The Aggregated and Micellar Forms of ß-Casein Purified from Donkey and Bovine Milk Present Potential as Carriers for Bioactive Nutritional Compounds.

In recent years, there has been a growing interest in the pure casein fraction of milk protein, particularly ß-casein due to its physicochemical properties as well as its bio- and techno-functional properties. The utilization of self-assembled ß-caseins from bovine origin as nanocarriers for the delivery of nutraceutical compounds or drugs has increased dramatically. Concerning ß-caseins from other milk sources, the use of hypoallergenic donkey ß-caseins as a potential delivery vehicle for nutraceutical hydrophobic compounds is beginning to generate interest. The present review deals with casein micelles models, bovine and donkey ß-casein molecular structures, as well as their physical-chemical properties that account for their exploitation in nutraceutics and pharmaceutics. This review work suggests the possibility of developing delivery systems for hydrophobic bioactive compounds using ß-casein purified from hypoallergenic donkey milk, highlighting the potential of this protein as an innovative and promising vehicle for enhancing the enrichment and bioavailability of various bioactive substances in food products.

Salt content of prepacked cereal-based products and their potential contribution to salt intake of the Italian adult population: Results from a simulation study.

BACKGROUND AND AIMS: High sodium intake is one of the main risk factors for noncommunicable diseases, and its consumption should be reduced. This study aimed to simulate changes in the daily salt intake of the Italian adult population based on consumption scenarios of prepacked cereal-based foods sold in Italy. METHODS AND RESULTS: Information on food packages was retrieved from 2893 cereal-based products. Potential changes in salt intake were simulated based on food consumption scenarios that consider the daily consumption of cereal-based products suggested in the Italian Dietary Guidelines and their current daily consumption by Italian adults. The highest salt content was retrieved in bread (median, 25th-75th percentile: 1.3, 1.1-1.4 g/100 g) and bread substitutes (1.8, 1.0-2.2 g/100 g). If the suggested daily amounts were consumed, bread would contribute to 44% of the 5 g salt/day target, whereas bread substitutes, breakfast cereals, biscuits and sweet snacks would marginally contribute (1-2%). Compared to bread with median salt content, a -44% and +10% salt intake would be observed if products within the first and the last quartile of salt content were chosen, respectively. However, considering the actual intake of Italian consumers, bread would cover 25% and bread substitutes 7% of the daily salt target. CONCLUSION: Food labels have a pivotal role and efforts are required to encourage consumers to use them to make healthy choices. Moreover, these results may contribute to setting sodium benchmarks in cereal-based products and encourage the food industry to reduce the salt content in the products.

Effects of different steam injection conditions on cappuccino's nutritional profile.

The quality parameters of cappuccinos prepared with pasteurized milk or ultra-high-temperature milk steam-injected at different temperatures by a professional coffee machine have been assessed. In particular, the protein profile, the content of vitamins and lactose, the lipid peroxidation process, and the involvement of milk proteins in the foam formation were evaluated. The nutritional quality of milk seems not affected by the steam injection treatment carried out at a temperature of 60-65 °C, but at higher temperatures a decrement of lactoperoxidase, vitamin B6 and folic acid was observed. The milk used in cappuccino preparation is very important: pasteurized milk can form a more consistent and lasting foam with respect to ultra-high-temperature milk because of the presence of ß-lactoglobulin and lactoferrin, both playing an important role in the foam formation and stability. This work would provide additional information to the coffee industry for the preparation of high nutritional and organoleptic quality cappuccinos.

Nutritional Properties of Rice Varieties Commonly Consumed in Italy and Applicability in Gluten Free Diet.

Gluten-free diets are often characterized by an inadequate intake of nutrients and are generally monotonous for the limited number of products celiac patients can use. As rice is the most used cereal by celiac consumers, studying rice varieties nutritional characteristics is of interest to manage diet quality and variety. Proteins, total carbohydrates and amylose content of six rice varieties (Ribe, Vialone Nano, Carnaroli, Arborio, Basmati, and Fragrance) were analyzed. Analyses were performed in raw products and after boiling, stewing, and microwaving. A decrease of proteins and total carbohydrates amount was observed in cooked rice. The same was reported for amylose content with boiling showing the highest loss (average retained amylose 53%). Considering amylose percentage with respect to total carbohydrates, each variety showed either an increase or a decrease depending on cooking method. The highest values were obtained with stewing above all for Basmati rice and Arborio rice. However, exceptions can be underlined as Carnaroli rice, showing the highest percentage when boiled. In this context, nutritional characteristics of cooked rice varieties appear to be of great importance to increase specific nutritional knowledge to better manage gluten-free diets.

