of registration: 20/02/2018. The date of enrolment of the first patients: 18/03/2018.

Parameters of the Mouse Embryo Assay that affect detection of peroxides in mineral oil.

RESEARCH QUESTION: Can culture conditions influence the sensitivity of a Mouse Embryo Assay and its potential to detect peroxide-related toxicity in mineral oil samples? DESIGN: Protein type and concentration, embryo density and culture dish design were selected as the variables in the culture system with the potential to influence the assay's sensitivity. Fresh 1-cell mouse embryos were cultured under mineral oil samples with known peroxide concentrations. Protein type (human serum albumin [HSA] + α/ß-Globulins versus HSA versus bovine serum albumin [BSA]), concentration (5 mg/ml versus 0.5 mg/ml), embryo density (25 versus 3 µl/embryo) and culture dish (Petri versus micro-well dish) were adjusted to define the culture conditions with the highest sensitivity. RESULTS: High concentrations of peroxides can be easily detected by current quality control standards. However, for oil samples with a lower concentration of peroxides, supplementing the culture medium with 5 mg/ml of HSA + alpha/beta-globulins or with HSA resulted in an increased detection of embryo toxicity compared with when BSA was used as the protein supplement. The sensitivity of the assay was greatly reduced when embryos were cultured in groups and when certain micro-well dishes were used. CONCLUSIONS: Current quality control protocols may not be sensitive enough to identify low concentrations of peroxides, which, if undetected, can increase over time and become potentially harmful during gamete and embryo culture. The different parameters established in this study allow the sensitivity of the Mouse Embryo Assays to be optimized to specifically detect peroxides in mineral oil samples prior to their release into the market and their broad use in human IVF.

Principios básicos para una ruta de formación en la cualificación de los docentes en el diseño y aplicación de recursos educativos digitales / Basic principles for a training pathway in the teaching qualification in terms of design and application of digital educational resources

Resumen La implementación de rutas de formación para los docentes orientados a la cualificación, diseño y aplicación de recursos educativos digitales son necesarios y prioritarios para que las instituciones avancen hacia la transformación cultural requerida en los procesos de enseñanza-aprendizaje con soporte en las TIC. Esto implica una revisión de la construcción de los conceptos de comunidades de aprendizaje, práctica, práctica virtual, zona de desarrollo próximo en la práctica pedagógica, las relaciones en los ambientes virtuales de aprendizaje y las competencias docentes en el uso de las TIC.

Abstract The implementation of teacher training pathways aiming at the qualifica tion, design, and application of digital educational resources are necessary and priority so that institutions move towards the required cultural transformation in the teaching and learning processes with ICT support. This involves a review of the construction of the concept of communities of learning, communities of practice, virtual practice, zone of proximal development in pedagogical practice, relationships in virtual learning environments, and educational expertise in the use of ICT.

Prevalence and resistance pattern of genotype G and H in chronic hepatitis B and HIV co-infected patients in Mexico.

OBJECTIVE: We estimated the prevalence and identified the resistance pattern of HBV genotypes H and G in HBV monoinfected and HIV co-infected patients. MATERIAL AND METHODS: A cross-sectional prevalence and analytic study were performed in chronic hepatitis B patients at the Hospital de Infectología, La Raza National Medical Center in Mexico City. Chronic HBV monoinfected and HIV co-infected patients were included. HBeAg, HBV viral load and genetic analysis of mutations were collected; CD4+ cells count from HIV co-infected patients and HIV RNA were measured. We calculated the prevalence and exact 95% binomial confidence interval and the Odds ratios (OR) with 95% confidence intervals to assess the relationship between the presence of risk factors and HBV genotypes H or G. RESULTS: We enrolled 77 patients, 67 men and 10 women with 37 HIV co-infected patients. The distribution of HBV genotypes was: HBV genotype H 55 (71% [95% CI 60% to 80%]), HBV genotype G 16 (20.7%), HBV genotype F 4 (5.1%) and HBV genotype A 2 (2.6%). The most frequent mutations presented in 8 HIV co-infected patients and one mono-infected patient with antiretroviral therapy (ART) experience were rtM204V and six of them showed genotype G (6/9). Mono-infected HBV patients exposed more probability to HBV genotype H than co-infected HIV patients OR 13.0 (CI 95% 3.40-49.79), p = 0.0001. In contrast co-infected patients presented less possibility to have genotype H, 0.56 (CI 95% 0.42-0.75). CONCLUSIONS: This study confirms the high prevalence of HBV genotype H in Mexico; furthermore, our results suggest that HBV genotype G predominates in co-infected patients. As well, rtM204V and rtL180M mutations are common in HBV-HIV co-infected patients with genotype G and ART experience.

Malaria vector species in Colombia: a review

Here we present a comprehensive review of the literature on the vectorial importance of the major Anopheles malaria vectors in Colombia. We provide basic information on the geographical distribution, altitudinal range, immature habitats, adult behaviour, feeding preferences and anthropophily, endophily and infectivity rates. We additionally review information on the life cycle, longevity and population fluctuation of Colombian Anopheles species. Emphasis was placed on the primary vectors that have been epidemiologically incriminated in malaria transmission: Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. The role of a selection of local, regional or secondary vectors (e.g., Anopheles pseudopunctipennis and Anopheles neivai) is also discussed. We highlight the importance of combining biological, morphological and molecular data for the correct taxonomical determination of a given species, particularly for members of the species complexes. We likewise emphasise the importance of studying the bionomics of primary and secondary vectors along with an examination of the local conditions affecting the transmission of malaria. The presence and spread of the major vectors and the emergence of secondary species capable of transmitting human Plasmodia are of great interest. When selecting control measures, the anopheline diversity in the region must be considered. Variation in macroclimate conditions over a species' geographical range must be well understood and targeted to plan effective control measures based on the population dynamics of the local Anopheles species.

