Daniela Beghelli

Effect of supranutritional level of dietary α-tocopheryl acetate and selenium on rabbit semen

Abstract This research examined the effects of dietary α-tocopheryl acetate (50 or 200mg/kg diet) and selenium (Se, 0 or 0.5ppm) supplementation on motion characteristics, oxidative stability and fertilizing ability of rabbit spermatozoa, fresh and stored for 24h at 5°C. The higher amount of dietary α-tocopheryl acetate increased the level of Vitamin E in the fresh semen (1.75mmol/l versus 0.95mmol/l) and its oxidative stability (thiobarbituric acid reactive substances—TBARS 12.44nmol malondialdehyde/10 8 sperm versus 21.4nmol malondialdehyde/10 8 sperm). Dietary Se increased gluthatione peroxidase activity (GPx) in erythrocytes (285U/g Hb versus 207U/g Hb), seminal plasma (270U/l versus 190U/l) and spermatozoa (1338mU/10 9 sperm versus 1103mU/10 9 sperm), whereas it did not show any effect on α-tocopherol level and TBARS. No synergy between Vitamin E and Se was shown. Storage for 24h at 5°C increased the TBARS level in all the experimental groups. Neither live and acrosome reacted spermatozoa, nor kinetic parameters, nor fertility rate were modified by dietary supplementation.

CD4(-)CD8(-) T-cells in primary Sjögren's syndrome: association with the extent of glandular involvement.

OBJECTIVES Growing evidence suggests that IL-17-producing T cells, lacking both CD4 and CD8 molecules and defined as double negative (DN) cells, play a pivotal role in the pathogenesis of a number of systemic autoimmune disorders. We recently demonstrated that this T-cell subset is expanded in the peripheral blood (PB) of patients with primary Sjögrens syndrome (pSS), produces IL-17 and accumulates in minor salivary glands (MSGs). We aimed to investigate glandular and PB DN T cells in early pSS in order to verify a possible correlation with MSGs histological patterns and clinical parameters. METHODS Paired samples of PB mononuclear cells and MSGs from pSS patients were evaluated at the diagnosis by flow cytometry and immunofluorescence staining respectively. Histological analysis to identify histological scores, B/T cell segregation and the presence of germinal center (GC)-like structures was also performed. RESULTS In early stages of pSS, circulating DN T cells appear to be not yet expanded and inversely correlated with circulating CD4(+)Th17 cells. The number of infiltrating DN T cells were associated with extent of glandular involvement, presence of GC-like structures and dryness symptoms and were inversely correlated with circulating DN T cells. CONCLUSIONS Our findings suggest that DN T cells are actively involved in the pathogenic mechanisms leading to glandular dysfunction and damage in pSS and may play a role in ectopic lymphoneogenesis development occurring during the disease.

Quality of donkey meat and carcass characteristics

A study based on 15 entire donkey males was carried out to evaluate carcass quality and nutritional characteristics of meat obtained by these animals slaughtered at 15 months of age and a mean final body weight of 181kg. The meat had a low (2.02g/100g) fat content, an appreciable (22.8g/100g) protein content, and cholesterol content was 68.7mg/100g. Glycogen was also determined (0.45g/100g) within 12h of sampling. Potassium was the mineral with the highest content (343mg/100g), followed by phosphorus (212mg/100g), sodium (52mg/100g) and magnesium (24mg/100g). Donkey meat obtained from young animals can be considered a good alternative to other red meats and not only for the production salami, or other fermented meat products.

