In vitro and in vivo studies of Cucurbita pepo L. flowers: chemical profile and bioactivity.

Edible flowers consumption has increased in recent years due to their rich content of healthy phytochemicals. The aim of this study was to analyse the chemical profile of Cucurbita pepo L. flowers, and to explore their antioxidant and hypoglycaemic properties. Moreover, in order to assess in vivo effects, biochemical analysis, Reactive Oxygen Metabolites (d-ROMs) and Biological Antioxidant Potential (BAP) tests were performed on mice serum. High Performance Liquid Chromatography-Diode Array Detection (HPLC-DAD) analyses revealed the presence of (+)-catechin, (-)-epicatechin, rutin, and syringic acid as main constituents. 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and Ferric Reducing Antioxidant Power (FRAP) tests showed interesting results. The extract exhibited the strongest inhibitory effect on α-glucosidase (IC50 of 144.77 µg/mL). In vivo results confirmed the hypoglycaemic effects, also affecting lipid metabolism but did not revealed benefits on ROS production. These results may add some information supporting the use of C. pepo flowers as functional foods and/or nutraceuticals.

Influence of dietary hydrogenated palm oil supplementation on serum biochemistry and progesterone levels in dairy goats.

The aim of this research was to investigate the influence of hydrogenated palm oil (HPO) added to a dairy goat diet on serum biochemistry and progesterone levels. Thirty pregnant Cilentana dairy goats were equally divided into 2 groups (control [CTR] and HPO groups). After kidding, concentrated feed for both groups was gradually increased up to 400 g/(animal·d), and the HPO group received 50 g/(animal·d) of HPO. Supplementation with HPO significantly increased cholesterol levels (mg/dL, 63.80 vs. 54.68 at 30 d, P ≤ 0.05; 78.20 vs. 58.00 at 60 d, P ≤ 0.05; 83.80 vs. 57.83 at 120 d, P ≤ 0.01) compared with the CTR group although no significant differences were detected for liver and kidney function indicators. Moreover, other biochemical parameters were not affected by HPO supplementation thus suggesting no change occurred in lipid and protein metabolism. Furthermore, a significant correlation was found between progesterone levels and serum cholesterol (r = 0.65, P ≤ 0.01) although these were not significantly higher in HPO supplemented goats. The dose and time of HPO supplementation appears critical as regards assessing the limits between the risks and benefits of HPO supplementation in dairy goats. At the tested dose, HPO was well tolerated by the animals and may represent a useful tool to increase energy availability during highly demanding periods.

Key Role of Cytochrome C for Apoptosis Detection Using Raman Microimaging in an Animal Model of Brain Ischemia with Insulin Treatment.

Brain ischemia represents a leading cause of death and disability in industrialized countries. To date, therapeutic intervention is largely unsatisfactory and novel strategies are required for getting better protection of neurons injured by cerebral blood flow restriction. Recent evidence suggests that brain insulin leads to protection of neuronal population undergoing apoptotic cell death via modulation of oxidative stress and mitochondrial cytochrome c (CytC), an effect to be better clarified. In this work, we investigate on the effect of insulin given intracerebroventricular (ICV) before inducing a transient global ischemia by bilateral occlusion of the common carotid arteries (BCCO) in Mongolian gerbils (MG). The transient (3 min) global ischemia in MG is observed to produce neurodegenerative effect mainly into CA3 hippocampal region, 72 h after cerebral blood restriction. Intracerebroventricular microinfusion of insulin significantly prevents the apoptosis of CA3 hippocampal neurons. Histological observation, after hematoxylin and eosin staining, puts in evidence the neuroprotective role of insulin, but Raman microimaging provides a clearer insight in the CytC mechanism underlying the apoptotic process. Above all, CytC has been revealed to be an outstanding, innate Raman marker for monitoring the cells status, thanks to its resonant scattering at 530 nm of incident wavelength and to its crucial role in the early stages of cells apoptosis. These data support the hypothesis of an insulin-dependent neuroprotection and antiapoptotic mechanism occurring in the brain of MG undergoing transient brain ischemia. The observed effects occurred without any peripheral change on serum glucose levels, suggesting an alternative mechanism of insulin-induced neuroprotection.

