A full understanding of the molecular mechanism of action of choline kinase α (ChoKα) inhibitors at the cell level is essential for developing therapeutic and preventive approaches for cancer. The ...aim of the present study was to evaluate the effects of the ChoKα inhibitors EB-3D and EB-3P on lipid metabolism in HepG2 cells. We used methyl-
Ccholine, 1,2-
Cacetic acid and 2-
Hglycerol as exogenous precursors of the corresponding phospholipids and neutral lipids. Methyl-
Ccholine was also used to determine choline uptake. Protein levels were determined by Western blot. Ultrastructural alterations were investigated by transmission electron microscopy. In this work, we demonstrate that EB-3D and EB-3P interfere with phosphatidylcholine biosynthesis via both CDP-choline pathway and choline uptake by the cell. Moreover, the synthesis of both diacylglycerols and triacylglycerols was affected by cell exposure to both inhibitors. These effects were accompanied by a substantial decrease in cholesterol biosynthesis, as well as alterations in the expression of proteins related to cholesterol homeostasis. We also found that EB-3D and EB-3P lowered ChoKα protein levels. All these effects could be explained by the modulation of the AMP-activated protein kinase signalling pathway. We show that both inhibitors cause mitochondrial alteration and an endoplasmic reticulum stress response. EB-3D and EB-3P exert effects on ChoKα expression, AMPK activation, apoptosis, endoplasmic reticulum stress and lipid metabolism. Taken together, results show that EB-3D and EB-3P have potential anti-cancer activity through the deregulation of lipid metabolism.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Recent studies have shown the potential of magnetic hyperthermia in cancer treatments. However, the underlying mechanisms involved have not been yet fully described. In particular, the cell death ...related to magnetic hyperthermia observed in cultures incubated with low concentration of magnetic nanoparticles and under a low intensity alternating magnetic field, in which a macroscopic temperature rise is not observed, is still not understood. In the present study, we investigate the production of intracellular Reactive Oxygen Species (ROS) as a mechanism to induce cell death under these conditions. In this study, the production and influence of ROS on the viability of HepG2 human hepatoma cells (used as a model cell line) are analyzed under the application of variable magnetic fields using hyperthermia agents, such as biomimetic magnetic nanoparticles (BMNPs) mediated by magnetosome MamC protein from
Magnetococcus marinus
MC-1. The results show that intracellular ROS production increases up to ∼90% following upon the exposure of AMF to HepG2 cells containing BMNPs, which could determine the loss of cell viability (up to ∼40% reduction) without a significant rise in temperature. Such ROS production is linked to mitochondrial dysfunction caused by the application of AMF to cells containing BMNPs.
The production of Reactive Oxygen Species after exposure of HepG2 cells to alternating magnetic fields can explain the loss of cell viability in spite of a negligible increase in temperature.
Abstract
STUDY QUESTION
Which genes regulate receptivity in the epithelial and stromal cellular compartments of the human endometrium, and which molecules are interacting in the implantation process ...between the blastocyst and the endometrial cells?
SUMMARY ANSWER
A set of receptivity-specific genes in the endometrial epithelial and stromal cells was identified, and the role of galectins (LGALS1 and LGALS3), integrin β1 (ITGB1), basigin (BSG) and osteopontin (SPP1) in embryo–endometrium dialogue among many other protein–protein interactions were highlighted.
WHAT IS KNOWN ALREADY
The molecular dialogue taking place between the human embryo and the endometrium is poorly understood due to ethical and technical reasons, leaving human embryo implantation mostly uncharted.
STUDY DESIGN, SIZE, DURATION
Paired pre-receptive and receptive phase endometrial tissue samples from 16 healthy women were used for RNA sequencing. Trophectoderm RNA sequences were from blastocysts.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Cell-type-specific RNA-seq analysis of freshly isolated endometrial epithelial and stromal cells using fluorescence-activated cell sorting (FACS) from 16 paired pre-receptive and receptive tissue samples was performed. Endometrial transcriptome data were further combined in silico with trophectodermal gene expression data from 466 single cells originating from 17 blastocysts to characterize the first steps of embryo implantation. We constructed a protein–protein interaction network between endometrial epithelial and embryonal trophectodermal cells, and between endometrial stromal and trophectodermal cells, thereby focusing on the very first phases of embryo implantation, and highlighting the molecules likely to be involved in the embryo apposition, attachment and invasion.
MAIN RESULTS AND THE ROLE OF CHANCE
In total, 499 epithelial and 581 stromal genes were up-regulated in the receptive phase endometria when compared to pre-receptive samples. The constructed protein–protein interactions identified a complex network of 558 prioritized protein–protein interactions between trophectodermal, epithelial and stromal cells, which were grouped into clusters based on the function of the involved molecules. The role of galectins (LGALS1 and LGALS3), integrin β1 (ITGB1), basigin (BSG) and osteopontin (SPP1) in the embryo implantation process were highlighted.
LARGE SCALE DATA
RNA-seq data are available at www.ncbi.nlm.nih.gov/geo under accession number GSE97929.
LIMITATIONS, REASONS FOR CAUTION
Providing a static snap-shot of a dynamic process and the nature of prediction analysis is limited to the known interactions available in databases. Furthermore, the cell sorting technique used separated enriched epithelial cells and stromal cells but did not separate luminal from glandular epithelium. Also, the use of biopsies taken from non-pregnant women and using spare IVF embryos (due to ethical considerations) might miss some of the critical interactions characteristic of natural conception only.
