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Evaluation of an internal control in a multiplex-PCR assay for sex determination of <i>in vitro</i>-produced bovine embryos

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Advances in Bioscience and Biotechnology 2011年第6期2卷 456-462页

作者： Fabiana de Almeida Rufino Marcelo Marcondes Seneda Alice Fernandes Alfieri Katia cristina Silva-Santos karina cristina puggesi rubin Amauri Alcindo Alfieri 不详

The use of an internal control in a multiplex-PCR assay for sex determination of In Vitro-produced bovine embryos was evaluated in biopsies of groups of 54 fresh and 44 frozen embryos. The internal controls used were the primers BOV 1 and BOV 2, which amplify a product with 626 base pairs (bp) of bovine mitochondrial DNA ND5 gene. The primers BRY.4aF and BRY.4aR were used for bovine Y chromosome sequence amplification. The specificity of multiplex- PCR reactions realized in biopsies corresponding to about 20% of each fresh embryo (10 male and 10 female) by means of confirming the sexing in the remaining embryo content (~80%) presented 100% specificity. Amplicons of the internal control and Y chromosome were both amplified until dilution corresponding to 6.25% of total extracted DNA from a male embryo. Sex determination was possible in 53 (98.1%) fresh embryos and 40 (90.9%) frozen embryos. The products related to the Y chromosome and mitochondrial DNA were simultaneously amplified in 34 (63%) fresh embryos and 27 (61.4%) frozen embryos, showing a male embryo. The female sex, distinguished by internal control amplification only, was detected in 19 (35.2%) and 13 (29.5%) biopsies, respectively, of fresh and frozen embryos. In one (1.8%) and four (9.1%) biopsies of fresh and frozen embryos, respectively, neither product was amplified, most likely due to the absence of embryonic cells or the presence of embryonic cells going through apoptosis. The multiplex-PCR assay developed in this work showed avoided the limitation of a lack of an internal standard, and was also sensitive, specific, and efficient in reaction failure identification. This technique shows great potential for use on a commercial scale in routine sex determination of In Vitro-produced embryos.

关键词： Bovine Embryos In Vitro Production Sexing Multiplex-PCR

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Sexual behavior and male volatile compounds in wild and mass-reared strains of the Mexican fruit fly Anastrepha ludens （Diptera： Tephritidae） held under different colony management regimes

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Insect Science 2016年第1期23卷 105-116页

作者： Carlos Felipe Bosa Leopoldo Cruz-Lopez cristina Silvia Zepeda-Cisneros Javier Valle-Mora karina Guillen-Navarro Pablo Liedo El Colegio de la Frontera Sur Carretera Antiguo Aeropuerto km 2.5 Chiapas 30700 Mexico Campana Nacional Moscas de laFruta Programa Moscafrut SAGARPA-IICA Metapa de Dominguez Chiapas Mexico

We compared the calling and mating behavior and volatile release of wild males Anastrepha ludens （Loew） with males from 4 mass-reared strains：（i） a stan- dard mass-reared colony （control）, （ii） a genetic sexing strain （Tap-7）, （iii） a colony started from males selected on their survival and mating competitiveness abilities （selected）, and （iv） a hybrid colony started by crossing wild males with control females. Selected and wild males were more competitive, achieving more matings under field cage conditions. Mass-reared strains showed higher percentages of pheromone calling males under field conditions except for Tap-7 males, which showed the highest percentages of pheromone calling males under laboratory cage conditions. For mature males of all strains, field-cage calling behavior increased during the last hour before sunset, with almost a 2 fold increase exhibited by wild males during the last half hour. The highest peak mating activity of the 4 mass-reared strains occurred 30 rain earlier than for wild males. By means of solid phase microextraction （SPME） plus gas chromatography-mass spectrometry （GC- MS）, the composition of volatiles released by males was analyzed and quantified, Wild males emitted significantly less amounts of （E,E）-α-farnesene but emitted significantly more amounts of （E,E）-suspensolide as they aged than mass-reared males. Within the 4 mass-reared strains, Tap-7 released significantly more amounts of （E,E）-α-farnesene and hybrid more of （E,E）-suspensolide. Differences in chemical composition could be ex- plained by the intrinsic characteristics of the strains and the colony management regimes. Characterization of calling behavior and age changes of volatile composition between wild and mass-reared strains could explain the differences in mating competitiveness and may be useful for optimizing the sterile insect technique in A. ludens.

