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Morphological Alterations in Human Skeletal Muscle Cells Exposed to di-(2-Ethylhexyl) Phthalate (dehp) in Primary Cell Culture Conditions

Alteraciones Morfológicas en Células Musculares Esqueléticas Humanas Expuestas a Ftalato de di-(2-etilhexilo) (dehp) en Condiciones de Cultivo Celular Primario

Pablo Hernández Almaraz ORLANDO LUGO LUGO Ramón Gaxiola Robles OSCAR KURT BITZER QUINTERO Luis Javier Ramírez Jirano Elizabeth Brassea Pérez TANIA ZENTENO SAVIN (2022, [Artículo])

"Di-(2-ethylhexyl) phthalate (DEHP) is among the most common plasticizer additives that humans are in contact with daily. DEHP can be released from plastic and enter the human body, whereby it is metabolized and transformed into oxidative hydrophilic molecules. Clinical follow-ups in patients exposed to this phthalate and investigations in cultures of several cell types have provided information on its effects. For example, it is associated with inhibition of diploid human cell development and morphological changes in cultured germ cells. Although skeletal muscle represents around 50 % of the human body mass, knowledge about the effects of DEHP on this tissue is poor. Cultured skeletal muscle cells were exposed to DEHP (1 mM) for 13 days with the aim of exploring and evaluating some of the potential morphological effects. Three culture development parameters and nine cell characteristics were monitored during the bioassay. At 13 days, growth area, cell viability, and concentration of total proteins were lower in DEHP exposed than in control cells. Cell width and area, as well as the diameter of the nucleus and nucleolus, were greater in exposed cells than in control cells. These are interpreted as signs of cytotoxicity and suggest potential adverse effects on the development of skeletal muscle cells from DEHP exposure, as reported for other cell types."

Emerging pollutants, Morphometry, Phthalates, Plasticizers, Toxicity MEDICINA Y CIENCIAS DE LA SALUD CIENCIAS MÉDICAS BIOLOGÍA HUMANA FISIOLOGÍA DEL MÚSCULO FISIOLOGÍA DEL MÚSCULO

EVALUACIÓN DE LA ACTIVIDAD Y EXPRESIÓN GÉNICA DE ENZIMAS ANTIOXIDANTES (SOD, GPX, GST, GR y CAT) COMO BIOMARCADORES DE EXPOSICIÓN A FTALATOS EN CULTIVO CELULAR

Elizabeth Brassea Pérez (2023, [Tesis de doctorado])

"El plastificante bis(2-etilhexilo) ftalato (DEHP) altera el equilibrio entre la producción de especies reactivas de oxígeno y las defensas antioxidantes. A pesar de la información disponible sobre los efectos tóxicos del DEHP en organismos modelo, la respuesta celular al DEHP en especies naturalmente adaptadas para tolerar el estrés oxidativo aún no se ha explorado. Los mamíferos marinos están naturalmente adaptados para hacer frente al estrés oxidativo derivado de la isquemia/reperfusión inducida por el buceo. El objetivo de este estudio fue comparar los cambios en los indicadores de estrés oxidativo asociados al DEHP en células de músculo esquelético de humanos (Homo sapiens sapiens) y lobos marinos de California (Zalophus californianus). Se recolectaron muestras de músculo abdominal de mujeres sanas sometidas a cesárea programada, tras obtener su consentimiento informado. Las muestras de músculo de lobo marino se obtuvieron de crías encontradas recientemente muertas en la colonia de Los Islotes, Golfo de California. Las células de músculo esquelético se aislaron y cultivaron en condiciones estándar. Las células se dividieron en dos grupos. Un grupo fue expuesto a DEHP (1 mM) durante 13 días (n=25; exposición), el otro grupo se mantuvo bajo condiciones control, es decir sin DEHP añadido (n=25; control). Se midió la producción de radical superóxido (O2•-), el daño oxidativo, la actividad de enzimas antioxidantes y la expresión génica mediante métodos espectrofotométricos y RT-qPCR. En células expuestas al DEHP la producción de O2•- y la actividad de superóxido dismutasa (SOD) fueron mayores que en las células bajo condiciones control para ambas especies. La actividad de glutatión S-transferasa (GST) y los niveles de carbonilos proteicos (PC) aumentaron significativamente en las células humanas expuestas a DEHP, y no significativamente en las células de lobo marino. Por el contrario, las actividades de glutatión peroxidasa (GPx) y catalasa (CAT) aumentaron significativamente en las células de lobo marino, pero no en las de humano expuestas a DEHP. Se observó una expresión diferencial de los genes implicados en el metabolismo redox entre las células control y expuestas a DEHP y entre especies. Los patrones de expresión de 26 y 19 genes fueron significativamente diferentes en las células humanas y de lobo marino, respectivamente, después de la exposición a DEHP. En las células humanas, la expresión de la GST1 microsomal y la GST (κ, μ, θ, ω y ᴢ) fue mayor,

