Estudio Histológico en Embriones de Cobayo (Cavia porcellus) y su Utilidad como Modelo para la Comprensión del Desarrollo Embrionario Humano / Histological Study in Guinea Pig Embryos (Cavia porcellus) and its Usefulness as a Model for Understanding Human Embryonic Development

El cobayo es un modelo animal ampliamente utilizado en la investigación biomédica debido a sus similitudes biológicas con los seres humanos. El objetivo de nuestro estudio es proporcionar sustento morfológico para utilizar preparados histológicos de embriones de cobayo como modelo de estudio para comprender los procesos del desarrollo embrionario humano. Nuestros resultados muestran que los embriones de cobayo presentan características morfológicas similares a las observadas en los embriones humanos, lo que sugiere que pueden utilizarse como un modelo efectivo para estudiar el desarrollo embrionario humano. Este hallazgo tiene importantes implicancias para la investigación y la docencia utilizando este modelo animal. Se analizaron preparados histológicos de embriones de cobayo teñidos con hematoxilina eosina, adquiridos por la Universidad Autónoma de Chile. Se tomaron microfotografías de preparados histológicos de cobayo en diferentes estadios del desarrollo y se seleccionaron las mejores imágenes para la descripción de estructuras y establecer estimados de la embriogénesis. Del análisis de los preparados se desprende que órganos como esófago, médula espinal y corazón presentan similitudes anatómicas e histológicas que hacen posible compararlas con el desarrollo embrionario humano y la edad de gestación en etapas tempranas. El uso de preparados de embriones de cobayo y su análisis desde un aspecto histológico resulta ser una estrategia metodológica factible debido a las similitudes en la embriogénesis de los mamíferos y las concordancias morfológicas con el desarrollo de los órganos entre humanos y roedores. Esto permite implementar este modelo animal como una herramienta para comprender el desarrollo embrionario humano.

SUMMARY: The guinea pig is an animal model widely used in biomedical research due to its biological similarities with humans. The objective of our study is to provide morphological support to use histological preparations of guinea pig embryos as a study model to understand the processes of human embryonic development. Our results show that guinea pig embryos present morphological characteristics similar to those observed in human embryos, suggesting that they can be used as an effective model to study human embryonic development. This finding has important implications for research and teaching using this animal model. Histological preparations of guinea pig embryos stained with hematoxylin eosin, acquired by the Autonomous University of Chile, were analyzed. Photomicrographs of histological preparations of guinea pigs at different stages of development were taken and the best images were selected to describe structures and establish estimates of embryogenesis. From the analysis of the preparations it is clear that organs such as the esophagus, spinal cord and heart present anatomical and histological similarities that make it possible to compare them with human embryonic development and gestation age in early stages. The use of guinea pig embryo preparations and their analysis from a histological aspect turns out to be a feasible methodological strategy due to the similarities in mammalian embryogenesis and the morphological concordances with the development of organs between humans and rodents. This allows this animal model to be implemented as a tool to understand human embryonic development.

Teratoma mesentérico maduro en una mujer adulta: reporte de caso y revisión de la literatura / Mature mesenteric teratoma in an adult women: Case report and literature

Introducción. Los teratomas son neoplasias que surgen a partir de células germinales pluripotenciales y derivan de dos o más capas de células. Se clasifican en tumores maduros, que contienen tejidos bien diferenciados, o inmaduros, que contienen estructuras inmaduras y embrionarias. Su localización más frecuente son las gónadas; la ubicación mesentérica es infrecuente y se han descrito aproximadamente 40 casos en la literatura mundial. Dentro del abordaje diagnóstico y terapéutico, se emplea la tomografía computarizada y la resonancia magnética nuclear para caracterizar la lesión, evaluar la extensión intraabdominal y la relación con otras estructuras. El diagnóstico debe confirmarse mediante el examen histopatológico. Caso clínico. Paciente femenina de 56 años, con antecedente de carcinoma ductal infiltrante de mama izquierda en remisión, en estudios de seguimiento con hallazgo incidental en tomografía de abdomen de lesión abdominopélvica dependiente del mesenterio, contornos lisos y nivel grasa-líquido. Estudios de extensión con marcadores tumorales negativos. Resultados. Por la alta sospecha clínica e imagenológica de teratoma, fue llevada a resección quirúrgica de la lesión. El examen histopatológico confirmó el diagnóstico de teratoma quístico maduro del mesenterio. Conclusión. El teratoma mesentérico es una entidad clínica rara, que debe ser considerado como uno de los diagnósticos diferenciales de una masa abdominal con efecto compresivo. El diagnóstico se basa principalmente en el examen clínico y los hallazgos imagenológicos. La escisión quirúrgica temprana es el pilar del tratamiento; el abordaje laparoscópico o abierto depende de las características clínicas y la experiencia del cirujano.

Background. Teratomas are neoplasms that arise from pluripotent germ cells, derived from two or more layers of germ cells. They are classified as mature tumors (cystic or solid), which contain well-differentiated tissues, or as immature tumors, which contain immature and embryonic structures. Its most frequent location is the female and male gonads; the mesenteric location is rare and approximately 40 cases have been described in the world literature. Within the diagnostic and therapeutic approach, computed tomography and magnetic resonance imaging are used to characterize the lesion, assess intra-abdominal extension and the relationship with other structures. The diagnosis must be confirmed by histopathological examination. Clinical case. A 56-year-old female patient with a history of infiltrating ductal carcinoma of the left breast in remission. In follow-up studies, incidental abdominal tomography finding of an abdominopelvic lesion dependent on the mesentery at the level of the mesogastrium, smooth contours with fat-liquid level. Extension studies with negative tumor markers. Results. Due to high clinical and imaging suspicion of teratoma, the patient was taken to resection of the lesion. Histopathological examination confirmed the diagnosis of mature cystic teratoma of the mesentery. Conclusion. Mesenteric teratoma is a rare clinical entity and is considered one of the differential diagnoses of an abdominal mass with a compressive effect. Diagnosis is mainly based on clinical examination and imaging findings. Early surgical excision is the mainstay of treatment; laparoscopic or open approach depends on the clinical characteristics and the experience of the surgeon.

