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1.
Animals (Basel) ; 14(7)2024 Mar 29.
Article in English | MEDLINE | ID: mdl-38612280

ABSTRACT

Pre-implantation embryos release extracellular vesicles containing different molecules, including DNA. The presence of embryonic DNA in E-EVs released into the culture medium during in vitro embryo production could be useful for genetic diagnosis. However, the vesicles containing DNA might be derived from embryos suffering from apoptosis, i.e., embryos of bad quality. This work intended to confirm that embryos release DNA that is useful for genotyping by evaluating the effect of embryonic apoptosis on DNA content in E-EVs. Bovine embryos were produced by parthenogenesis and in vitro fertilization (IVF). On Day 5, morulae were transferred to individual cultures in an EV-depleted SOF medium. On Day 7, embryos were used to evaluate cellular apoptosis, and each culture medium was collected to evaluate E-EV concentration, characterization, and DNA quantification. While no effect of the origin of the embryo on the apoptotic rate was found, arrested morulae had a higher apoptotic rate. E-EVs containing DNA were identified in all samples, and the concentration of those vesicles was not affected by the origin or quality of the embryos. However, the concentration of DNA was higher in EVs released by the arrested parthenogenetic embryos. There was a correlation between the concentration of E-EVs, the concentration of DNA-positive E-EVs, and the concentration of DNA. There was no negative effect of apoptotic rate on DNA-positive E-EVs and DNA concentration; however, embryos of the best quality with a low apoptotic rate still released EVs containing DNA. This study confirms that the presence of DNA in E-EVs is independent of embryo quality. Therefore, E-EVs could be used in liquid biopsy for noninvasive genetic diagnosis.

2.
Int J Mol Sci ; 25(8)2024 Apr 14.
Article in English | MEDLINE | ID: mdl-38673927

ABSTRACT

Domestic cat blastocysts cultured without the zona pellucida exhibit reduced implantation capacity. However, the protein expression profile has not been evaluated in these embryos. The objective of this study was to evaluate the protein expression profile of domestic cat blastocysts cultured without the zona pellucida. Two experimental groups were generated: (1) domestic cat embryos generated by IVF and cultured in vitro (zona intact, (ZI)) and (2) domestic cat embryos cultured in vitro without the zona pellucida (zona-free (ZF group)). The cleavage, morula, and blastocyst rates were estimated at days 2, 5 and 7, respectively. Day 7 blastocysts and their culture media were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The UniProt Felis catus database was used to identify the standard proteome. No significant differences were found in the cleavage, morula, or blastocyst rates between the ZI and ZF groups (p > 0.05). Proteomic analysis revealed 22 upregulated and 20 downregulated proteins in the ZF blastocysts. Furthermore, 14 proteins involved in embryo development and implantation were present exclusively in the culture medium of the ZI blastocysts. In conclusion, embryo culture without the zona pellucida did not affect in vitro development, but altered the protein expression profile and release of domestic cat blastocysts.


Subject(s)
Blastocyst , Proteomics , Zona Pellucida , Animals , Blastocyst/metabolism , Zona Pellucida/metabolism , Cats , Proteomics/methods , Embryo Culture Techniques , Secretome/metabolism , Female , Fertilization in Vitro , Proteome/metabolism , Embryonic Development , Tandem Mass Spectrometry , Chromatography, Liquid
3.
Int J Mol Sci ; 24(8)2023 Apr 18.
Article in English | MEDLINE | ID: mdl-37108601