Detection of buffalo milk adulteration with cow milk by capillary electrophoresis analysis.

The addition of cow milk during the production of buffalo mozzarella is a common fraud in dairy industries because of the lower price and greater availability of cow milk throughout the year. The aim of this study was to develop a new, rapid, and robust capillary electrophoresis method for detecting and quantifying cow milk in buffalo milk by exploiting cow α-lactalbumin as a marker of adulteration. In particular, a linear calibration curve was generated, using a training set of calibrators consisting of 7 series of 17 buffalo/bovine whey mixtures, obtained after casein precipitation, with increasing percentages of cow whey. The capillary electrophoresis method showed high linearity (R2 = 0.968), repeatability [relative standard deviation (RSD) = 2.11, 3.02, 4.38, and 1.18%, respectively for 5, 10, 20, and 50% of buffalo/bovine whey mixtures], and intermediate precision (RSD = 2.18, 2.49, 5.09, and 3.19%, respectively, for 5, 10, 20, and 50% buffalo/bovine whey mixtures). Moreover, the minimum amount of detectable fraudulent cow milk was 1%, and the limit of quantification was 3.1%.

Gemcitabine-loaded liposomes: rationale, potentialities and future perspectives.

This review describes the strategies used in recent years to improve the biopharmaceutical properties of gemcitabine, a nucleoside analog deoxycytidine antimetabolite characterized by activity against many kinds of tumors, by means of liposomal devices. The main limitation of using this active compound is the rapid inactivation of deoxycytidine deaminase following administration in vivo. Consequently, different strategies based on its encapsulation/complexation in innovative vesicular colloidal carriers have been investigated, with interesting results in terms of increased pharmacological activity, plasma half-life, and tumor localization, in addition to decreased side effects. This review focuses on the specific approaches used, based on the encapsulation of gemcitabine in liposomes, with particular attention to the results obtained during the last 5 years. These approaches represent a valid starting point in the attempt to obtain a novel, commercializable drug formulation as already achieved for liposomal doxorubicin (Doxil(®), Caelyx(®)).

Improved in vitro and in vivo collagen biosynthesis by asiaticoside-loaded ultradeformable vesicles.

The potentiality of ultradeformable vesicles as a possible topical delivery system for asiaticoside, a natural compound obtained from Centella asiatica was evaluated, because this compound exhibits collagen biosynthesis promoting activity. Ultradeformable vesicles were prepared by the extrusion technique; these vesicles were composed of Phospholipon 100 and different molar fractions of sodium cholate as the edge activator. The physicochemical properties of the ultradeformable vesicles were investigated through differential scanning calorimetry and light scattering techniques. The potential cyctotoxicity and biological activity of asiaticoside-loaded ultradeformable vesicles were evaluated on primary human dermal fibroblast cells by determining the extracellular lactic dehydrogenase activity, the cellular viability and the biosynthetic production of collagen. In vitro permeation experiments through human stratum corneum and epidermis membranes were also carried out. Ultradeformable vesicles having sodium cholate molar fraction of 0.2 proved to be the most suitable topical carriers for asiaticoside. A sodium cholate content of >0.2 was observed to be cytotoxic probably due to its co-existence with other lipid aggregates, an example being mixed micelles. Asiaticoside-loaded ultradeformable vesicles with a sodium cholate molar fraction of 0.2 elicited the greatest degree of collagen biosynthesis in human fibroblasts. Ultradeformable vesicles provided the greatest in vitro skin permeation of asiaticoside showing a 10-fold increase with respect to the free drug solution and favoured an increase in in vivo collagen biosynthesis. Ultradeformable vesicles are therefore suitable carriers for the pharmaceutical and cosmetic application of the natural agent asiaticoside.