WIDER IMPLICATIONS OF THE FINDINGS
The findings of our study provide new insights into the molecular embryo–endometrium interplay in the first steps of implantation process in humans. Knowledge about the endometrial cell-type-specific molecules that coordinate successful implantation is vital for understanding human reproduction and the underlying causes of implantation failure and infertility. Our study results provide a useful resource for future reproductive research, allowing the exploration of unknown mechanisms of implantation. We envision that those studies will help to improve the understanding of the complex embryo implantation process, and hopefully generate new prognostic and diagnostic biomarkers and therapeutic approaches to target both infertility and fertility, in the form of new contraceptives.
STUDY FUNDING/COMPETING INTEREST(S)
This research was funded by the Estonian Research Council (grant PRG1076); Horizon 2020 innovation grant (ERIN, grant no. EU952516); Enterprise Estonia (grant EU48695); the EU-FP7 Marie Curie Industry-Academia Partnerships and Pathways (IAPP, grant SARM, EU324509); Spanish Ministry of Economy, Industry and Competitiveness (MINECO) and European Regional Development Fund (FEDER) (grants RYC-2016-21199, ENDORE SAF2017-87526-R, and Endo-Map PID2021-127280OB-100); Programa Operativo FEDER Andalucía (B-CTS-500-UGR18; A-CTS-614-UGR20), Junta de Andalucía (PAIDI P20_00158); Margarita Salas program for the Requalification of the Spanish University system (UJAR01MS); the Knut and Alice Wallenberg Foundation (KAW 2015.0096); Swedish Research Council (2012-2844); and Sigrid Jusélius Foundation; Academy of Finland. A.S.-L. is funded by the Spanish Ministry of Science, Innovation and Universities (PRE2018-085440). K.G.-D. has received consulting fees and/or honoraria from RemovAid AS, Norway Bayer, MSD, Gedeon Richter, Mithra, Exeltis, MedinCell, Natural cycles, Exelgyn, Vifor, Organon, Campus Pharma and HRA-Pharma and NIH support to the institution; D.B. is an employee of IGENOMIX. The rest of the authors declare no conflict of interest.
En diversos estudios se ha mostrado la relación entre el nivel de forma física durante la infancia-adolescencia y el riesgo cardiovascular en la edad adulta. Dado que no se dispone de datos relativos ...al nivel de condición física de los adolescentes españoles, los objetivos de este estudio son:
a) determinar el nivel de condición física de los adolescentes españoles y establecer valores de referencia que puedan ser utilizados en el medio sanitario y educativo como indicadores de salud cardiovascular, y
b) conocer la proporción de adolescentes españoles que no alcanza valores de capacidad aeróbica indicativos de salud cardiovascular futura.
Se ha utilizado la batería EUROFIT modificada para evaluar la condición física de una muestra representativa de adolescentes españoles (n
=
2.859; 1.357 varones y 1.502 mujeres) procedente del estudio AVENA (Alimentación y Valoración del Estado Nutricional de los Adolescentes).
Se han obtenido los valores normativos de condición física de la población adolescente española. El rango del percentil 5 respecto a la capacidad aeróbica máxima (test de Course Navette) es de 2,0-3,3 y 1,4-1,9
paliers para varones y mujeres, respectivamente. Casi 1 de cada 5 adolescentes presenta riesgo cardiovascular futuro sobre la base de su capacidad aeróbica. Este subgrupo de adoles- centes mostró también una peor forma física que el resto de adolescentes en todas las pruebas físicas realizadas.
Los resultados obtenidos en el presente estudio permiten evaluar e interpretar correctamente el nivel de forma física de cualquier adolescente. Los resultados obtenidos indican la necesidad de mejorar el nivel de condición física de los adolescentes españoles.
Several studies have demonstrated that physical fitness in childhood and adolescence is related to cardiovascular risk in adulthood. Current data on the physical fitness of Spanish adolescents are not available. Therefore, the aims of this study were:
a) to assess the physical fitness of Spanish adolescents and establish reference values for use in health and educational settings as indicators of cardiovascular health, and
b) to determine the percentage of Spanish adolescents below the minimum level of aerobic fitness needed to guarantee future cardiovascular health.
The modified EUROFIT battery of tests was used to assess physical fitness in a representative sample of Spanish adolescents (n=2859; 1357 boys and 1502 girls) taking part in the AVENA (
Alimentación y Valoración del Estado Nutricional de los Adolescentes) study.
Standard parameters for the physical conditionof Spanish adolescents are reported in this study. The 5th percentile for maximum aerobic capacity (Course Navette test) ranged from 2.0-3.3 palier in boys and from 1.4-1.9 palier in girls. The findings indicate that, on the basis of aerobic fitness, approximately 20% of Spanish adolescents have an increased risk of future cardiovascular disease. This subgroup also performed poorly in all other tests of physical fitness used.
The results reported in this study enable the level of physical fitness in adolescents to be interpreted as an indicator of future cardiovascular health. They also indicate that the physical fitness of Spanish adolescents must be improved to help protect against cardiovascular disease in adulthood.
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