Influence of the addition of nanohydroxyapatite to scaffolds on proliferation and differentiation of human mesenchymal stem cells: a systematic review of in vitro studies

One of the main challenges of tissue engineering in dentistry is to replace bone and dental tissues with strategies or techniques that simulate physiological tissue repair conditions. This systematic review of in vitro studies aimed to evaluate the influence of the addition of nanohydroxyapatite (NHap) to scaffolds on cell proliferation and osteogenic and odontogenic differentiation of human mesenchymal stem cells. In vitro studies on human stem cells that proliferated and differentiated into odontogenic and osteogenic cells in scaffolds containing NHap were included in this study. Searches in PubMed/MEDLINE, Scopus, Web of Science, OpenGrey, ProQuest, and Cochrane Library electronic databases were performed. The total of 333 articles was found across all databases. After reading and analyzing titles and abstracts, 8 articles were selected for full reading and extraction of qualitative data. Results showed that despite the large variability in scaffold composition, NHap-containing scaffolds promoted high rates of cell proliferation, increased alkaline phosphatase (ALP) activity during short culture periods, and induced differentiation, as evidenced by the high expression of genes involved in osteogenesis and odontogenesis. However, further studies with greater standardization regarding NHap concentration, type of scaffolds, and evaluation period are needed to observe possible interference of these criteria in the action of NHap on the proliferation and differentiation of human stem cells.

Synergistic impact of -linolenic acid and -tocopherol on in vitro maturation and culture of buffalo oocytes

Abstract The objective of the current study was to investigate the synergistic impact of -Tocopherol and -Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50M -Tocopherol, GROUP 3: MM + 100 µM ALA + 100M -Tocopherol, GROUP 4: MM + 100 µM ALA + 200 M -Tocopherol and GROUP 5: MM + 100 µM ALA + 300 M -Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrodes Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of -Linolenic acid (100 µM) and -Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of -Linolenic acid (100 µM) in IVM media and increasing concentration of -tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of -tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of -tocopherol in maturation media having -Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Resumo O objetivo do presente estudo foi investigar o impacto sinérgico do -tocoferol e do ácido -linolênico (100 µM) na MIV e CIV de oócitos de búfala Nili Ravi. Os oócitos foram obtidos dos ovários de búfalos abatidos duas horas após o abate e levados ao laboratório. Complexos de oócitos cumulus de búfalo foram colocados aleatoriamente nos cinco grupos experimentais incluídos; GRUPO 1: Meio de maturação (MM) + 100 µM ALA (controle), GRUPO 2: MM + 100 µM ALA + 50 µM -tocoferol, GRUPO 3: MM + 100 µM ALA + 100 µM -tocoferol, GRUPO 4: MM + 100 µM ALA + 200 M -tocoferol e GRUPO 5: MM + 100 µM ALA + 300 M -tocoferol sob uma atmosfera de 5% de CO2 em ar a 38,5 °C por 22-24 h. A expansão cumulus e o estado de maturação nuclear foram determinados (Experimento 1). No experimento 2, os oócitos foram maturados como no experimento 1. Os oócitos maturados foram então fertilizados em meio de Tyrode's Albumina Lactato Piruvato (TALP) por cerca de 20 h e cultivados em meio de fluido oviductal sintético (SOF) para determinar o efeito do ácido -linolênico (100 µM) e -tocoferol em meio IVM em IVC de presumíveis zigotos. Para estudar o efeito do ácido -linolênico (100 µM) em meio IVM e aumentar a concentração de -tocoferol no meio de cultura no desenvolvimento inicial do embrião (Experimento 3), os presumíveis zigotos foram distribuídos aleatoriamente nos cinco grupos experimentais com concentração crescente de -tocoferol em meios de cultura. Maior porcentagem de oócitos em estágio MII no experimento 1 (65,2 ± 2,0), embriões em estágio de mórula no experimento 2 (30,4 ± 1,5) e experimento 3 (22,2 ± 2,0) foram obtidos. No entanto, os resultados gerais para a expansão das células do cumulus, maturação do oócito para o estágio MII e desenvolvimento embrionário subsequente entre os tratamentos permanecem estatisticamente semelhantes (P> 0,05). A suplementação de -tocoferol em meios de maturação com ácido -linolênico e / ou em meios de cultura de embriões não aumentou ainda mais a maturação in vitro de oócitos ou a produção de embriões.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

Influence of incubation temperature and embryonic motility on the growth of members of Caiman yacare (Daudin, 1802) / Influência da temperatura de incubação e da motilidade embrionária sobre o crescimento dos membros de Caiman yacare (Daudin, 1802)

Abstract This study aimed to evaluate whether skeletal development of the Pantanal Caiman (Caiman yacare) is similarly influenced by temperature variation and controlled increases in embryo motility. All eggs were incubated at 90% humidity and 29 °C for the first 45 days. Thereafter, the incubation temperature was either maintained at 29 °C and embryos were treated with 4-aminopyridine (4-AP) on days 46, 47, 48, and 49 (Group I, 29 °C 4-AP, n = 15); maintained at 29 °C (n = 14; Group II); or at 33 °C (n = 14, Group III). Embryonic movement was measured using an Egg Buddy® digital monitor on days 30, 35, 42, 49, 56, and 60, at which point embryos were euthanized and samples were collected for analysis. No differences were observed between groups with varying incubation temperatures. In contrast, embryonic motility was greater in embryos treated with 4-AP (P < 0.001) on day 49, and this was associated with higher proportions of snout-vent and hand lengths. This study demonstrates for the first time that pharmacologically induced increases in embryo motility result in phenotypic changes to the proportion of elements during prenatal ontogeny, thereby effectively altering the adaptation of the species to specific environments.