"The plasticizer bis (2-ethylhexyl) phthalate (DEHP) dysregulates the balance between reactive oxygen species production and antioxidant defenses. Despite the information available on DEHP’s hazardous effects in model species, the cellular response to DEHP in organisms naturally adapted to tolerate oxidative stress remains unexplored. Marine mammals are naturally adapted to cope with oxidative stress derived from diving-induced ischemia/reperfusion. The objective of this study was to compare changes in oxidative stress indicators induced by DEHP in human (Homo sapiens sapiens) and California sea lion (Zalophus californianus) skeletal muscle cells. Abdominal muscle samples were collected from healthy women undergoing planned cesarean surgery at full term after obtaining informed consent. Sea lion muscle samples were obtained from recently deceased pups at Los Islotes rookery, Gulf of California. Skeletal muscle cells were isolated and cultured under standard conditions. Cells were divided into two groups. One group was exposed to 1 mM DEHP for 13 days (n=25; treatment), and the other served as a control (n=25; no DEHP). Superoxide radical (O2•-) production, oxidative damage, antioxidant enzyme activities and gene expression were measured using spectrophotometric methods and RT-qPCR. DEHP exposure increased O2•- production and superoxide dismutase (SOD) activity in both species. While the activity of glutathione S-transferase (GST) and protein carbonyls (PC) levels significantly increased in human cells, these indicators showed a non-significant elevation in sea lion cells. In contrast, glutathione peroxidase (GPx) and catalase (CAT) activities increased significantly in sea lion but not in human cultures exposed to DEHP. Genes involved in redox metabolism showed differential expression between treatments and species. DEHP exposure led to widespread modifications in gene expression patterns, with 26 and 19 differentially expressed genes in human and sea lion cells, respectively. In human cells, DEHP increased GST1 and GST (κ, μ, θ, ω, and ᴢ), while suppressing 8-oxoguanine DNA glycosylase (OGG1), CAT, GR, and nuclear factor erythroid 2-related factor 2 (NRF2) expression, suggesting increased oxidative stress and phase two detoxification processes. In contrast, DEHP increased OGG1, NRF2, GPx2 and SOD3 expression, suggesting that DEHP activates antioxidant defenses in sea lion cells, potentially contributing to maintain redox homeostasis and avoid oxidative...

Contaminantes emergentes, enzimas antioxidantes, estrés oxidativo, mamíferos, Una Salud Antioxidant enzymes, emerging pollutants, mammals, One Health, oxidative stress BIOLOGÍA Y QUÍMICA QUÍMICA BIOQUÍMICA ENZIMOLOGIA ENZIMOLOGIA

Validación del potencial antioxidante de citrus aurantium en Tabasco, México

Validation of the antioxidant potential of citrus aurantium in Tabasco, Mexico

CLAUDINA VILLARREAL MARIA TERESA CADENAS GONZALEZ Francisca Méndez Morales GLORIA IVETTE BOLIO LÓPEZ Manuel Mateo Hernández Villegas Nidia Elena Rivera DAMIANYS ALMENARES LOPEZ Catalina Rivas Morales (2023, [Artículo])

Citrus aurantium L. (Rutaceae), comúnmente conocida como naranja amarga, posee múltiples potenciales terapéuticos. Se realizó un estudio cualitativo experimental con el objetivo de conocer las características del consumo de C. aurantium (CA) en población tabasqueña. Para validar su uso, se preparó un extracto etanólico (85 %) de las hojas secas de CA y se realizó el tamizaje fitoquímico, con posterior cuantificación del contenido de fenoles y flavonoides totales. Posteriormente se determinó actividad antioxidante por el ensayo DPPH y toxicidad aguda con Artemia Salina. Las partes más empleadas de la planta son las hojas para aliviar dolores musculares, síntomas de resfriados. En el estudio fitoquímico se encontraron que el extracto contiene alcaloides, flavonoides, cumarinas, quinonas y taninos. En la cromatografía de capa fina del extracto hidroalcohólico se demostró la presencia de la naringina. Se obtuvo un contenido de fenoles totales de 69.42 ± 3.47 EAG/g MS y de flavonoides totales de 14.78 ± 0.28 EQ/g MS. Contiene actividad antioxidante de 9240 ± nmol TEAC/gMS y el ensayo de toxicidad aguda demostró una toxicidad moderada. Los resultados evidencian que el extracto etanólico al 85 % contiene compuestos fenólicos y flavonoides, permitiendo sostener las propiedades medicinales y farmacológicas conferidas a la planta en el estado de Tabasco.