ABSTRACT

The embryo-maternal interaction occurs during the early stages of embryo development and is essential for the implantation and full-term development of the embryo. In bovines, the secretion of interferon Tau (IFNT) during elongation is the main signal for pregnancy recognition, but its expression starts around the blastocyst stage. Embryos release extracellular vesicles (EVs) as an alternative mechanism of embryo-maternal communication. The aim of the study was to determine whether EVs secreted by bovine embryos during blastulation (D5-D7) could induce transcriptomic modifications, activating IFNT signaling in endometrial cells. Additionally, it aims to assess whether the EVs secreted by embryos produced in vivo (EVs-IVV) or in vitro (EVs-IVP) have different effects on the transcriptomic profiles of the endometrial cells. In vitro- and in vivo-produced bovine morulae were selected and individually cultured for 48 h to collect embryonic EVs (E-EVs) secreted during blastulation. E-EVs stained with PKH67 were added to in vitro-cultured bovine endometrial cells to assess EV internalization. The effect of EVs on the transcriptomic profile of endometrial cells was determined by RNA sequencing. EVs from both types of embryos induced several classical and non-classical IFNT-stimulated genes (ISGs) and other pathways related to endometrial function in epithelial endometrial cells. Higher numbers of differentially expressed genes (3552) were induced by EVs released by IVP embryos compared to EVs from IVV (1838). Gene ontology analysis showed that EVs-IVP/IVV induced the upregulation of the extracellular exosome pathway, the cellular response to stimulus, and the protein modification processes. This work provides evidence regarding the effect of embryo origin (in vivo or in vitro) on the early embryo-maternal interaction mediated by extracellular vesicles.


Subject(s)
Embryo, Mammalian , Extracellular Vesicles , Animals , Cattle , Female , Pregnancy , Blastocyst/metabolism , Embryo, Mammalian/metabolism , Embryonic Development/genetics , Endometrium , Extracellular Vesicles/metabolism , Parturition , Interferons/metabolism
4.
Reprod Biol ; 23(1): 100725, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36565511

ABSTRACT

Pre-implantation embryos release extracellular vesicles (EVs) to extracellular environment. In this work it is hypothesized that the EVs miRNA cargo will vary during pre-implantation development due to the constant changes in gene expression that take place through this period. The concentration, size and miRNA cargo of EVs secreted by competent bovine embryos during the period from compaction to blastulation (Day 3-7) were analyzed. For this analysis tow developmental windows were defined: W2 from 8-cells (D3) to morula (D5) and W3 from morula (D5) to blastocyst (D7). For W2, in vitro produced embryos were individually cultured in EVs-depleted medium from D3 to D5; culture media were collected and assigned to Group W2. Morulae were kept in culture up to blastocyst stage to determine the developmental competence. For W3, D5 morulae were collected and cultured individually in EVs-depleted medium up to blastocyst stage; culture media were assigned to Group W3, and blastocysts were kept in culture up to day 11 to define their competence. The mean size of EVs was similar between groups, however, EVs concentration was lower in W2. A total of 140 miRNAs were identified. From them, 79 were differentially expressed between the groups, 28 upregulated and 51 downregulated. miRNAs differentially detected between both developmental windows participate in the regulation of signaling pathways which crucial for embryonic development. It was concluded that the secretion of EVs is regulated by the developmental progress of the embryo during the pre-implantation period.


Subject(s)
Extracellular Vesicles , MicroRNAs , Pregnancy , Female , Animals , Cattle , MicroRNAs/metabolism , Embryo Culture Techniques , Embryo Implantation , Blastocyst/physiology , Extracellular Vesicles/genetics , Extracellular Vesicles/metabolism , Culture Media
5.
Rev Panam Salud Publica ; 46: e132, 2022.
Article in Spanish | MEDLINE | ID: mdl-36071918

ABSTRACT

Objective: To describe the types of blood pressure measurement devices marketed through the Mercado Público platform between 2018 and 2020. Methods: This is a descriptive study based on transactions on the Mercado Público platform for the purchase of blood pressure measurement devices in the 2018-2020 period. A search was made for any validation granted by international organizations for automatic devices. The results are expressed as absolute and relative frequencies. Results: Between 2018 and 2020, purchases of automatic devices increased from 15% to 75%, while purchases of mercury-based devices dropped from 40.9% to 7.5%, and aneroid devices fell from 38.7% to 15.8%. Of the 772 automatic devices, 52.6% were validated for accuracy. Conclusions: Given that Chile is committed to the HEARTS Initiative in the Americas, and that prevention and control of hypertension is critical to reducing mortality from noncommunicable diseases, it is crucial to ensure accurate blood pressure measurement. This document provides relevant information on transactions and the purchase profile of blood pressure measurement devices on the Mercado Público platform. A growing trend was observed in the purchase of validated automatic equipment during the evaluation period.