Donkey milk production: state of the art

Abstract Milk is one of the most common causes of food allergies among children under one year of age. No specific therapy exists for this allergy, and thus the only feasible response is to avoid assumption of milk and derived products. Studies conducted on the serum of children with hypersensitivity to milk have shown that caseins are the proteins with the greater allergenic potential. However, in some cases, children have also shown hypersensitivity to the β-lactoglobulines and to the α-lactal-bumins. When food intolerance is diagnosed in an infant, it is often necessary to impose a period of total parenteral feeding, followed by breast feeding, considered the most correct method of re-feeding. When human milk can not be given, alternative food sources must be sought. Clinical studies have demonstrated that donkey milk could substitute breast feeding in infants affected by severe Ig-E mediated milk allergies. In these subjects, donkey milk is not only useful, but also safer than other types of milk. In fact donkey milk composition in lipids (high levels of linoleic and linolenic acid) and proteins (low caseins content) is very close to human milk. Lysozyme content in donkey milk resulted to be very high (mean value 1.0 mg/ml) if compared to bovine (traces), caprine (traces) and human milk. The high lysozyme content of donkey milk may be responsible of the low bacterial count reported in literature and also makes this milk suitable to prevent intestine infections to infants. Among seroproteins, β-lactoglobulin and α-lactalbumin content in donkey milk was respectively 3.75 and 1.80 mg/ml and remained substancially the same during the different stages of lactation.

Physical and chemical characteristics of donkey meat from Martina Franca breed.

The rheological and chemical characteristics of meat obtained from 12 Martina Franca donkey males, slaughtered at 14months of age and a mean final body weight of 169kg were determined. Meat samples were taken four days post mortem from muscles Longissimus thoracis et lumborum and Biceps femoris, colorimetric parameters were measured to determine L(∗) (lightness), a(∗) (redness), b(∗) (yellowness) and chroma. The Longissimus was significantly lighter (P<0.05) compared to the Biceps femoris, with L(∗) indexes of 35.86 and 31.34, respectively. Fatty acid composition of the intramuscular fat showed a high content of polyunsaturated fatty acids (PUFAs) in both muscles, respectively 25.16g/100g total fatty acids in the Longissimus and 24.97g/100g total fatty acids in the Biceps femoris; oleic acid and palmitic acid were the two most abundant fatty acids in both muscles. The percentages of essential amino acids were higher in both muscles compared with the total amino acid content, respectively 52.88% in the Longissimus, and 51.26% in the Biceps femoris. The high level of unsaturation of the intramuscular fat resulted in a high ratio of unsaturated to saturated fat, and the total amount of essential amino acids, exceeding 50% of the total amino acids showed that donkey meat from a health point of view is a good alternative to traditional red meats.

Tenderness evaluation and mineral levels of llama (Lama glama) and alpaca (Lama pacos) meat

Tenderness and mineral levels were determined in the Longissimus thoracis taken from 20 llama and 30 alpaca males reared in Peru and slaughtered at 25 months of age. Mineral contents were determined using an inductively coupled plasma emission spectrometer. Tenderness evaluation was determined two and seven days post slaughter using a Warner-Bratzler shear force device. Potassium is the mineral with the highest content, with a significant difference (P<0.05) between the two species of camelids. The other mineral contents were, in decreasing order, phosphorus, sodium, magnesium and calcium, in addition to smaller percentages of zinc and iron. Shear force values determined seven days post slaughter were significantly (P<0.01) lower in both the species compared with the results obtained two days post slaughter.

Polar Constituents and Biological Activity of the Berry-Like Fruits from Hypericum androsaemum L.

Hypericum androsaemum, also known as Tutsan, is a small evergreen shrub common in the Mediterranean basin where it is traditionally used as diuretic and hepatoprotective herbal drug. This plant possesses the peculiarity to produce fleshy and berry-like fruits that ripen from red to shiny black. In the present work, the chemical constituents of methanolic extracts and infusions of red and black fruits were analyzed by HPLC, and correlated with their antioxidant properties which were evaluated by the DPPH, β-Carotene/linoleic acid, and hypochlorous acid tests. In addition, the red pigment of the fruit was isolated by column chromatography and structurally elucidated by NMR. Results showed that H. androsaemum fruits contain high amounts of shikimic and chlorogenic acids, while their color was given by a tetraoxygenated-type xanthone, reported for the first time in Hypericum species. The red berries infusion gave the highest content of total phenolic compounds, DPPH, and hypochlorous acid scavenging activity, and β-carotene bleaching. Cytotoxicity of the berries extracts on three human tumor cell lines (malignant melanoma, breast adenocarcinoma, and colon carcinoma) was evaluated by MTT assay, and relevant inhibition on colon carcinoma cells (IC50 value of 8.4 μg/mL) was found. Finally, the effects of red berries extract on the immune system were evaluated by peripheral blood mononuclear cell (PBMC) proliferation assay that revealed a strong stimulation on lymphocytes at low doses (0.4–6 μg/mL).