Resumo Este estudo objetivou avaliar os efeitos da temperatura e motilidade embrionária sobre o desenvolvimento esquelético de jacaré-do-pantanal (Caiman yacare). Os ovos foram incubados com 90% de umidade e empregou-se a temperatura de 29°C por 45 dias. Após, para a incubação do Grupo I a temperatura continuou em 29°C, mas associou-se à injeção de 4-aminopiridina (29°C-4AP, n = 15) aplicada nos dias 46, 47, 48 e 49, do Grupo II permaneceu em 29°C (n = 14) e do Grupo III elevou-se para 33°C (n = 14). A movimentação foi mensurada através do monitor digital Egg Buddy® nos dias 30, 35, 42, 49, 56 e 60 dias. Aos 60 dias, os embriões foram eutanasiados e coletadas amostras embrionárias. Na análise estatística não foram observadas diferenças entre os grupos para o fator temperatura sobre a motilidade embrionária no desenvolvimento esquelético. Em contraste, a motilidade evidenciou diferença estatística no dia 49 para o Grupo I (P < 0,001) e apresentou maiores proporções de nariz e mão. Esses dados demonstraram pela primeira vez que o aumento na motilidade, induzidos farmacologicamente resultam em divergências fenotípicas na proporção de segmentos anatômicos durante a ontogenia pré-natal, podendo alterar efetivamente a adaptação dos animais em ambientes específicos.

A comprehensive review on the documented characteristics of four Reticulitermes termites (Rhinotermitidae, Blattodea) of China / Uma revisão abrangente sobre as características documentadas de quatro cupins Reticulitermes da China

Termites are known as social insects worldwide. Presently in China 473 species, 44 genera and 4 families of termites have been reported. Of them, 111 Reticulitermes species are widely spread in different zones of China. The dispersion flight season of these Chinese Reticulitermes species are usually started from February to June, but in some regions different species are distributed, sharing their boundaries and having overlapping flight seasons. These reasons become important sources of hybridization between two different heterospecific populations of termites. It was confirmed that the fertilized eggs and unfertilized eggs of some Reticulitermes termites have the capacity of cleavage. While the unfertilized eggs of R. aculabialis, R. chinensis and R. labralis cleaved normally and the only R. aculabialis unfertilized eggs develop in embryos. While, the R. flaviceps and R. chinensis were observed with their abnormal embryonic development, and not hatching of eggs parthenogenetically. They were reported more threatening to Chinese resources as they propagate with parthenogenesis, hybridization and sexual reproduction. Eggshell and macrophiles of eggs play important roles in species identification and control. Although, they are severe pests and cause a wide range of damages to wooden structures and products in homes, buildings, building materials, trees, crops, and forests in China's Mainland.

Os cupins são conhecidos como insetos sociais em todo o mundo. Atualmente na China foram relatadas 473 espécies, 44 gêneros e 4 famílias de cupins. Destas, 111 espécies de Reticulitermes estão amplamente distribuídas em diferentes zonas da China. A temporada de voo de dispersão dessas espécies chinesas de Reticulitermes geralmente começa de fevereiro a junho, mas em algumas regiões diferentes espécies são distribuídas, compartilhando seus limites e tendo temporadas de voo sobrepostas. Essas razões tornam-se importantes fontes de hibridização entre duas populações heteroespecíficas de cupins. Foi confirmado que os ovos fertilizados e não fertilizados de alguns cupins Reticulitermes possuem capacidade de clivagem. Já os ovos não fertilizados de R. aculabialis, R. chinensis e R. labralis clivaram normalmente, e os únicos ovos não fertilizados de R. aculabialis se desenvolvem em embriões. R. flaviceps e R. chinensis foram observados com desenvolvimento embrionário anormal, e não eclosão de ovos por partenogênese. Eles foram relatados como mais ameaçadores para os recursos chineses à medida que se propagam com partenogênese, hibridização e reprodução sexual. Casca de ovo e macrófilos de ovos desempenham papéis importantes na identificação e controle de espécies, embora sejam pragas graves e causem uma ampla gama de danos a estruturas e produtos de madeira em residências, edifícios, materiais de construção, árvores, plantações e florestas na China continental.

Telocytes in Human Embryonic Development: A Preliminary Microscopic Anatomy Study / Telocitos en el Desarrollo Embrionario Humano: Un Estudio Preliminar de Anatomía Microscópica

SUMMARY: Telocytes are a cell population described in 2011 with a multitude of functions such as tissue support, regulation of stem cell niches or intercellular signal transmission. However, there are no studies about their embryonic origin, their function in development, or their moment of appearance. The objective of this work is to try to answer these questions through histological and immunofluorescence studies with samples from the embryological collection of the Department of Anatomy of the University of Granada. In the results obtained, as demonstrated by immunofluorescence for CD34, the presence of these cells can be seen in the fourth week of embryonic development in the perinotochordal region. Its presence is evident from the sixth week of development in a multitude of organs such as the heart, skeletal muscle tissue and supporting tissue of various organs such as the kidney, brain or pericardium. Its function seems to be when the embryonic histological images are analyzed in an evolutionary way, to act as a scaffold or scaffold for the subsequent population by mature tissue elements. In conclusion, telocytes appear at a very early stage of embryonic development and would have a fundamental role in it as scaffolding and directors of organ and tissue growth.