Objetivo: Descrever os tipos de dispositivos de aferição da pressão arterial comercializados por meio da plataforma Mercado Público, entre 2018 e 2020. Métodos: Estudo descritivo das transações realizadas por meio do Mercado Público, no período de 2018 a 2020, para a compra de dispositivos de aferição da pressão arterial. No caso de dispositivos automáticos, foi verificado se possuíam validação concedida por organizações internacionais. Os resultados são apresentados na forma de frequências absolutas e relativas. Resultados: Entre 2018 e 2020, a aquisição de dispositivos automáticos aumentou de 15% para 75%. Houve uma redução de 40,9% para 7,5% na compra de esfigmomanômetros de mercúrio e de 38,7% para 15,8% na compra de esfigmomanômetros aneroides. Dos 772 dispositivos automáticos, 52,6% tinham exatidão validada. Conclusões: Considerando que o Chile está comprometido com a Iniciativa HEARTS nas Américas e que a prevenção e o controle da hipertensão arterial sistêmica é fundamental para reduzir a mortalidade por doenças não transmissíveis, é essencial garantir a aferição exata da pressão arterial. Este documento fornece informações relevantes sobre as transações realizadas e o perfil de compra de dispositivos de aferição da pressão arterial por meio da plataforma Mercado Público, mostrando uma tendência crescente na aquisição de dispositivos automáticos validados durante o período de avaliação.

6.
Rev Panam Salud Publica ; 46, 2022. Special Issue HEARTS
Article in Spanish | PAHO-IRIS | ID: phr-56326

ABSTRACT

[RESUMEN]. Objetivo. Describir los tipos de dispositivos de medición de presión arterial comercializados a través de la plataforma Mercado Público entre los años 2018 y 2020. Métodos. Estudio de tipo descriptivo procedente de las transacciones realizadas a través de Mercado Público en el periodo 2018 y 2020, para la adquisición de dispositivos de medición de presión arterial. Para los equi- pos automáticos, se buscó el estado de validación otorgado por organismos internacionales. Los resultados se presentan en frecuencias absolutas y relativas. Resultados. Entre los años 2018-2020, aumentó la adquisición de dispositivos automáticos de un 15% a un 75%, y se presentó una reducción del 40,9% al 7,5% en equipos mercuriales y de 38,7% al 15,8% en equipos aneroides. De los 772 dispositivos automáticos, el 52,6% contó con validación de precisión. Conclusiones. Considerando que Chile está comprometido con la Iniciativa HEARTS en las Américas, y que la prevención y control de la HTA es fundamental para reducir la mortalidad por enfermedades no trasmisibles, resulta crucial garantizar una medición de presión arterial precisa. El presente documento aporta información relevante respecto de las transacciones realizadas y el perfil de compra de dispositivos de medición de presión arterial a través de la plataforma Mercado Público, observándose una tendencia creciente en la obtención de equipos automáticos validados durante el periodo evaluado.


