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Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm

journal of Animal Science and Biotechnology 2017年第2期8卷 370-377页

作者： j.e.hernández-pichardo Y.Ducolomb S.Romo M.e.Kjelland R.Fierro F.Casillas M.Betancourt División de Ciencias Biológicas y de la Salud Departamento de Producción Agrícola y AnimalUniversidad Autónoma Metropolitana-XochimilcoCiudad de MexicoMexico Division de Ciencias Biologicas y de la Salud Departamento de Ciencias de la SaludUniversidad Autónoma Metropolitana-IztapalapaSan Rafael Atlixco 186CP 09340 Ciudad de MexicoMexico Departamento de Ciencias Pecuarias Facultad de EstudiosSuperiores CuautitlanUNAMCiudad de MexicoEstado de MexicoMexico Conservation Genetics&BiotechLLCValley CityNDUSA Division de Ciencias Biologicas y de la Salud Departamento de Biologia de la ReproduccionUniversidad Autonoma Metropolitana-lztapalapaCiudad de MexicoMexico Doctorado en Biologia experimental Universidad Autonoma Metropolitana-lztapalapaCiudad de MexicoMexico

Background： In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up（SU） or swim up ＋ zona pellucida（SU ＋ ZP） binding.Results： experiment 1, 4–20 replicates with total 821 in vitro matured oocytes were chemically activated with ethanol, calcium ionophore or ionomycin, to determine oocyte activation（precense of one PN）. Treatments showed similar results（54, 47, 42 %, respectively） but statistically differents（P 0.05）.Conclusions： Chemical activation induces higher ovine oocyte activation than mechanical activation. ethanol slightly displays higher oocyte activation than calcium ionophore and ionomicine. Sperm selection with SU ＋ ZP increased AR/A and AR/D rates in comparison with SU in fresh and frozen-thawed sperm. According to this, in terms of fertilization rates, chemical activation after ICSI increased oocyte PN formation compared to mechanical activation. Also, fresh sperm treated with SU and SU ＋ ZP were significantly different than frozen-thawed sperm,but between sperm treatments no significant differences were obtained.

关键词： ethanol ICSI Oocyte activation Ovine Pronucleus Zona pellucida

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Implementation of a New Solution for the Preservation of Anatomical Specimens Made of Non-Toxic Substances

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Open journal of Veterinary Medicine 2024年第3期14卷 56-67页

作者： Ma Reyes pichardo-Molinero Samantha jardon-Xicotencatl Misael R. Oliver-González Carlos G. García-Tovar Laboratorio de Apoyo Técnico a la Anatomía Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México Estado de México Mexico Unidad de Investigación Multidisciplinaria L4 (Morfología Veterinaria y Biología Celular) Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México Estado de México Mexico

The preservation of anatomical pieces in Veterinary Anatomy is essential since it is not possible to dissect all domestic species. Most techniques use reagents with high levels of toxicity such as formaldehyde. The objective of this work was to develop a new preservation technique that uses reagents with zero toxicity and that allows obtaining preserved pieces suitable for anatomical studies. The alcohol propylene glycol technique was developed, the method of which uses a fixation step with alcohol, sodium chloride, commercial vinegar and subsequently the impregnation of the preservation solution made from propylene glycol and commercial vinegar, which are non-toxic. As a result of this work, adequately preserved sheep hearts were obtained that preserved their morphology with slight changes in size and weight, which did not affect their external and internal anatomical structure. Its coloration was not substantially affected, remaining a little lighter. The pieces obtained showed flexibility which allowed dissections to be carried out. The time to develop the technique was 20 days. A comparative study was carried out with the phenolated glycerin technique that uses toxic reagents (formaldehyde and phenol) and the pieces obtained with the alcohol propylene glycol technique were of better quality, observing that the pieces with phenolated glycerin tend to darken and are more rigid. And the time to develop the technique is 24 days. In conclusion, a preservation technique for anatomical pieces was developed that allowed the preservation of the organs under study, which allow their use for anatomical studies, and which have been preserved without changes until the time of this publication (8 months) and there are pieces preserved with this technique for 2 years.