关键词： age-dependent changes fruit fly mating pheromone

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Novel Kdr mutations (K964R and A943V) in pyrethroid-resistant populations of Triatoma mazzottii and Triatoma longipennisUoro Mexico and detoxifying enzymes

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Insect Science 2019年第5期26卷 809-820页

作者： Jesus Davila-Barboza O.karina Villanueva-Segura Beatriz Lopez-Monroy Gustavo Ponce-Garcia cristina Bobadilla-Utrera Mayela Montes-Rincon Zinnia J.Molina-Garza Juan I.Arredondo-Jimenez Iram P.Rodriguez-Sanchez Pablo C.Manrique-Saide Adriana E.Flores Universidad Autonoma de Nuevo Leon (UANL) Facultad de Ciencias BiologicasSan Nicolas de los GarzaNLMexico Laboratorio Estatal de Salud Publica de los Servicios de Salud de Veracruz VeracruzMexico Campus de Ciencias Biologicas y Agropecuarias Unidad Colaborativa para Bioensayos EntomologicosUniversidad Autonoma de YucatanMeridaYucatanMexico

Although having five different ways of transmission the vector-borne is the principal way of transmission of Chagas disease, which involves insects of the subfamily Triatominae (Hemiptera: Reduviidae). Nineteen of the 31 species that occur in Mexico are associated with humans, and all are capable of transmitting the disease. Pyrethroids are the insecticides recommended for the control of these vectors in Mexico. We determined the susceptibility to the pyrethroids dcltamethrin and permethrin of peridomestic populations of Triatoma mazzottii Usinger and two populations of Triatoma longipetmis Usinger in comparison with a reference strain for each species. Bioassays were performed for the determination of the LD50 for both field populations and reference strains. A maximum of 27 fold resistance to deltamethrin was observed in T. mazzottii, meanwhile, for permethrin, T. longipennis from Jalisco show the highest value of 3.19 fold. There was significantly increased activity of esterases in field populations in comparison with their corresponding reference strain. The results of the search of kdr mutations related to the resistance to deltamethrin and permethrin in the evaluated species show the presence of mutations in the field populations, as is the case with individuals of T. mazzottii, for which the mutation was found A943V, and for the two populations of T. longipennis included in this study, we report the presence of the kdr mutation K964R. Evaluation of the various mechanisms involved in resistance to pyrethroids in triatomines from Mexico could guide us to the real justification for insecticide resistance monitoring.

关键词： detoxifying enzymes kdr pyrethroids Triatoma longipennis Triatoma mazzottii

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Consequences of Different Hysterectomy Techniques in Sexual Behavior——A Systematic Review

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Open Journal of Obstetrics and Gynecology 2014年第6期4卷 333-341页

作者： Ana Fátima Fernandes Ana Izabel Nicolau Camila Vasconcelos Pricilla Alves Ana karina Pinheiro Namie Sawada cristina Maria Galvao Camila Moreira Department of Nursing Federal University of CearáFortalezaBrazil

We conducted a systematic review of the literature to determine the influence of different hysterectomy techniques in the sexual behavior of women who underwent. We searched CENTRAL (The Cochrane Library), PubMed, SCOPUS, CINAHL and LILACS for studies between the years 1998 and 2010 that prospectively examined the sexuality after hysterectomy. Those that were not randomized controlled clinical trials were excluded. It was not available electronically, it was not related with the outcomes determined, it was not research, but publications before 1998 and study which women realized chemotherapy and radiotherapy. From the 455 identified studies, 9 met inclusion criteria. There was variability in how hysterectomies influence in the sexual behavior. Most of studies considered women who underwent vaginal hysterectomy had fewer changes in sexual behavior. Sexual desire, intercourse frequency and orgasm are the sexual aspects reported by women as more changes experimented after surgery. Findings from this study have implications for healthcare providers and policymakers. Each woman needs an individualized care plan that fits within the context of her life, and there are basic interventions that every woman who underwent hysterectomy should receive. This review draws attention to the need for different care plans for women who underwent different hysterectomy techniques.

关键词： Hysterectomy Sexual Behavior Sistematic Review

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