27 research outputs found
Le Cellule endoteliali del cordone ombelicale hanno un destino cellulare che dipende dal sesso del feto
Aims: The endothelial cells (EC) play a crucial role in the regulation of a broad range of many physiological and pathological processes. The aims of this work are to evaluate possible gender differences in EC obtained by male and female newborn umbilical cord in cell fate and in the ability of cell survival in starvation an inducer of autophagy.
Methods: HUVECs (human umbilical vein endothelial cells) were isolated as previously described by Cheung L et al. and umbilical cord were provided by the department of Pharmacology, Gynecology, and Obstetrics by University of Sassari. The cell proliferation was evaluated with MTS assay kit and dimension and cell shape were evaluated with Labworks sofware. Moreover we determinate H2O2 LDH and PAI-1 concentration with commercial colorimetric assay kit respectively in cell supernatants and, eNOS, Beclin1, LC3, AKT, mTOR expression by western blotting on cell lysates according to the instructions provided by Cell Signalling Tecnology assay kit and nitrite quantification by Griess reaction on culture supernatants. The experiments were performed with the 5 passage.
Results: Basically, no significant differences are observed in cell shape and dimension, LDH, PAI-1 and nitrite and nitrate production levels and espression of eNOS. But basal male cells have a higher expression of Beclin1, an autophagic marker, while the expression of AKT and its activity as mTOR expression are significantly higher in female than in male cells. After 36 hours of starvation, as expected both cell types are more autophagic, but male cells shows significantly higher expression of ratio LC3II/LC3I, an autophagic activity indicator. Finally, both cell types have lower AKT and mTOR espression and both cells show no AKT activity.
Conclusion: When intracellular nutrients are limited as during starvation, eukaryotic cells can trigger a pathway of survival called autophagy. Here, it is evident that male and female endothelial cells have different responses
Statistical Analysis Applied to the Production of Mirto Liqueur
Preparation of myrtle liqueur through ethanol-based extraction is a widely employed methodology. Nevertheless, optimization of existing processes is possible, especially through a modern statistical multivariate approach. In this context, a Design of Experiments (DoE) approach was used to quantitatively assess for the first time the effect of the time, ethanol concentration, temperature, and the ratio between the Myrtus communis berries' weight and the extractant volume (v/w) on the amounts of anthocyanins, volatile compounds and dry residues in the liqueur. The kinetic profile relative to the volatile fraction variation during the process was described by gas chromatography (GC), while spectrophotometric analysis allowed quantification of the total anthocyanins and total polyphenols. Multiple response analysis showed that the maximum efficiencies in terms of the considered parameters (desirability function) were reached by setting the temperature to 25 degrees C and the ethanol percentage to 96% after 20 days of processing. Some hints as to the chemical instability and not negligible sensitivity of anthocyanins in relation to the experimental conditions for longer extraction times were also observed. The statistical model represents a novel tool for industrial production of myrtle liqueur
Profiling of the Bioactive Compounds in Flowers, Leaves and Roots of Vinca sardoa
Vinca sardoa (Stern) Pignatti (Apocynaceae) is largely distributed in Sardinia where it is considered a typical endemism. Since the plant is used in traditional folk medicine, the aim of the present work was to identify and quantify the polyphenolic metabolites, due to the well-known importance of polyphenolic compounds, as well as to evaluate antioxidant activity in different parts of the plant. The compounds were identified and determined in the methanol extracts of leaves, flowers, and roots by developing different LC-MS/MS methods. The obtained data show that leaves possess the highest amount of polyphenols, in particular quinic acid (3401 mg/100 g), chlorogenic acid (1082 mg/100 g), caffeoylquinic acid isomer 1 (190 mg/100 g), and robinin (633 mg/100 g). Likewise, antioxidant tests showed that leaves possess the main radical scavenging activities in both ABTS (49.19 ± 3.41 μg/mL, 30.88 ± 3.04 μg/mL at time zero and after 50 min, respectively) and DPPH assays (223.97 ± 30.81 μg/mL, 109.52 ± 12.89 μg/mL at time zero and after 30 min, respectively). Taking into account that leaves differed most from flowers and roots in the content of caffeoylquinic acid and chlorogenic acid, of which antioxidant properties are widely recognized, it is reasonable to assume that these two compounds are involved in the differences described. The relationship between the high polyphenolic content and interesting antioxidant activities, justifies its use in ethnobotany and may be suggest a use of this specie, after removal of the alkaloid fraction, in the pharmaceutical, phytotherapy, and cosmetic industries