关键词： Veterinary Anatomy Heart Preservation Techniques Alcohol Propylene Glycol Technique Fixation Necropsy Toxic Chemicals

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Phase equilibrium modelling of the amphibolite facies metamorphism in the Yelapa-Chimo Metamorphic Complex, Mexico

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Geoscience Frontiers 2021年第1期12卷 293-312页

作者： Fabian Gutierrez-Aguilar Peter Schaaf Gabriela Solís-pichardo Gerardo F.Arrieta-García Teodoro hernandez-Trevino Carlos Linares-Lopez Posgrado en Ciencias de la Tierra Instituto de GeofísicaUniversidad Nacional Autonoma de MexicoAv.Universidad 3000Ciudad de Mexico04510Mexico Laboratorio Universitario de Geoquímica Isotopica(LUGIS) Instituto de GeofísicaUniversidad Nacional Autonoma de MexicoAv.Universidad 3000Ciudad de Mexico04510Mexico Laboratorio Universitario de Geoquímica Isotopica(LUGIS) Instituto de GeologíaUniversidad Nacional Autonoma de MexicoAv.Universidad 3000Ciudad de Mexico04510Mexico Instituto de Geofísica Universidad Nacional Autonoma de MexicoAv.Universidad 3000Ciudad de Mexico04510Mexico

The Yelapa-Chimo Metamorphic Complex forms part of the jalisco Block in western Mexico and exposes a wide range of early Cretaceous metamorphic rocks;such as paragneiss,orthogneiss,amphibolites,and migmatites.However,the pressure-temperature(P-T)conditions of metamorphism and partial melting remain poorly studied in the region.To elucidate metamorphic P-T conditions,phase equilibrium modelling was applied to two sillimanite-garnet paragneisses,one amphibole-orthogneiss,and one amphibolite.Sillimanite-garnet paragneisses exhibit a lepidoblastic texture with a biotite+sillimanite+kyanite+garnet+quartz+plagioclase+K-feldspar mineral assemblage.Amphibole-orthogneiss and amphibolite display a nematoblastic texture with an amphibole+(1)plagioclase+quartz+(1)titanite assemblage and an amphibole+(2)plagioclase+(2)titanite+ilmenite retrograde mineral assemblage.Pseudosections calculated for the two sillimanite-garnet paragneiss samples show P-T peak conditions at~6-7.5 kbar and~725-740℃.The results for amphibole-orthogneiss and the amphibolite yield P-T peak conditions at~8.5-10 kbar and~690-710℃.The mode models imply that metasedimentary and metaigneous units can produce up to~20 vol%and~10 vol%of melt,respectively.Modelling within a closed system during isobaric heating suggests that melt compositions of metasedimentary and metaigneous units are likely to have direct implications for the petrogenesis of the Puerto Vallarta Batholith.Our new data indicate that the Yelapa-Chimo Metamorphic Complex evolved through a metamorphic gradient between~23-33℃km^-1and the metamorphic rocks formed at depths between~22 km and~30 km with a burial rate of~2.0 km Ma^-1.Finally,the P-T data for both metasedimentary and metaigneous rocks provide new constraints on an accretionary framework,which is responsible for generating metamorphism and partial melting in the YelapaChimo Metamorphic Complex during the early Cretaceous.