[ABSTRACT]. Objective. To describe the types of blood pressure measurement devices marketed through the Mercado Público platform between 2018 and 2020. Methods. This is a descriptive study based on transactions on the Mercado Público platform for the purchase of blood pressure measurement devices in the 2018-2020 period. A search was made for any validation granted by international organizations for automatic devices. The results are expressed as absolute and rela- tive frequencies. Results. Between 2018 and 2020, purchases of automatic devices increased from 15% to 75%, while pur- chases of mercury-based devices dropped from 40.9% to 7.5%, and aneroid devices fell from 38.7% to 15.8%. Of the 772 automatic devices, 52.6% were validated for accuracy. Conclusions. Given that Chile is committed to the HEARTS Initiative in the Americas, and that prevention and control of hypertension is critical to reducing mortality from noncommunicable diseases, it is crucial to ensure accurate blood pressure measurement. This document provides relevant information on transactions and the purchase profile of blood pressure measurement devices on the Mercado Público platform. A growing trend was observed in the purchase of validated automatic equipment during the evaluation period.


[RESUMO]. Objetivo. Descrever os tipos de dispositivos de aferição da pressão arterial comercializados por meio da plataforma Mercado Público, entre 2018 e 2020. Métodos. Estudo descritivo das transações realizadas por meio do Mercado Público, no período de 2018 a 2020, para a compra de dispositivos de aferição da pressão arterial. No caso de dispositivos automáticos, foi verificado se possuíam validação concedida por organizações internacionais. Os resultados são apresenta- dos na forma de frequências absolutas e relativas. Resultados. Entre 2018 e 2020, a aquisição de dispositivos automáticos aumentou de 15% para 75%. Houve uma redução de 40,9% para 7,5% na compra de esfigmomanômetros de mercúrio e de 38,7% para 15,8% na compra de esfigmomanômetros aneroides. Dos 772 dispositivos automáticos, 52,6% tinham exatidão validada. Conclusões. Considerando que o Chile está comprometido com a Iniciativa HEARTS nas Américas e que a prevenção e o controle da hipertensão arterial sistêmica é fundamental para reduzir a mortalidade por doenças não transmissíveis, é essencial garantir a aferição exata da pressão arterial. Este documento fornece informações relevantes sobre as transações realizadas e o perfil de compra de dispositivos de aferição da pressão arterial por meio da plataforma Mercado Público, mostrando uma tendência crescente na aquisição de dispositivos automáticos validados durante o período de avaliação.


Subject(s)
Sphygmomanometers , Arterial Pressure , Hypertension , Chile , Sphygmomanometers , Arterial Pressure , Hypertension , Sphygmomanometers , Arterial Pressure , Hypertension
7.
Animals (Basel) ; 12(2)2022 Jan 09.
Article in English | MEDLINE | ID: mdl-35049777

ABSTRACT

Over the last few years, several commercial FSH products have been developed for cattle superovulation (SOV) purposes in Multiple Ovulation and Embryo Transfer (MOET) programs. The SOV response is highly variable among individuals and remains one of the main limiting factors in obtaining a profitable number of transferable embryos. In this study, follicle stimulating hormone (FSH) from different origins was included in two SOV protocols, (a) FSH from purified pig pituitary extract (NIH-FSH-p; two doses/day, 12 h apart, four consecutive days); and (b) extra-long-acting bovine recombinant FSH (bscrFSH; a single dose/day, four consecutive days), to test the effects of bscrFSH on the ovarian response, hormone profile levels, in vivo embryo production and the pluripotency gene expression of the obtained embryos. A total of 68 healthy primiparous red Angus cows (Bos taurus) were randomly distributed into two experimental groups (n = 34 each). Blood sample collection for progesterone (P4) and cortisol (C) level determination was performed together with ultrasonographic assessment for ovarian size, follicles (FL) and corpora lutea (CL) quantification in each SOV protocol (Day 0, 4, 8, and 15). Moreover, FSH profiles were monitorised throughout both protocols (Day 0, 4, 5, 6, 7, 8, 9, 10, and 15). In vivo embryo quantity and quality (total structures, morulae, blastocysts, viable, degenerated and blocked embryos) were recorded in each SOV protocol. Finally, embryo quality in both protocols was assessed by the analysis of the expression level of crucial genes for early embryo development (OCT4, IFNt, CDX2, BCL2, and BAX). P4 and cortisol concentration peaks in both SOV protocols were obtained on Day 15 and Day 8, respectively, which were statistically different compared to the other time-points (p < 0.05). Ovarian dimensions increased from Day 0 to Day 15 irrespective of the SOV protocol considered (p < 0.05). Significant changes in CL number were observed over time till Day 15 irrespective of the SOV protocol applied (p < 0.05), being non- significantly different between SOV protocols within each time-point (p > 0.05). The number of CL was higher on Day 15 in the bscrFSH group compared to the NIH-FSH-p group (p < 0.05). The number of embryonic structures recovered was higher in the bscrFSH group (p = 0.025), probably as a result of a tendency towards a greater number of follicles developed compared to the NIH-FSH-p group. IFNt and BAX were overexpressed in embryos from the bscrFSH group (p < 0.05), with a fold change of 16 and 1.3, respectively. However, no statistical differences were detected regarding the OCT4, CDX2, BCL2, and BCL2/BAX expression ratio (p > 0.05). In conclusion, including bscrFSH in SOV protocols could be an important alternative by reducing the number of applications and offering an improved ovarian response together with better embryo quality and superior performance in embryo production compared to NIH-FSH-p SOV protocols.