Los telocitos son una población celular descrita en 2011 con multitud de funciones como el sostén tisular, la regulación de los nichos de células madre o la transmisión de señales intercelulares. Sin embargo, no existen estudios acerca del origen embrionario de los mismos, su función en el desarrollo ni su momento de aparición. El objetivo de este trabajo es tratar de responder a estos interrogantes mediante estudios histológicos y por inmunofluorescencia con muestras de la colección embriológica del Departamento de Anatomía de la Universidad de Granada. En los resultados se puede observar como se demuestra mediante inmunofluorescencia para CD34, la presencia de estas células en la cuarta semana del desarrollo embrionario en la región perinotocordal. Su presencia se evidencia a partir de la sexta semana del desarrollo en multitud de órganos como corazón, tejidos músculo esqueléticos y tejidos de sostén de diversos órganos como riñón, encéfalo o pericardio. Su función parece ser al ser analizadas las imágenes histológicas embrionarias de forma evolutiva, la de actuar como un andamiaje o scafold para el posterior poblamiento por elementos tisulares maduros. Como conclusión, los telocitos aparecen en un momento muy precoz del desarrollo embrionario y presentarían una función fundamental en el mismo como andamiajes y directores del crecimiento de los órganos y tejidos.

Epidermoid cyst of the brain: A common cystic lesion in rare location

Epidermoid cysts are common benign tumors comprising around 1% and 2% of all intracranial tumors. Their usual locations include the parasellar region and cerebellopontine angle, and less commonly, the Sylvian fissure, suprasellar region, cerebral, and cerebellar hemispheres. Epidermoid cysts located in the brain stem are rare. These epidermoid cysts are similar to epidermoids arising in the skin which contain cheesy and flaky-white soft pultaceous material. Epidermoid cysts are very slow-growing tumors having a similar growth pattern of the epidermal cells of the skin and develop from the remnants of epidermal elements during the closure of the neural groove and disjunction of the surface ectoderm with neural ectoderm between the 3rd and 5th weeks of embryonic life. The ideal treatment of choice is the removal of cystic components with the complete resection of the capsule. We are presenting an interesting case of an epidermoid cyst in the frontal lobe in a 42-year-old male along with radiological investigations.

Study of ex-ovo embryonic development of Gallus gallus domesticus / Estudio inicial del desarrollo embrionario ex-ovo de Gallus gallus domesticus

SUMMARY: The domestic chicken is a species of bird that has been extensively studied in regard to its biology and as a model organism for science. The reproduction of the species is by the laying of fertilized eggs, which in a period of 21 days will develop a chick inside. Several methods have been described to develop embryos ex-ovo, allowing the observation and manipulation of the organism. This work has the propose to standardize a method that allows the development of the embryos inside the artificial incubation system, which has a low cost and is easy to make. In this work, 100 chicken eggs were used to study the effects of humidity, mineral supplementation, and the preincubation time of the egg on the incubation ex-ovo of the embryos. Embryo development was documented through the different days. Pulverized eggshell was selected as an optimal source to provide calcium, magnesium, phosphorus, and other minerals to the developing embryo. By providing 900-1200 mg of pulverized eggshell, 40 mL of the 0.001 % solution of benzalkonium chloride, and a preincubation time of approximately 56 h, the embryos were able to develop until 19 days, and even though they did not reach hatching, the incubation conditions that allowed the survival and development of embryos until late stages were achieved. Thus, due to the conditions established for calcium, humidity and preincubation time, in the present work, the chicks reached 19 days of development.

El pollo doméstico es una especie de ave que ha sido ampliamente estudiada en cuanto a su biología y como organismo modelo para la ciencia. La reproducción de la especie es por la puesta de huevos fecundados, que en un período de 21 días desarrollarán un polluelo en su interior. Se han descrito varios métodos para desarrollar embriones ex-ovo, permitiendo la observación y manipulación del organismo. Este trabajo tuvo como objetivo estandarizar un método que permita el desarrollo de los embriones dentro del sistema de incubación artificial, el cual tiene un bajo costo y es fácil de realizar. En este trabajo se utilizaron 100 huevos de gallina para estudiar los efectos de la humedad, la suplementación mineral y el tiempo de preincubación del huevo sobre la incubación ex-ovo de los embriones. El desarrollo embrionario se documentó a través de los diferentes días. Se seleccionó la cáscara de huevo pulverizada como una fuente óptima para proporcionar calcio, magnesio, fósforo y otros minerales al embrión en desarrollo. Al suministrar 900-1200 mg de cáscara de huevo pulverizada, 40 mL de la solución de cloruro de benzalconio al 0.001 % y un tiempo de preincubación de aproximadamente 56 h, los embriones lograron desarrollarse hasta los 19 días, y aunque no llegaron a eclosionar, los embriones lograron desarrollarse hasta los 19 días. Se lograron condiciones de incubación que permitieron la supervivencia y desarrollo de los embriones hasta etapas tardías. Así, debido a las condiciones establecidas de calcio, humedad y tiempo de preincubación, en el presente trabajo los pollitos alcanzaron los 19 días de desarrollo.