关键词： Phase equilibrium modelling Yelapa-Chimo Metamorphic Complex Amphibolite facies Partial melting

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effect of Different Reproductive Stages and Culture Times on Domestic Cat <i>in Vitro </i>Oocyte Maturation

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Advances in Reproductive Sciences 2021年第2期9卷 129-138页

作者： josé ernesto hernández pichardo Miguel Ramses Del Moral Reyes Michael e. Kjelland josé Luis Rodríguez Suastegui Department of Agricultural and Animal Production Division of Biological and Health SciencesMetropolitan Autonomous University-Xochimilco Mexico City Mexico Private Clinic Mexico City Mexico Conservation Genetics & Biotech LLC Valley City USA Biological and Health Sciences PhD Program Metropolitan Autonomous University-Xochimilco Mexico City Mexico

The domestic cat has been used as a model to carry out comparative research in assisted reproduction, to be applied in wild cats. The efficiency in domestic cat IVM concerning the reproductive status and/or cultivation times has previously been investigated;however, the studies were carried out separately. The objective of this research was to evaluate the maturation of oocytes of domestic cats of different reproductive stages using two different in vitro culture times. The ovaries were obtained by Ooforo-Salpingo-Hysterectomy of cats that were of the following groups: 1) prepubertal, 2) follicular, 3) pregnant or 4) in anestrus. Maturation was carried out with TCM199 medium supplemented with BSA for 24 h and 48 h. On average, 29 ± 25, 20 ± 15, 17 ± 9 and 17 ± 13 oocytes/cat were recovered from the prepubertal follicular, pregnant, and anestrus stages, respectively, but did not show a significant difference (P > 0.05). Also, meiotic maturation did not show a significant difference between the different reproductive stages at 24 h and 48 h, respectively (P > 0.05). However, in the prepubertal and follicular stages, greater oocyte maturation numbers were observed at 48 h compared to 24 h (P result was not observed in the pregnant and anestrus stages (P > 0.05), indicating that the in vitro culture duration is an important factor during in vitro maturation of domestic cat oocytes.

关键词： Domestic Cat Oocytes In Vitro Maturation Culture Time estrus Cycle Stages

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Use of IVF and eT in Mexican Criollo Sheep (Ovis aries): Immediate and Delayed embryo Transfers

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Advances in Reproductive Sciences 2016年第1期4卷 8-16页

作者： josé ernesto hernández pichardo Francisco Ortiz josé Luis Rodríguez Yvonne Ducolomb Filiberto Fernández Reyes Miguel Betancourt eduardo Casas Yvonne Heuze Michael e. Kjelland Salvador Romo Departamento de Producción Agrícola y Animal División de Ciencias Biológicas y de la Salud Universidad Autónoma Metropolitana-Xochimilco México DF México SVI México y Crio Vida Animal México Camino San Rafael 2 Col. San Rafael Querétaro México Departamento de Ciencias de la Salud División de Ciencias Biológicas y de la Salud Universidad Autónoma Metropolitana-Iztapalapa México DF México Conservation Genetics & Biotech LLC Valley City ND USA Laboratorio de Reproducción Facultad de Estudios Superiores Cuautitlán UNAM Estado de México México

There is little information in the scientific literature concerning sheep pregnancy and lambing success with regard to the timeframe from when in vitro produced embryos are transported to the designated location for embryo transfer (eT). The aim of this study was to transfer in vitro produced embryos under two different conditions that could typically occur using the aforementioned assisted reproductive techniques (ARTs). Abattoir ovaries were used to procure oocytes for in vitro embryo production and subsequent transfer to synchronized ewes. The study consisted of two experiments: experiment 1 (exp1)—embryos taken from the laboratory to a nearby surgical room for immediate eT, and experiment 2 (exp2)—eT after 5 hours (h) of transport to a rural farm. Lambing in relation to detected pregnancies, births compared to pregnancies, and the proportion of twin offspring were all higher in exp2. Notably, in both exp1 and exp2, there was not a significant difference (P > 0.05) between the number of embryos transferred, i.e., 3 versus 4, respectively, and the number of ewes that underwent parturition in each group. Also, in both experiments there was not a significant difference (P > 0.05) in the number of ewes that underwent parturition based on the number (i.e., ≥1) of corpora lutea present. The results of the present study demonstrate the importance of evaluating different conditions when applying ARTs, as there are many variables that can influence the outcome. Importantly, exp2 results show that ovine eT in places located far away from the embryo production site can be useful and successful provided that embryo transport, eT, and recipient conditions are adequate.