8.
Rev. panam. salud pública ; 46: e132, 2022. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1431964

ABSTRACT

RESUMEN Objetivo. Describir los tipos de dispositivos de medición de presión arterial comercializados a través de la plataforma Mercado Público entre los años 2018 y 2020. Métodos. Estudio de tipo descriptivo procedente de las transacciones realizadas a través de Mercado Público en el periodo 2018 y 2020, para la adquisición de dispositivos de medición de presión arterial. Para los equipos automáticos, se buscó el estado de validación otorgado por organismos internacionales. Los resultados se presentan en frecuencias absolutas y relativas. Resultados. Entre los años 2018-2020, aumentó la adquisición de dispositivos automáticos de un 15% a un 75%, y se presentó una reducción del 40,9% al 7,5% en equipos mercuriales y de 38,7% al 15,8% en equipos aneroides. De los 772 dispositivos automáticos, el 52,6% contó con validación de precisión. Conclusiones. Considerando que Chile está comprometido con la Iniciativa HEARTS en las Américas, y que la prevención y control de la HTA es fundamental para reducir la mortalidad por enfermedades no trasmisibles, resulta crucial garantizar una medición de presión arterial precisa. El presente documento aporta información relevante respecto de las transacciones realizadas y el perfil de compra de dispositivos de medición de presión arterial a través de la plataforma Mercado Público, observándose una tendencia creciente en la obtención de equipos automáticos validados durante el periodo evaluado.


ABSTRACT Objective. To describe the types of blood pressure measurement devices marketed through the Mercado Público platform between 2018 and 2020. Methods. This is a descriptive study based on transactions on the Mercado Público platform for the purchase of blood pressure measurement devices in the 2018-2020 period. A search was made for any validation granted by international organizations for automatic devices. The results are expressed as absolute and relative frequencies. Results. Between 2018 and 2020, purchases of automatic devices increased from 15% to 75%, while purchases of mercury-based devices dropped from 40.9% to 7.5%, and aneroid devices fell from 38.7% to 15.8%. Of the 772 automatic devices, 52.6% were validated for accuracy. Conclusions. Given that Chile is committed to the HEARTS Initiative in the Americas, and that prevention and control of hypertension is critical to reducing mortality from noncommunicable diseases, it is crucial to ensure accurate blood pressure measurement. This document provides relevant information on transactions and the purchase profile of blood pressure measurement devices on the Mercado Público platform. A growing trend was observed in the purchase of validated automatic equipment during the evaluation period.