Distribution of calcitonin gene-related peptide and vascular endothelial growth factor in the mouse lung at the embryonic and postnatal stages / Distribución del péptido relacionado con el gen de la calcitonina y el factor de crecimiento endotelial vascular en el pulmón de ratón en las etapas embrionaria y posnatal

SUMMARY: Neuropeptide calcitonin gene-related peptide (CGRP) is a neurotransmitter related to vasculogenesis during organ development. The vascular endothelial growth factor A (VEGF-A) is also required for vascular patterning during lung morphogenesis. CGRP is primarily found in organs and initially appears in pulmonary neuroendocrine cells during the early embryonic stage of lung development. However, the relationship between CGRP and VEGF-A during lung formation remains unclear. This study investigates CGRP and VEGF-A mRNA expressions in the embryonic, pseudoglandular, canalicular, saccular, and alveolar stages of lung development from embryonic day 12.5 (E12.5) to postnatal day 5 (P5) through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. Further, we analyzed the expression of CGRP via immunohistochemistry. The VEGF-A mRNA was mainly scattered across the whole lung body from E12.5. CGRP was found to be expressed in a few epithelial cells of the canalicular and the respiratory bronchiole of the lung from E12.5 to P5. An antisense probe for CGRP mRNA was strongly detected in the lung from E14.5 to E17.5. Endogenous CGRP may regulate the development of the embryonic alveoli from E14.5 to E17.5 in a temporal manner.

El péptido relacionado con el gen de la calcitonina (CGRP) es un neurotransmisor vinculado con la vasculogénesis durante el desarrollo de órganos. El factor de crecimiento endotelial vascular A (VEGF-A) también se requiere para el patrón vascular durante la morfogénesis pulmonar. El CGRP se encuentra principalmente en los órganos y aparece inicialmente en las células neuroendocrinas pulmonares durante la etapa embrionaria temprana del desarrollo pulmonar. Sin embargo, la relación entre CGRP y VEGF-A durante la formación de los pulmones sigue sin estar clara. Este estudio investiga las expresiones de ARNm de CGRP y VEGF-A en las etapas embrionaria, pseudoglandular, canalicular, sacular y alveolar del desarrollo pulmonar desde el día embrionario 12,5 (E12,5) hasta el día postnatal 5 (P5) a través de la reacción en cadena de la polimerasa cuantitativa en tiempo real. (qRT-PCR) e hibridación in situ. Además, analizamos la expresión de CGRP mediante inmunohistoquímica. El ARNm de VEGF-A se dispersó principalmente por todo parénquima pulmonar desde E12,5. Se encontró que CGRP se expresaba en unas pocas células epiteliales de los bronquiolos canaliculares y respiratorios del pulmón desde E12,5 a P5. Se detectó fuertemente una sonda antisentido para ARNm de CGRP en el pulmón de E14,5 a E17,5. El CGRP endógeno puede regular el desarrollo de los alvéolos embrionarios de E14,5 a E17,5 de manera temporal.

Presentación atípica de teratoma quístico maduro gigante en una paciente adolescente: a propósito de un caso / Atypical presentation of a giant mature cystic teratoma in an adolescent patient: A case report

Introducción: El teratoma quístico maduro es un tipo de tumor derivado de las células germinales que aparece en pacientes en edad fértil. La edad más frecuente de aparición de este tipo de tumores es entre los 20 y40 años.Caso clínico: Se presenta el caso de una paciente adolescente de 18 años con masa abdominal gigante de crecimiento abrupto cuya presentación fue atípica dado el tamaño de esta, el cual se manifestó con dolor abdominal agudo.Tratamiento: Se realiza resección de la masa la cual confirma el diagnóstico histopatológico de teratoma quístico maduro.Conclusión: Este tipo de patologías rara vez se presentan con un crecimiento tan exagerado como el caso de la paciente en mención, y la resolución quirúrgica sigue siendo el gold estándar en cuanto al tratamiento.Palabras clave:DeCS: Teratoma, Células germinativas embrionarias, Adolescente, Neoplasias

Introduction: Mature cystic teratoma is a type of tumor derived from germ cells that appears in patients of childbearing age. The most common age at which this type of tumor appearsis 20 to40.Clinical case: The case of an 18-year-old adolescent patient with a giant abdominal mass of abrupt growth is presented, whose presentation was atypical given its size, which manifested with acute abdominal pain Treatment: Amass resection confirmedthehistopathological diagnosis of mature cystic ter-atoma.Conclusion: This type of pathology rarely presents with growth as exaggerated as in the case of the patient mentioned. Surgicalresolution continues to be the gold standard in terms of treatment

Effect of Qigongwan on Quality of Oocytes and Embryonic Development Potential in Patients with PCOS Infertility with Phlegm-dampness Syndrome / 中国实验方剂学杂志