Incorporation of probiotic bacteria (Lactobacillus acidophilus and Bifidobacterium ssp.) in Argentinean ovine cheese
The market of cheeses from ewe’s milk has been growing steadily in Argentina. The nutritional benefits of these products can be enhanced by adding probiotic cultures. In the present study, the survival of a mix of probiotic microorganisms (Lactobacillus acidophilus La-5 and Bifidobacterium animalis ssp. lactis Bb12) in a semi-hard ovine cheese, manufactured with a technology previously developed in our institute was evaluated. Besides, the effect of its incorporation on the chemical composition and ripening parameters of cheeses, including the fatty acid composition and CLA production, was investigated. Experimental cheeses made with Bb12 and La-5 retained counts of the probiotic strains at the required therapeutic level (107 CFU g−1 ). No significant differences were detected between experimental and control cheeses with respect to the different parameters evaluated: gross composition, nitrogen fractions, lipolysis, fatty acids profiles of cheese fat including CLA, and volatile fraction. Results demonstrated that the ovine cheese matrix seems to be suitable for delivery of probiotic microorganisms.Fil: Perotti, Maria Cristina. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Santa Fe. Instituto de Lactologia Industrial; ArgentinaFil: Wolf, Irma Veronica. Consejo Nacional de Investigaciones CientÃficas y Técnicas. Centro CientÃfico Tecnológico Santa Fe. Instituto de Lactologia Industrial; ArgentinaFil: Addis, Margherita. AGRIS Sardegna. Dipartamento per la ricerca nelle produzioni animali (DIRPA), Sardegna; ItaliaFil: Comunian, Roberta. AGRIS Sardegna. Dipartamento per la ricerca nelle produzioni animali (DIRPA), Sardegna; ItaliaFil: Paba, Antonio. AGRIS Sardegna. Dipartamento per la ricerca nelle produzioni animali (DIRPA), Sardegna; ItaliaFil: Meinardi, Carlos Alberto. Universidad Nacional del Litoral. Facultad de IngenierÃa QuÃmica; Argentin
Human umbilical endothelial cells (HUVECs) and sex differences
HUVECs are worldwide used to study the endothelial physiology and pathology that might be involved in sex and gender differences detected at the cardiovascular level. The present work characterised the phenotype of HUVECs in terms of morphology, proliferative and migratory capacity and in the gene expression of oestrogen and androgen receptors and nitric oxide synthase 3 (NOS3) to evaluated if they are sexually dimorphic. Moreover, autophagic process was analysed in male and female HUVECs (MHUVECs and FHUVECs), as autophagy is influenced by sex. Umbilical cords were obtained from healthy, normal weight, male and female neonates born to healthy non-obese and non-smoking women. HUVECs morphology was analysed by electron microscopy, and their function was investigated by proliferation, viability, wound healing and chemotaxis assays. Real-time PCR was used to evaluate gene expression for oestrogen and androgen receptors and for NOS3, while the expression of the primary molecules involved in autophagic process [(Akt, the mammalian target of rapamycin (mTOR), beclin-1 and microtubule-associated protein 1 light chain 3 (LC3)] and NOS3 were analysed by western blotting. FHUVECs showed significantly higher proliferation and migration rate, and NOS3 mRNA and protein expression than MHUVECs. Conversely, beclin-1 and the LC3-II/LC3-I ratio were higher in MHUVECs than in FHUVECs, indicating a higher autophagy in male cells as also indicated by ultrastructural analysis showing a buildup of autophagic vacuoles at different stages in MHUVECs. The expression of oestrogen and androgen receptor genes, the protein expression of Akt, mTOR, and cellular size and shape were not influenced by sex. Male and female neonates did not differ in body weight, but the weight of male babies was positively associated with the weight of the mother, suggesting that the weight of the mother may exert a different influence on male and female babies. Our findings indicate that sex differences exist from prenatal life and are parameter- specific, suggesting that a better quality of the research on the endothelium in vitro can be obtained by analyzing HUVECs of both sexes as well as its translational value. Moreover, the sex differences observed in HUVECs could help the diseases of adulthood because endothelial dysfunction has a key role in cardiovascular diseases, diabetes mellitus, neurodegeneration and immune diseases
Human umbilical endothelial cells (HUVECs) have a sex: characterisation of the phenotype of male and female cells
Background: Human umbilical endothelial cells (HUVECs) are widely used to study the endothelial physiology and pathology that might be involved in sex and gender differences detected at the cardiovascular level. This study evaluated whether HUVECs are sexually dimorphic in their morphological, proliferative and migratory properties and in the gene and protein expression of oestrogen and androgen receptors and nitric oxide synthase 3 (NOS3). Moreover, because autophagy is influenced by sex, its degree was analysed in male and female HUVECs (MHUVECs
and FHUVECs).