RESUMO Objetivo. Descrever os tipos de dispositivos de aferição da pressão arterial comercializados por meio da plataforma Mercado Público, entre 2018 e 2020. Métodos. Estudo descritivo das transações realizadas por meio do Mercado Público, no período de 2018 a 2020, para a compra de dispositivos de aferição da pressão arterial. No caso de dispositivos automáticos, foi verificado se possuíam validação concedida por organizações internacionais. Os resultados são apresentados na forma de frequências absolutas e relativas. Resultados. Entre 2018 e 2020, a aquisição de dispositivos automáticos aumentou de 15% para 75%. Houve uma redução de 40,9% para 7,5% na compra de esfigmomanômetros de mercúrio e de 38,7% para 15,8% na compra de esfigmomanômetros aneroides. Dos 772 dispositivos automáticos, 52,6% tinham exatidão validada. Conclusões. Considerando que o Chile está comprometido com a Iniciativa HEARTS nas Américas e que a prevenção e o controle da hipertensão arterial sistêmica é fundamental para reduzir a mortalidade por doenças não transmissíveis, é essencial garantir a aferição exata da pressão arterial. Este documento fornece informações relevantes sobre as transações realizadas e o perfil de compra de dispositivos de aferição da pressão arterial por meio da plataforma Mercado Público, mostrando uma tendência crescente na aquisição de dispositivos automáticos validados durante o período de avaliação.

9.
Zygote ; 29(2): 138-149, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33118919

ABSTRACT

Human embryos generated in vitro have a high incidence of chromosomal abnormalities that negatively affect pregnancy rate. Embryos generated in vitro secrete extracellular vesicles (EVs) into the culture medium that could be used potentially as indicators of embryo competence. This research aimed to evaluate the concentration and size of EVs and their gDNA content as an indicator of developmental competence in human embryos. Human embryos generated by intracytoplasmic sperm injection (ICSI) were classified morphologically as of either TOP, FAIR or POOR quality. Culture medium and developmentally arrested embryos (which were not able to be used for embryo transfer) were collected. Microvesicles, exosomes (MV/Exo) and apoptotic bodies (ABs) were isolated from culture medium. Nanoparticle tracking analysis (NTA) and array comparative genomic hybridization (aCGH) analysis were performed to evaluate EVs and their gDNA content. From NTA, the diameter (mean) of MVs/Exo from TOP quality embryos was higher (112.17 nm) compared with that of FAIR (108.02) and POOR quality embryos (102.78 nm) (P < 0.05). aCGH analysis indicated that MVs/Exo and ABs carried gDNA with the presence of 23 chromosome pairs. However, when arrested embryos were compared with their respective MVs/Exo and ABs, the latter had an increased rate of chromosomal abnormalities (24.9%) compared with embryos (8.7%) (P < 0.05). In conclusion, the size of EVs from culture medium might be an alternative for evaluating competence of human embryos, however more studies are needed to validate the use of gDNA from EVs as an indicator of embryo competence.


Subject(s)
Embryo Culture Techniques , Extracellular Vesicles , Blastocyst , Comparative Genomic Hybridization , Embryo, Mammalian , Humans
10.
Int J Mol Sci ; 21(23)2020 Nov 24.
Article in English | MEDLINE | ID: mdl-33255183

ABSTRACT

During early development, embryos secrete extracellular vesicles (EVs) that participate in embryo-maternal communication. Among other molecules, EVs carry microRNAs (miRNAs) that interfere with gene expression in target cells; miRNAs participate in embryo-maternal communication. Embryo selection based on secreted miRNAs may have an impact on bovine breeding programs. This research aimed to evaluate the size, concentration, and miRNA content of EVs secreted by bovine embryos with different developmental potential, during the compaction period (days 3.5-5). Individual culture media from in vitro-produced embryos were collected at day 5, while embryos were further cultured and classified at day 7, as G1 (conditioned-culture media by embryos arrested in the 8-16-cells stage) and G2 (conditioned-culture media by embryos that reached blastocyst stages at day 7). Collected nanoparticles from embryo conditioned culture media were cataloged as EVs by their morphology and the presence of classical molecular markers. Size and concentration of EVs from G1 were higher than EVs secreted by G2. We identified 95 miRNAs; bta-miR-103, bta-miR-502a, bta-miR-100, and bta-miR-1 were upregulated in G1, whereas bta-miR-92a, bta-miR-140, bta-miR-2285a, and bta-miR-222 were downregulated. The most significant upregulated pathways were fatty acid biosynthesis and metabolism, lysine degradation, gap junction, and signaling pathways regulating pluripotency of stem cells. The characteristics of EVs secreted by bovine embryos during the compaction period vary according to embryo competence. Embryos that reach the blastocyst stage secrete fewer and smaller vesicles. Furthermore, the loading of specific miRNAs into the EVs depends on embryo developmental competence.