ObjectiveTo observe the regulation of Qigongwan on the expression of proliferation and apoptosis-related factors programmed cell death 4 （PDCD4） and proliferating cell nuclear antigen （PCNA） in ovarian granulosa cells （GCs） in patients with polycystie ovarian syndrome （PCOS） infertility with phlegm-dampness syndrome， and to explore the effect of Qigongwan on the quality of oocytes and embryonic development potential. MethodSixty-six patients with PCOS with phlegm-dampness syndrome who underwent in vitro fertilization and embryo transfer （IVF-ET） were randomly selected and divided into an observation group （Qigongwan + western medicine） and a control group （western medicine）， with 33 patients in each group. Antagonist regimen was used to promote ovulation in the two groups. The observation group was given Qigongwan one cycle before IVF based on the treatment of conventional western medicine， while the control group was not given Chinese medicine. The improvement of phlegm and dampness syndrome， the dosage and the number of days of using gonadotropins （Gn）， the levels of luteinizing hormone （LH）， estradiol （E2）， and progesterone （P） on the day of human chorionic gonadotropin（HCG） injection， the 2PN fertilization rate， and the high-quality embryo rate of patients in the two groups were compared. Real-time polymerase chain reaction （Real-time PCR） and Western blot technology were used to detect the expression of PCNA and PDCD4 in GCs. ResultAs compared with groups before treatment， the score of phlegm-dampness syndrome in both groups was significantly lower （P<0.01）. The score of phlegm and dampness syndrome in the observation group was significantly lower than that of the control group （P<0.01）. As compared with the control group， the levels of LH， E2， and P in the observation group was higher， but only the difference in the level of E2 was statistically significant （P<0.01）. The 2PN fertilization rate ［82.25% （556/676） vs 69.92% （365/522）， χ2=25.172， P<0.01］ and high-quality embryo rate ［44.19% （190/430） vs 34.23% （102/298）， χ2=7.266， P<0.01］ in the observation group were significantly higher than that of the control group （P<0.01）. As compared with the control group， the mRNA and protein expression of PDCD4 in ovarian GCs was down-regulated in the observation group and that of PCNA was up-regulated （P<0.05）. ConclusionBy down-regulating the expression of PDCD4 and up-regulating the expression of PCNA， Qigongwan may interfere with follicle development， adjust hormone levels， improve the symptomatic manifestations of patients with PCOS with phlegm-dampness syndrome， inhibit the apoptosis of GCs， and promote growth， thus improving the quality of oocytes and embryonic development potential.

Non-targeted LC-MS profiling reveals association between human early embryo grades and levels of amino acids and carnitines in embryo culture medium / 中华检验医学杂志

Objective:In order to determine the development potential of human embryos in vitro, amino acid and carnitine levels were measured in the culture medium of different grades of early human embryos. Methods:From the infertile couples who received in vitro fertilization-embryo transfer treatment in the Department of Reproductive Medicine of Linyi People′s Hospital from June 2022 to December 2022, the age of the women was defined as 25-35 years old [31.5(26.5, 33.25)] with 8-20 eggs, 126 cultured cells and embryos of the third day were randomly collected from infertile couples. They were divided into three groups according to the morphological level of the corresponding embryos: excellent, neutral and poor. Amino acids and L-carnitines levels in culture medium were detected by liquid chromatography-mass spectrometry (LC-MS). Using analysis of variance to compare differences among groups, correlation analysis, factor analysis was performed to analyze the association between the levels of amino acids and L-carnitines and development potential of early human embryos.Results:The value of Methionine/Phenylalanine was found statistically different among superior embryo (3.09±1.67), moderate embryo (4.00±1.19) and inferior embryo (4.99±2.04). The difference between the three groups was statistically different ( F=7.09, P<0.05): superior embryo vs moderate embryo ( t=-0.91, P<0.05), superior embryo vs inferior embryo ( t=-1.91, P<0.05), moderate embryo vs inferior embryo ( t=-0.99, P<0.05). Among different amino acids, Phe had the strongest positive correlation with Tyr ( r=0.99, P<0.01). Among different carnitines, C 8/C 10 has the strongest positive correlation with C 5DC+C 6OH/C 16( r=0.44, P<0.01). The weight value of leucine (isoleucine), arginine, valine/phenylalanine, glycine, tyrosine and carnitine(C 5DC+C 6OH)/C 8 calculated by the least square fitting model is 2.22, 1.99, 1.65, 1.54, 1.21 and 1.15 respectively. Conclusion:Leucine, arginine, valine/phenylalanine, glycine, tyrosine and carnitine (C 5DC+C 6OH)/C 8 in embryo culture medium were significantly correlated with the levels of early human embryos in vitro.

Role of ROS/TXNIP/NLRP3 signaling pathway in high glucose-induced pyroptosis of human embryonic trophoblast cells / 中华内分泌外科杂志

Objective:To investigate the changes of ROS/TXNIP/NLRP3 signaling pathway in pyroptosis of human embryonic trophoblast cells induced by high glucose.Methods:Human embryonic trophoblast cells were cultured in vitro to establish high glucose injury model, and they were randomly divided into control group, high glucose (HG) group and HG + ROS inhibitor N-acetyl-L-cysteine (HG + NAC) group. MTT assay was used to detect the cell survival rate. The level of ROS in each group was detected by dihydroethidine ROS fluorescence probe. Expression of TXNIP and NLRP3 mRNA was detected by real-time quantitative PCR (RT-qPCR). Western blot analysis was used to detect the expression levels of TXNIP, NLRP3, Caspase-1, interleukin (IL) -1β, tumor necrosis factor-α (TNF-α) and GSDMD proteins. In addition, pyroptosis was detected by flow cytometry.Results:The optimal glucose concentration for high glucose-induced injury of human embryonic trophoblast cells was 30 mmol/L. Compared with the control group (96.27±3.10) %, the survival rate of human embryonic trophoblast cells in HG group (55.44±2.15) % was significantly lower ( P<0.05), while the fluorescence intensity (ROS level) of 7 'dichlorofluorescein (DCF), the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly higher ( P<0.05) ; Compared with HG group, the survival rate of human embryonic trophoblast cells in HG+NAC group (84.75±2.33) % was significantly higher ( P<0.05), the fluorescence intensity (ROS level) of DCF, the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, and expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly lower ( P<0.05) . Conclusion:Inhibition of ROS level in human embryonic trophoblast cells induced by high glucose may promote cell proliferation and reduce the occurrence of pyroptosis by inhibiting TXNIP/NLRP3 signaling pathway.