Methods: Umbilical cords from healthy, normal weight male and female neonates born to healthy non-obese and non-smoking women were studied. HUVEC morphology was analysed by electron microscopy, and their function
was investigated by proliferation, viability, wound healing and chemotaxis assays. Gene and protein expression for
oestrogen and androgen receptors and for NOS3 were evaluated by real-time PCR and Western blotting, respectively, and the expression of the primary molecules involved in autophagy regulation [protein kinase B (Akt), mammalian target of rapamycin (mTOR), beclin-1 and microtubule-associated protein 1 light chain 3 (LC3)] were detected by Western blotting.
Results: Cell proliferation, migration NOS3 mRNA and protein expression were significantly higher in FHUVECs
than in MHUVECs. Conversely, beclin-1 and the LC3-II/LC3-I ratio were higher in MHUVECs than in FHUVECs, indicating
that male cells are more autophagic than female cells. The expression of oestrogen and androgen receptor genes and
proteins, the protein expression of Akt and mTOR and cellular size and shape were not influenced by sex. Body weights of male and female neonates were not significantly different, but the weight of male babies positively correlated with the weight of the mother, suggesting that the mother’s weight may exert a different influence on male and female babies.
Conclusions: The results indicate that sex differences exist in prenatal life and are parameter-specific, suggesting that HUVECs of both sexes should be used as an in vitro model to increase the quality and the translational value of research. The sex differences observed in HUVECs could be relevant in explaining the diseases of adulthood because endothelial dysfunction has a crucial role in the pathogenesis of cardiovascular diseases, diabetes mellitus, neurodegeneration and
immune disease.</br
Identification of cork characters for phenotypic selection
Global change effects can determine major changes in species distribution and productivity. In the Mediterranean region of the severity of hot and dry periods is growing and an increased frequency of extreme events and a major vulnerability of natural ecosystems is evident. Cork oak (Quercus suber L.) is widely spread in the West Mediterranean region and its economic and social is important
Statistical Analysis Applied to the Production of Mirto Liqueur
Preparation of myrtle liqueur through ethanol-based extraction is a widely employed methodology. Nevertheless, optimization of existing processes is possible, especially through a modern statistical multivariate approach. In this context, a Design of Experiments (DoE) approach was used to quantitatively assess for the first time the effect of the time, ethanol concentration, temperature, and the ratio between the Myrtus communis berries’ weight and the extractant volume (v/w) on the amounts of anthocyanins, volatile compounds and dry residues in the liqueur. The kinetic profile relative to the volatile fraction variation during the process was described by gas chromatography (GC), while spectrophotometric analysis allowed quantification of the total anthocyanins and total polyphenols. Multiple response analysis showed that the maximum efficiencies in terms of the considered parameters (desirability function) were reached by setting the temperature to 25 °C and the ethanol percentage to 96% after 20 days of processing. Some hints as to the chemical instability and not negligible sensitivity of anthocyanins in relation to the experimental conditions for longer extraction times were also observed. The statistical model represents a novel tool for industrial production of myrtle liqueur
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