Hypoallergenic properties of donkey's milk: a preliminary study

Cows milk protein allergy (CMPA) is an abnormal immunological response to cow milk proteins, which results in IgE-mediated reactions. The therapeutic strategy to respond to CMPA envisages the total elimination of milk or the administration of cows milk substitutes. For this reason the use of milk from other mammalian species was tested. Among them, donkeys milk proved to be the best alternative in feeding infants affected by CMPA, since its chemical composition is comparable to human milk. In this work an in vitrostudy was performed in order to analyze the IgE reactivity to milk protein allergens from cow, donkey and goat. In particular, immunoblotting experiments using sera from milk-allergic and non-allergic adult volunteers were conducted with the aim of verifying the hypoallergenic property of donkeys milk. This study provided a preliminary evidence of the hypoallergenicity of donkeys milk when compared to bovine and goat milk. Considering the obtained results, it would be possible to develop a sensitive diagnostic method for CMPA detection, based on chromatographic and immunoblotting analysis.

IL-17-producing double-negative T cells are expanded in the peripheral blood, infiltrate the salivary gland and are partially resistant to corticosteroid therapy in patients with Sjögren’s syndrome

A small CD3+ T-cell population, that lacks both CD4 and CD8 molecules, defined as double negative (DN), is expanded in the peripheral blood of patients with systemic lupus erythematosus, produces IL-17 and accumulates in the kidney during lupus nephritis. Since IL-17 production is enhanced in salivary gland infiltrates of patients with primary Sjögrens syndrome (pSS), we aimed to investigate whether DN T cells may be involved in the pathogenesis of salivary gland damage. Fifteen patients with SS and 15 normal controls (NC) were enrolled. Peripheral blood mononuclear cells were stimulated with anti-CD3 antibody and cultured in presence or absence of dexamethasone (Dex). Phenotypic characterization was performed by flow cytometry in freshly isolated cells and after culture. Minor salivary glands (MSG) from pSS were processed for immunofluorescence staining. Total circulating DN T cells were increased in pSS compared to NC (4.7±0.4% vs 2.6±0.4%). NC and pSS freshly isolated DN T cells produce consistent amounts of IL-17 (67.7±5.6 in NC vs 69.2±3.3 in pSS). Notably, DN T cells were found in the pSS-MSG infiltrate. Dex was able to down-regulate IL-17 in vitro production in NC (29±2.6% vs 15.2±1.9% vs 13±1.6%) and pSS (49±4.8% vs 16±3.8% vs 10.2±0.8%) conventional Th17 cells and in DN T cells of NC (80±2.8% vs 3.8±2.1% vs 4.2±1.8%), but not of pSS (81±1.5% vs 85.4±0.8% vs 86.2±1.7%). DN T cells are expanded in pSS PB, produce IL-17 and infiltrate pSS MSG. In pSS, conventional Th17 cells are inhibited by Dex, but DN T cells appear to be resistant to this effect. Taken together, these data suggest a key role of this T-cell subset in the perpetuation of chronic sialoadenitis and eventually in pSS prognosis.

Somatic (CSS) and differential cell count (DCC) during a lactation period in ass’milk

Abstract Hypoallergenic properties of ass’s milk protein fractions have been recently confirmed, allowing ass’s milk to be considered as a valid substitute of the available hypoallergenic infant formulas. The objective of this study was to give a further contribution to the knowledge of ass’s milk safety and quality characteristics. A new procedure has been developed with a cytospin centrifuge in differential counts of milk somatic cells. Somatic cells count (SCC), differential somatic cells count (DCC) and cultural examinations have been carried out in 62 milk samples collected from 11 asses at three different stages of lactation. Four major cells populations had been identified in ass’s milk too: lymphocytes (Ly), monocytes/macrophages (MA), polymorphonuclear neutrophils (PMNL), and epithelial cells (CE). The patterns of these cells have been discussed in comparison with cells found in dairy cows and ewes milk. In conclusion, a reproducible standard procedure has been developed to determine cell count of ass’s milk.