Could the addition of hp-hMG and GnRH antagonists modulate the response in IVF-ICSI cycles?

OBJECTIVE: To assess if the luteinizing hormone/human chorionic gonadotropin present in some gonadotropin formulations may be of benefit in protocols with GnRH antagonists. METHODS: Open, quasi-experimental, multicenter, prospective, parallel-controlled study compared 136 women undergoing in vitro fertilization--intracytoplasmic sperm injection after stimulation with highly purified human menopausal gonadotropin (hp-hMG) (n = 44), recombinant-follicle stimulating hormone (r-FSH) (n = 46), or a combination of both (r FSH + hp-hMG) (n = 46) following an antagonist protocol. Blood determinations were made on day 6 of stimulation and on the day of ovulation induction, with centralized analysis. RESULTS: No differences were found in the ongoing pregnancy rates between groups [37.0% versus 29.5% (hp-hMG) and 23.9% (r-FSH); p = 0.688]. However, the ratio top-quality embryos/retrieved oocytes (TQE/RO) was higher in the combined therapy group (19.6%)--reaching significance versus the r-FSH group (6.5%) (p = 0.008), but not versus hp-hMG (12.3%) (p = 0.137). CONCLUSIONS: An improved TQE/RO ratio was obtained together with a greater percentage of frozen embryos in the patients that incorporated hp-hMG to their stimulation protocol. Despite good results of adding hp-hMG, non statistical differences were found in terms of ongoing pregnancy rate.

Antimitotic treatments for chemically assisted oocyte enucleation in nuclear transfer procedures.

Chemically assisted enucleation has been successfully applied to porcine and bovine oocytes to prepare recipient cytoplasts for nuclear transfer procedures. In this study, the antimitotic drugs demecolcine, nocodazole, and vinblastine were first assessed for their ability to induce the formation of cortical membrane protrusions in mouse, goat, and human oocytes. While only 2% of the treated human oocytes were able to form a protrusion, high rates of protrusion formation were obtained both in mouse (84%) and goat oocytes (92%), once the treatment was optimized for each species. None of the antimitotics applied was superior to the others in terms of protrusion formation, but mouse oocytes treated with vinblastine were unable to restore normal spindle morphology after drug removal and their in vitro development after parthenogenetic activation was severely compromised, rendering this antimitotic useless for chemically assisted enucleation approaches. Aspiration of the protrusions in mouse oocytes treated with demecolcine or nocodazole yielded 90% of successfully enucleated oocytes and allowed the extraction of a smaller amount of cytoplasm than with mechanical enucleation, but both enucleation methods resulted in the depletion of spindle-associated gamma-tubulin from the prepared cytoplasts. Treatment of mouse oocytes with demecolcine or nocodazole had no effect on their in vitro development after parthenogenetic activation, or on their ability to repolymerize a new spindle after the removal of the drug or the reconstruction of the treated cytoplasts with a somatic nucleus. Therefore, demecolcine- and nocodazole-assisted enucleation appears as an efficient alternative to mechanical enucleation, which can simplify nuclear transfer procedures.

Evaluación del triflumuron y la mezcla de Bacillus thuringiensis más B. sphaericus para control de formas inmaduras de Aedes aegypti y Culex quinquefasciatus en sumideros en Cali, Colombia / Evaluation of the triflumuron and the mixture of Bacillus thuringiensis plus B Bacillus sphaericus for control of the immature stages of Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) in catch basins

Introducción. En Cali los sumideros son uno de los principales criaderos de Aedes aegypti y Culex quinquefasciatus que son controlados por la Secretaría de Salud Municipal utilizando el insecticida triflumurón desde 1999. Se sospecha falla al tratamiento. Objetivos. Evaluar la eficacia del Starycide® (Triflumuron) y VectoMax® (mezcla bacteriana de Bacillus thuringiensis var. israelensis + B. sphaericus) en el control de Ae. aegypti y Cx. quinquefasciatus en los sumideros y determinar el efecto residual de una única aplicación deVectoMax®, en épocas de alta y baja pluviosidad.Materiales y métodos. La eficacia de los productos fue medida en 60 sumideros de una zona residencial de Cali por un período de 90 días. La media de individuos inmaduros (larvas y pupas de Ae. aegypti y Cx. quinquefasciatus) fueron obtenidas quincenalmente de 40sumideros intervenidos (20 con Triflumuron y 20 con VectoMax®) y 20 sin tratamiento (grupo testigo). El efecto residual de la mezcla bacteriana se evaluó quincenalmente en 10 sumideros en cada temporada climática evaluada. Resultados. Los sumideros tratados con VectoMax® presentaron diferencias en el promediode estadios inmaduros en ambas especies frente al testigo (p<0,01). En contraste, el tratamiento con triflumuron sólo presentó diferencias en los estadios inmaduros de Ae. aegypti con respecto al testigo (p<0,001). El efecto residual del VectoMax® fue mayor en la época de baja pluviosidad con respecto al testigo (p<0,001). Conclusión. La mezcla bacteriana fue el tratamiento más eficaz en el control de ambasespecies durante el período evaluado (15 días).