Subject(s)
Biomarkers/metabolism , Embryonic Development/genetics , Extracellular Vesicles/genetics , MicroRNAs/genetics , Animals , Blastocyst/metabolism , Cattle , Culture Media, Conditioned/pharmacology , Embryo Culture Techniques , Embryo, Mammalian , Female
11.
Theriogenology ; 158: 148-157, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32961350

ABSTRACT

The kodkod (Leopardus guigna) is a small felid endemic of Chile and is considered a vulnerable species. Domestic cat oocytes have been successfully used as recipient cytoplast to reprogram somatic cells from different felids by interspecific somatic cell nuclear transfer (iSCNT). The developmental competence of felid embryos generated by iSCNT can be improved by the aggregation method using a zona-free culture system. The objective of this research was to evaluate the developmental competence of kodkod embryos generated by iSCNT using domestic cat oocytes and the aggregation method. For this purpose, five experimental group were done: (1) cat embryos generated by IVF, (2) cat embryos generated by SCNT (Ca1x), (3) aggregated cat embryos generated by SCNT (Ca2x), (4) kodkod embryos generated by iSCNT (K1x) and (5) aggregated kodkod embryos generated by iSCNT (K2x). Cleavage, morulae and blastocyst rates were estimated. The blastocyst diameter was evaluated. The gene expression level of pluripotency (OCT4, SOX2 and NANOG) and differentiation markers (CDX2 and GATA6) was analyzed in blastocysts. Morulae rate was higher in the IVF group and when cloned embryos were cultured in aggregates (IVF: 68.2%, Ca2x: 58.0% and K2x: 62.4%) compared to individually cultured kodkod embryos (K1x: 37.0%) (P < 0.05). Embryo aggregation increased blastocysts formation in the Ca2x group (30.9%) to a similar rate compared to the IVF group (44.5%) (P > 0.05). No blastocysts were generated in the K1x group, whereas blastocysts formation was obtained in K2x group (5.9%). The diameter of blastocysts from the K2x group (172.8 µm) was significantly lower than blastocysts from the Ca2x group (P < 0.05). The relative expression of OCT4 was lower in blastocysts from Ca1x than in blastocysts from IVF (P < 0.05). Furthermore, CDX2 expression was lower in blastocysts from Ca2x than in blastocysts from Ca1x and IVF groups (P < 0.05). In kodkod embryos, only one blastocyst from the K2x group expressed OCT4. No expression of SOX2, NANOG, CDX2 and GATA6 was detected in kodkod blastocysts. In conclusion, after iSCNT, domestic cat oocytes support the development of kodkod embryos until the morula stage. The aggregation method increases the morulae rate of kodkod cloned embryos and allows blastocysts formation. However, kodkod blastocysts have a poor morphological quality and a lacking expression of pluripotency and differentiation markers, probably caused by an incomplete nuclear reprogramming.