Citrus aurantium L. (Rutaceae), commonly known as bitter orange, has multiple therapeutic potentials. An experimental qualitative study was carried out with the objective of knowing the characteristics of the consumption of C. aurantium (CA) in the Tabasco population. To validate its use, an ethanolic extract (85 %) was prepared from the dry CA leaves and phytochemical screening was performed, with subsequent quantification of the content of total phenols and flavonoids. Subsequently, antioxidant activity was determined by the DPPH assay and acute toxicity with Artemia salina. The most used parts of the plant are the leaves to relieve muscle pain, cold symptoms. In the phytochemical study it was found that the extract contains alkaloids, flavonoids, coumarins, quinones and tannins. In the thin layer chromatography of the hydroalcoholic extract, the presence of naringin was demonstrated. A total phenol content of 69.42 ± 3.47 EAG/g DM and total flavonoids of 14.78 ± 0.28 EQ/g DM were obtained. It contains antioxidant activity of 9240 ± nmol TEAC/gMS and the acute toxicity test showed moderate toxicity. The results show that the 85% ethanolic extract contains phenolic compounds and flavonoids, allowing to sustain the medicinal and pharmacological properties conferred to the plant in the state of Tabasco.Conclusions: this study showed the existence of phenols and flavonoids, the antioxidant activity of hydroalcoholic extracts from C. aurantium leaves and their moderate toxicity against Artemia salina.

Conflicto de interés: Los autores declaran que no existe ningún conflicto de intereses.

Contribución por autor: DAL: diseño, escritura y revisión del documento, MTCG, MMHV, GIBL, NERL, FMM, ECVI, CRM: asesoría, escritura y revisión del documento.

Financiación o fondos: El presente estudio no conto con ningún apoyo financiero, todo fue con recurso propio de los investigadores.

Conflicto de interés: Los autores declaran que no existe ningún conflicto de intereses.

Contribución por autor: DAL: diseño, escritura y revisión del documento, MTCG, MMHV, GIBL, NERL, FMM, ECVI, CRM: asesoría, escritura y revisión del documento.

Financiación o fondos: El presente estudio no conto con ningún apoyo financiero, todo fue con recurso propio de los investigadores.

Citrus aurantium Antioxidante Fenoles Flavonoides Toxicidad aguda Citrus aurantium Antioxidant Phenol Flavonoids Acute toxicity INGENIERÍA Y TECNOLOGÍA CIENCIAS TECNOLÓGICAS OTRAS ESPECIALIDADES TECNOLÓGICAS OTRAS OTRAS

Chronic toxicity of shrimp feed added with silver nanoparticles (Argovit-4®) in Litopenaeus vannamei and immune response to white spot syndrome virus infection

Carlos Rosendo Romo Quiñonez Píndaro Álvarez Ruiz CLAUDIO HUMBERTO MEJIA RUIZ NINA BOGDANCHIKOVA Alexey Pestryakov CARINA GAMEZ JIMENEZ WENCESLAO VALENZUELA QUIÑONEZ Magnolia Montoya Mejía EUSEBIO NAVA PEREZ (2022, [Artículo])

"In recent years, the application of silver nanoparticles (AgNPs) as antibacterial compounds has been widely used in human and veterinary medicine. In this work, we investigated the effects of AgNPs (Argovit-4 R ) as feed additives (feed-AgNPs) on shrimp (Litopenaeus vannamei) using three different methods: 1) chronic toxicity after 28 days of feeding, 2) Effects against white spot syndrome virus (WSSV) challenged by oral route, and 3) transcriptional responses of immune-related genes (PAP, ProPO, CTL-3, Crustin, PEN3, and PEN4) following WSSV infection. The results showed that the feed-AgNPs did not interfere with the growth and survival of shrimp. Also, mild lesions in the hepatopancreas were recorded, proportional to the frequency of the feed-AgNP supply. Challenge test versus WSSV showed that feeding every 7 days with feed-AgNPs reduced mortality, reaching a survival rate of 53%, compared to the survival rates observed in groups fed every 4 days, daily and control groups of feed-AgNPs for the 30%, 10%, and 7% groups, respectively. Feed-AgNPs negatively regulated the expression of PAP, ProPO, and Crustin genes after 28 days of treatment and altered the transcriptional responses of PAP, ProPO, CTL-3, and Crustin after WSSV exposure. The results showed that weekly feeding-AgNPs could partially prevent WSSV infection in shrimp culture. However, whether or not transcriptional responses against pathogens are advantageous remains to be elucidated."

Silver nanoparticles, Shrimp, Aquaculture, Chronic toxicity, WSSV, AgNP, Argovit, Litopenaeus vannamei , Silver fed, White spot syndrome virus BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA INMUNOLOGÍA INMUNIZACIÓN INMUNIZACIÓN