关键词： embryo (Transfer) Fertilization (In Vitro) Ovine Reproduction (Assisted) Sheep

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Raman spectroscopy for detection of imatinib in plasma: A proof of concept

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journal of Innovative Optical Health Sciences 2015年第5期8卷 56-66页

作者： Sanhita Rath Aditi Sahu Vikram Gota P.G.Martínez-Torres j.L.pichardo-Molina C.Murali Krishna Department of Clinical Pharmacology ACTREC Tata Memorial CenterKharghar Navi Mumbai 410210 India Chilakapati Lab ACTRECTata Memorial Center KhargharNavi Mumbai 410210 India Instituto de Fisica y Matematicas Universidad Michoacana de San Nicolas de Hidalgo edificio C-3 Ciudad Universitaria CP.58040 MoreliaMichoacanMexico Centro de Investigaciones enOptica AC C.P.37#150 Leon Gto.Mexico

Imatinib is the standard first line treatment for chronic myeloid leukemia(CML).Owing to doserelated toxicities of Imatinib such as neutropenia,there is scope for treatment optimization through therapeutic drug monitoring(TDM).Trough concentration of 1g/mL is considered the therapeutic threshhold.existing methods for the detection of Imatinib in plasma are limited by long read out time and expensive instrumentation.Hence,Raman spectroscopy was explored as a rapid and objective tool for monitoring Imatinib concentration.Three approaches:conventional Raman spectroscopy(CRS),Drop coating deposition Raman(DCDR)spectroscopy and surface-enhanced Raman spectroscopy(SeRS)were employed to detect the required trough concentration of 1g/mL and above.Detection of therapeutically relevant concentrations(1g/mL)using SeRS and suitable nanoparticle substrates has been demonstrated.Prospectively,rigorous validation using clinical samples is necessary to confirm the utility of this approach in routine clinical usage.

关键词： Therapeutic drug monitoring Raman spectroscopy Imatinib SeRS DCDR chronic myeloid leukemia

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High Intensity Focused Ultrasound Optimal Device Design for Targeted Prostate Cancer Treatment: A Numerical Study

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journal of Modern Physics 2013年第2期4卷 240-245页

作者： Laura Curiel Dexter Hobson jean-Yves Chapelon Samuel pichardo Lakehead University Thunder Bay Regional Research Institute Ontario Canada National Institute of Health and Medical Research (INSeRM) Lyon France

A parametric study was performed to design a device capable of treating small targeted regions within the prostate using high intensity focused ultrasound, while sparing the surrounding organs and minimizing the number of elements. The optimal focal length (L), operating frequency (f), element size (a) and central opening radius for lodging an imaging probe (r) of a device that would safely treat tissue within the prostate were obtained. Images from the Visible Human Project were used to determine simulated organ sizes and treatment locations. elliptical tumors were placed throughout the simulated prostate and their lateral and axial limits were selected as test locations. Using graphics processors, the acoustic field and Bio-Heat Transfer equation were solved to calculate the heating produced during a simulated treatment. L, f, a and r were varied from 45 to 75 mm, 2.25 to 3 MHz, 1.5 to 8 times the wavelength and 9 to 12.5 mm, respectively. The resulting optimal device was a 761-element concentric-ring transducer with L = 68 mm, f = 2.75 MHz, a = 2.05λ and r = 9 mm. Simulated thermal lesions showed that it was possible to treat target tumors consistent with reported locations and sizes for prostate cancer.

