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

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 Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Lactologia Industrial; ArgentinaFil: Wolf, Irma Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico 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 Ingeniería Química; 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

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Variability of chemical composition and antioxidant activity of essential oils between Myrtus communis var. Leucocarpa DC and var. Melanocarpa DC

Essential oils (EOs) from several individuals of Myrtus communis L. (M. communis) growing in different habitats in Sardinia have been studied. The analyses were focused on four groups of samples, namely cultivated and wild M. communis var. melanocarpa DC, characterized by red/purple berries, and cultivated and wild M. communis var. leucocarpa DC, characterized by white berries. Qualitative and quantitative analyses demonstrated different EO fingerprints among the studied samples: cultivated and wild leucocarpa variety differs mainly from the melanocarpa variety by a high amount of myrtenyl acetate (>200 mg/mL and 0.4 mg/mL in leucocarpa and melanocarpa varieties respectively). Conversely, the wild group is characterized by a higher amount, compared with the cultivated species, of linalool (about 110 mg/mL and 20 mg/mL respectively), linalyl acetate (about 24 mg/mL and about 6 mg/mL respectively) whereas EOs of the cultivated plants were rich in pinocarveol-cis compared with wild plants (about 2 mg/mL and about 0.5 mg/mL respectively). Principal component analysis applied to the chromatographic data confirm a differentiation and classification of EOs from the four groups of M. communis plants. Finally, antioxidant activity of the studied EOs shows differences between the various categories of samples

A new approach to discriminate Rosmarinus officinalis L. plants with antioxidant activity, based on HPTLC fingerprint and targeted phenolic analysis combined with PCA

A new combined “untargeted” and “targeted” metabolomics approach to investigate Rosmarinus officinalis L. plants was proposed by using the merging of high-performance thin-layer chromatography (HPTLC) assay and liquid chromatography mass spectrometry (LC–MS) with principal component analysis (PCA). Rapid fingerprints of Rosmarinus extracts were obtained and compared by using HPTLC technique; targeted compounds were identified and quantified by LC–MS/MS (MRM), while the antioxidant capacity of extracts was evaluated by methods based on the ability to scavenge radicals such as ABTS+* and DPPH*, since the metabolite expression is generally correlated with the biological activity. By HPTLC fingerprints, a rapid visual comparison of secondary metabolites in Rosmarinus samples from different places of collection was obtained and PCA was performed providing an overview of the capacity of the HPTLC variables to discriminate samples. HPTLC fingerprint assisted by PCA results a reliable untargeted approach for the discrimination and classification of different samples on the basis of their geographical area, from west to east of Sardinia. The ‘targeted’ approach was performed by developing LC–MS/MS (MRM) method allowing the identification and quantification of 12 compounds belonging to the family of flavonoids and phenolic acids. The combination of quantitative data with PCA allowed to discriminate La Maddalena Rosmarinus sample from others, in a correlated way with antioxidant activities results, showing values significantly higher. A positive Pearson correlation between antioxidant activity measured with DPPH and ABTS and total phenolic contents, finally confirmed that antioxidant activity depends on phenolic amounts