Subject(s)
Blastocyst , Embryo, Mammalian , Animals , Cats , Chile , Cloning, Organism/veterinary , Embryonic Development , Nuclear Transfer Techniques/veterinary
12.
Anim Reprod ; 17(2): e20190109, 2020 May 05.
Article in English | MEDLINE | ID: mdl-32714450

ABSTRACT

Adipose derived mesenchymal stem cells (AMSCs) have been isolated from domestic and wild cats. For wild cats, the isolation of AMSCs has been reported in the black-footed cats (Felis nigripes) and guigna (Leopardus guigna). Stromal vascular fraction (SVF) isolated from cougar adipose tissue have been used to restore elbow functionality in the cougar (Puma concolor) but multipotent characteristics of these cells have not been described. The present study describes for the first time the isolation and characterization of mesenchymal stem cells derived from adipose tissue of cougar. AMSCs and fibroblasts from six months female cougar were isolated and cultured in DMEM/F12, supplemented with FBS 10% + 1% Antibiotic/Antifungal + 2.4 mM L-Glutamine + 2.4 mM pyruvate up to passage 5. Expression of pluripotent and surface marker genes was evaluated at mRNA level. Mesodermal differentiation (adipogenic, osteogenic and chondrogenic) was described. AMSCs expressed mRNA of pluripotent genes Oct4, Nanog, Sox2 and Klf4 and surface markers Cd44, Cd90, Cd105 and MHCII. Fibroblasts showed similar mRNA expression with the exception of Sox2. AMSCs obtained from cougar exhibit multipotency features similar to domestic cats MSC, nevertheless, other analyses are required. AMSCs from cougar could be a source of interest for treatment of individuals that remain in captivity or arrive to wildlife rehabilitation centers.

13.
Rev Med Chil ; 147(4): 470-474, 2019 Apr.
Article in Spanish | MEDLINE | ID: mdl-31344209

ABSTRACT

Obesity is a global health problem. Its worldwide prevalence has tripled between 1975 and 2016, reaching a prevalence in Chile of 34.4%, according to the National Health Survey 2016-2017. If this condition corresponds to a risk factor or primary disease is a widely discussed issue. It is recognized as a disease by the American Medical Association and World Health Organization, based on its metabolic and hormonal features, such as dysregulation of appetite, abnormal energy balance and endocrine dysfunction, among others. Its main environmental risk factors are the consumption of ultra-processed foods and sedentariness. Preventive measures at the population level are fundamental, emphasizing promotion and prevention using a transdisciplinary approach. The individual approach in the management of obesity should improve the quality of life, avoid early mortality, reduce cardiovascular risk, and reduce the progression to type 2 diabetes and incidence of cancer. Thus, an adequate management and control of obesity would have a great impact in our society.


Subject(s)
Obesity/complications , Cardiovascular Diseases/etiology , Diabetes Mellitus/etiology , Humans , Metabolic Diseases/etiology , Obesity/physiopathology , Risk Factors
14.
Rev. méd. Chile ; 147(4): 470-474, abr. 2019. graf
Article in Spanish | LILACS | ID: biblio-1014248

ABSTRACT

Obesity is a global health problem. Its worldwide prevalence has tripled between 1975 and 2016, reaching a prevalence in Chile of 34.4%, according to the National Health Survey 2016-2017. If this condition corresponds to a risk factor or primary disease is a widely discussed issue. It is recognized as a disease by the American Medical Association and World Health Organization, based on its metabolic and hormonal features, such as dysregulation of appetite, abnormal energy balance and endocrine dysfunction, among others. Its main environmental risk factors are the consumption of ultra-processed foods and sedentariness. Preventive measures at the population level are fundamental, emphasizing promotion and prevention using a transdisciplinary approach. The individual approach in the management of obesity should improve the quality of life, avoid early mortality, reduce cardiovascular risk, and reduce the progression to type 2 diabetes and incidence of cancer. Thus, an adequate management and control of obesity would have a great impact in our society.


Subject(s)
Humans , Obesity/complications , Cardiovascular Diseases/etiology , Risk Factors , Diabetes Mellitus/etiology , Metabolic Diseases/etiology , Obesity/physiopathology
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