关键词： Focused Ultrasound Prostate Cancer Thermal Ablation Non-Invasive Treatment BHTe HIFU

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西班牙塞维利亚市男性同性恋患者中的性传播疾病

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地方病译丛 1992年第2期13卷 50-51页

作者： A R pichardo 牛新玲

过去10年人们越来越意识到,在有多个匿名性侣的男性同性恋者中患性传播疾病的危险增加,可能有症状,也可能无症状。在男性同性恋中发现的性病与异性恋者的类型有所不同。由于在西班牙除淋病和梅毒外均缺乏可靠的统计,我们进行了前瞻性的... 详细信息

过去10年人们越来越意识到,在有多个匿名性侣的男性同性恋者中患性传播疾病的危险增加,可能有症状,也可能无症状。在男性同性恋中发现的性病与异性恋者的类型有所不同。由于在西班牙除淋病和梅毒外均缺乏可靠的统计,我们进行了前瞻性的研究,以便确定两组到塞维利亚医学院性病诊所就诊的男性同性恋者的性病流行情况。该市人口为

关键词：男性同性恋异性恋同性恋者性病性传播疾病传染病淋球菌尿道炎患者塞维利亚西班牙南欧

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Low serum levels of ghrelin are associated with gallstone disease

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World journal of Gastroenterology 2006年第19期12卷 3096-3100页

作者： Nahum Méndez-Sánchez Guadalupe Ponciano-Rodríguez Luisa Bermejo-Martínez Antonio R Villa Norberto C Chávez-Tapia Daniel Zamora-Valdés Raúl pichardo-Bahena Blanca Barredo-Prieto Martha H Uribe-Ramos Martha H Ramos Héctor A Baptista-González Misael Uribe Liver Unit Biomedical Research Department Medica Sur Clinic and FoundationMexico CityMexico Faculty of Medicine National Autonomous University of Mexico (UNAM) Mexico CityMexico Clinical Research Department National Institute of Perinatology Mexico CityMexico Clinical epidemiology Unit National Institute of Medical Sciences and Nutrition Mexico CityMexico

瞄准：在胆石疾病探索 ghrelin 的角色。方法：我们在 150 个题目，执行了代表性的研究 38 与胆石(盒子) 和 112 控制。我们也各在二十件胆囊样品做了即时 PCR-RT 研究。身体集体索引(BMI ) ，浆液胰岛素， ghrelin，和浆液类脂化合物... 详细信息

瞄准：在胆石疾病探索 ghrelin 的角色。方法：我们在 150 个题目，执行了代表性的研究 38 与胆石(盒子) 和 112 控制。我们也各在二十件胆囊样品做了即时 PCR-RT 研究。身体集体索引(BMI ) ，浆液胰岛素， ghrelin，和浆液类脂化合物被测量。逻辑回归分析(univariate 并且多变量) 被进行估计与浆液 ghrelin 集中联系的胆石疾病的概率。结果：盒子与在性分发的控制统计上不同(P = 0.01 ) ，年龄(53 对 44 年， P = 0.002 ) ， BMI (28 对 25；P = 0.004 ) ，并且葡萄糖(5.26 对 4.98 mmol/L；P = 0.05 ) 。没有新陈代谢的症候群，在第三 tercile 上面的 ghrelin 浆液层次的流行在题目是更低的(P elin 价值比中部的价值高时，我们发现了保护的效果(或 = 0.27， 95%CI 0.09-0.82， P = 0.02 ) 。二十(20%) 胆囊标本表示了 ghrelin mRNA。结论：浆液 ghrelin 集中与 GD 的保护的效果被联系。

关键词：血液水平胆结石肥胖胆囊

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天生慈母心

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疯狂英语（阅读版） 2005年第10期 34-37页

作者： Winifred pichardo 柏柏

回首走过的生命历程,我不禁想问,我都做了些什么重要或有意义的事情,我取得了哪些方面的成就呢?我对身边的人,尤其是心爱的那些人,都产生了一些什么影响或者让他们收获到了什么呢?

关键词：生命历程成就收获意义母亲交流照顾重要方面新生活

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