Effects of manipulating lactate dehydrogenase gene on metabolism of HEK-293 and production of human adenovirus / 生物工程学报

Reducing lactate accumulation has always been a goal of the mammalian cell biotechnology industry. When animal cells are cultured in vitro, the accumulation of lactate is mainly the combined result of two metabolic pathways. On one hand, glucose generates lactate under the function of lactate dehydrogenase A (LDHA); on the other hand, lactate can be oxidized to pyruvate by LDHB or LDHC and re-enter the TCA cycle. This study comprehensively evaluated the effects of LDH manipulation on the growth, metabolism and human adenovirus (HAdV) production of human embryonic kidney 293 (HEK-293) cells, providing a theoretical basis for engineering the lactate metabolism in mammalian cells. By knocking out ldha gene and overexpression of ldhb and ldhc genes, the metabolic efficiency of HEK-293 cells was effectively improved, and HAdV production was significantly increased. Compared with the control cell, LDH manipulation promoted cell growth, reduced the accumulation of lactate and ammonia, significantly enhanced the efficiency of substrate and energy metabolism of cells, and significantly increased the HAdV production capacity of HEK-293 cells. Among these LDH manipulation measures, ldhc gene overexpression performed the best, with the maximum cell density increased by about 38.7%. The yield of lactate to glucose and ammonia to glutamine decreased by 33.8% and 63.3%, respectively; and HAdV titer increased by at least 16 times. In addition, the ATP production rate, ATP/O2 ratio, ATP/ADP ratio and NADH content of the modified cell lines were increased to varying degrees, and the energy metabolic efficiency was significantly improved.

CXCR5 Regulates Neuronal Polarity Development and Migration in the Embryonic Stage via F-Actin Homeostasis and Results in Epilepsy-Related Behavior / 神经科学通报·英文版

Epilepsy is a common, chronic neurological disorder that has been associated with impaired neurodevelopment and immunity. The chemokine receptor CXCR5 is involved in seizures via an unknown mechanism. Here, we first determined the expression pattern and distribution of the CXCR5 gene in the mouse brain during different stages of development and the brain tissue of patients with epilepsy. Subsequently, we found that the knockdown of CXCR5 increased the susceptibility of mice to pentylenetetrazol- and kainic acid-induced seizures, whereas CXCR5 overexpression had the opposite effect. CXCR5 knockdown in mouse embryos via viral vector electrotransfer negatively influenced the motility and multipolar-to-bipolar transition of migratory neurons. Using a human-derived induced an in vitro multipotential stem cell neurodevelopmental model, we determined that CXCR5 regulates neuronal migration and polarization by stabilizing the actin cytoskeleton during various stages of neurodevelopment. Electrophysiological experiments demonstrated that the knockdown of CXCR5 induced neuronal hyperexcitability, resulting in an increased number of seizures. Finally, our results suggested that CXCR5 deficiency triggers seizure-related electrical activity through a previously unknown mechanism, namely, the disruption of neuronal polarity.

Preliminary experimental study on the prevention of all-trans retinoic acid induced cleft palate in mice by nicotinamide / 口腔疾病防治

Objective@# To explore the preventive effect of nicotinamide (NAM) on cleft palate induced by all-trans retinoic acid (RA), to provide research evidence for the prevention of cleft palate. @*Methods @#The mouse cleft palate model was induced by intragastric administration of 70 mg/kg all-trans retinoic acid at embryonic day 10.5 (E10.5) in the control group. The mouse cleft palate model was treated by caudal vein injection of 20 mg/kg NAM at E8.5 to E13.5 in the experimental group (1). The cleft palate model was treated by caudal vein injection of 40 mg/kg NAM at E8.5-E13.5 in the experimental group (2). The cleft palate of fetal rats was observed by laparotomy on E16.5 and statistically analyzed. Annexin V-FITC/PI double staining was used to detect the apoptosis of mouse embryonic palatal mesenchyme (MEPM) cells treated with RA 1 μmol/L (RA 1 group), NAM 200 μmol/L (NAM 200 group), and both NAM 200 μmol/L and RA 1 μmol/L (NAM 200+RA 1 group) for 24 hours by flow cytometry and the apoptosis rate in groups were compared. Culture without RA or NAM was used as a control. @*Results @# The cleft palate rate in the control group was 98%. The cleft palate rate in experimental group (1) was 87%. There was no significant difference between groups (P>0.05). The cleft palate rate in the experimental group (2) was 63%, compared with the control group, there was a significant difference (P<0.01). The cell apoptosis rate was 16.53%±2.89% in the CONTROL group. The cell apoptosis rate was 22.9%±1.85% in the RA 1 group, which was a significant increase compared with the CONTROL group (P<0.01). The apoptotic rate of the NAM 200 group was 9.23%±1.39%, which was a significant decrease compared with NA 1 group (P<0.01). The apoptosis rate of the NAM 200+RA 1 group was 14.9%±7.67%, which was a significant decrease compared with the RA 1 group (P<0.01).@*Conclusion@#NAM can prevent cleft palate. 40 mg/kg nicotinamide during pregnancy is an effective concentration for the prevention of RA-induced cleft palate. The mechanism by which NAM prevents cleft palate may be that NAM inhibits RA-induced apoptosis of MEPM cells.

Mild overexpression of a novel transcription factor Grainy head like-2 (Grhl2) elicits hybrid phenotype in human astrocytoma cells

The Grainy head like-2 (Grhl2) transcription factor plays a major role in embryonic and cancer development. The role of Grhl2 has been intensively studied in various cancers but not for brain cancer. Hence, in this study, we provide a preliminary understanding on the role of Grhl2 that regulate the transition of astrocytoma cells. The human A172 astrocytoma cell line, a mesenchymal cell characterized by mild overexpression of Grhl2 transcription factor, was used in this study. At first, the Grhl2 stably overexpressing A172 clones into three types i.e., Grhl2+ (mild), Grhl2++ (moderate) and Grhl2+++ (high) based on mRNA and protein expression levels of Grhl2 were characterized. Phenotypic characteristics of vector and Grhl2+ cells were observed using phase contrast microscopy. Quantitative PCR (qPCR), Western blot and immunofluorescence were used to detect the level of mesenchymal markers (N-cadherin/vimentin) and also epithelial markers (E-cadherin/ ?- catenin) in vector and Grhl2+ cells. The migration and invasion characteristics of vector and Grhl2+ cells were determined by scratch assay and Boyden chamber assay. Further, the Grhl2+ cells were characterized to determine the effect of temozolomide chemotherapy drug which were widely used in treating brain cancer. As expected, in phase contrast image, we observed the mesenchymal characteristic of A172 cells becomes hybrid phenotype i.e., mixture of mesenchymal (spindle-like fibroblast morphology) and epithelial (cobblestone like appearance) cells upon Grhl2 mild expression (Grhl2+) when compared to vector cells. Further, we found that there was a significant upregulation of E-cadherin at both mRNA and protein levels in Grhl2+ cells when compared to vector cells. There was a significant upregulation of ?-catenin, N-cadherin and vimentin at mRNA levels, but there was no significant upregulation at the protein levels in Grhl2+ cells compared to the vector cells. The migration and invasion were diminished in Grhl2+ cells when compared to the vector control cells. We observed that the Grhl2+ were sensitive to the temozolomide compared to the vector cells. This infers that the Grhl2+ cells were unable to attain complete transition of mesenchymal to epithelial state, and hence we categorized the Grhl2+ cells as hybrid phenotype. The results provide a better understanding of the largely unknown function of Grhl2 in human astrocytoma cells as tumor progression or suppression.

Asynchrony between in vivo and in vitro rabbit embryos

Background: Comparative features of embryos developed under in vitro and in vivo conditions are particularly important in designing embryo transfer procedures that fulfil embryo-recipient synchronization requirements. Objective: To determine the degree of asynchrony in rabbit embryo development between cultured and in vivo embryos. Methods: A total of 55 non- lactating multiparous female rabbits were used. Embryos were classified as 16-cells or early morulae at 48 hours post-coitum (hpc). Embryos were cultured during 30 or 32 h and embryo development was compared with in vivo embryos of 72 hpc. In vitro and in vivo embryos at 72 hpc were classified as early or compacted morulae. Bayesian statistics was used. Difference between in vivo and in vitro embryos and the actual probability of the difference between the in vivo and in vitro embryo higher than zero (P) was estimated. Results: The percentage of compacted morulae was higher in in vivo embryos than in in vitro embryos with +6 h of asynchrony (73.5 and 32.8%, P=1.00). But the percentage of compacted morulae was similar with +8 h asynchrony. Conclusions: In vitro embryos delay their development by + 8 hours compared to in vivo embryos.

Antecedentes: El desarrollo comparativo de embriones producidos in vitro e in vivo es particularmente importante para el diseño de procedimientos de transferencia de embriones cuando se requiere sincronización entre el embrión y la hembra receptora. Objetivo: Determinar el grado de asincronía en el desarrollo embrionario entre embriones in vivo y cultivados. Métodos: Un total de 55 conejas multiparas no lactantes fueron utilizadas. Los embriones se clasificaron en 16 células o mórulas tempranas a las 48 horas después del coito (hpc). Los embriones se cultivaron durante 30 ó 32 horas y el desarrollo embrionario se comparó con embriones de 72 hpc obtenidos in vivo. Los embriones in vitro e in vivo a 72 hpc se clasificaron como mórulas tempranas o compactas. Se utilizó estadística bayesiana. Se estimó la diferencia entre embriones in vivo e in vitro y la probabilidad de que la diferencia sea superior a cero (P). Resultados: El porcentaje de mórulas compactas fue mayor en embriones in vivo que en embriones in vitro con +6 horas de asincronía (73,5 y 32,8%, P=1,00), pero el porcentaje de mórulas compactas fue similar con asincronía de +8 horas. Conclusión: Los embriones cultivados retrasan +8 horas su desarrollo en comparación con los embriones in vivo.

Antecedentes: A aquisição do desenvolvimento de embriões produzidos in vitro e in vivo é particularmente importante na concepção de procedimentos de transferência de embriões em que a sincronização entre o embrião e a fêmea receptora é necessária. Objetivo: Determinar o grau de assincronia no desenvolvimento embrionário entre embriões cultivados e in vivo. Métodos: Um total de 55 coelhos multíparos não lactantes foram usados. Os embriões foram classificados em 16 células ou mórulas iniciais 48 horas de gestação (hpc). Os embriões foram cultivados por 30 ou 32 horas e o desenvolvimento embrionário foi comparado com embriões de 72 hpc obtidos in vivo. Embriões in vitro e in vivo a 72 hpc foram classificados como mórulas precoces ou compactadas. Estatísticas bayesianas foram usadas. A diferença entre embriões in vivo e in vitro e a probabilidade de que a diferença seja maior que zero (P) foi estimada. Resultados: A porcentagem de mórulas compactadas foi maior em embriões in vivo do que em embriões in vitro com +6 horas de assincronia (73,5 e 32,8%, P=1,00). Mas a porcentagem de mórulas compactadas foi semelhante com assincronia de +8 horas. Conclusão: Embriões cultivados atrasam seu desenvolvimento em +8 horas em comparação com embriões in vivo.