Vitamins in Human and Donkey Milk: Functional and Nutritional Role.

BACKGROUND: Whole milk is a good source of all the nutrients, and it also contains a sufficient number of vitamins to permit regular the growth of the neonate. Dairy cow milk can create allergy in infants less than 12 months old because of the high caseins and ß-lactoglobulin content. In these circumstances, donkey milk can represent a good replacement for dairy cows' milk in children affected by Cow Milk Protein Allergy (CMPA) because of its close chemical composition with human milk, mainly due to its low protein and low mineral content. Milk vitamin content is highly variable among mammalian species and it is strictly correlated with the vitamin status and the diet administered to the mother. Fat-soluble vitamins content in donkey milk is, on average, lower compared to ruminants' milk, while vitamin C content determined in donkey milk is higher compared to dairy cows' milk, showing a great similarity with human milk. In donkey milk, the content of vitamins of the B-complex such as thiamine, riboflavin, niacin, pyridoxine, and folic acid is higher compared to human milk. The use of donkey milk as a new functional food must be further evaluated in interdisciplinary clinical trials in which pediatricians, dietitians, and food scientists must be involved to deepen the knowledge about the positive health impact of donkey milk in different sensitive people, especially children and the elderly.

Faecal proteome in clinically healthy dogs and cats: Findings in pooled faeces from 10 cats and 10 dogs.

BACKGROUND: In the scientific literature, there are only a few manuscripts available on small animal faecal proteomics. METHODS: In the present pilot study, this evaluation was performed using pooled faecal samples from 10 clinically healthy dogs and, for the first time, in 10 clinically healthy cats by mean of two-dimensional electrophoresis followed by liquid chromatography-tandem mass spectrometry. RESULTS: Our results showed the presence of nine (albumin, alkaline phosphatase, chymotrypsin-C-like, cytosol aminopeptidase, elastase-3B/proteinase E, immunoglobulins and nuclear pore membrane glycoprotein 210) and 14 (albumin, caspase recruitment domain-containing protein, chymotrypsin-like, deleted in malignant brain tumours 1 protein-like, hypothetical protein LOC107375, immunoglobulin, kallikrein-1, superoxide dismutase, transthyretin precursor, interstitial collagenase-like) different proteins in canine and feline faeces, respectively. CONCLUSION: These preliminary findings document the presence of a range of proteins in the faeces of apparently healthy dogs and cats and may serve as a basis for larger, prospective studies to establish reference proteomic data against which diseased populations can be compared.

Rheological Properties and Growth Factors Content of Platelet-Rich Plasma: Relevance in Veterinary Biomedical Treatments.

Platelet-rich plasma (PRP) is a nontransfusional hemocomponent, considered as a powerful concentrate of growth factors (GFs) therapeutically used to stimulate tissue regeneration. The use of autologous PRP, as the patient's own biological material, for therapeutic purposes represents a safe and effective alternative to conventional treatments in both human and veterinary medicine. The aim of this study was the characterization of canine PRP from rheological and biological points of view. Thus, a characterization of the viscoelastic properties of the PRP systems was performed in order to clarify the influence of different calcium concentrations, in the presence of autologous thrombin-rich solution, on the PRP gels' mechanical properties, from which the applicability of these systems in biomedical treatments is strongly dependent. Then, an evaluation of the content of GFs in PRP, activated or not with thrombin, and stored at different temperatures (37 °C and -20 °C) was performed over time, outlining, for the first time, the importance of the effect of physiological temperature (37 °C) on the production of GFs. A clinical case study conducted in a dog with a complete rupture of the common calcaneal tendon (Achilles tendon) confirmed the relevance of this hemocomponent in the daily veterinary clinical activity and the potential translational value for human health.

Comparison of Carcass and Meat Quality Obtained from Mule and Donkey.

The aim of this study was to compare the carcass characteristics and the chemical and physical parameters of the meat produced by 10 male crossbred donkeys and 10 male mules slaughtered at 16 ± 1 years of age. The carcass weight and dressing percentage were significantly (p < 0.05) higher in mules. Samples of the muscle Longissimus thoracis (LT) were analyzed. Donkey meat showed a higher fat content, while the glycogen content was higher in the mule LT. The total collagen content was higher in the mule LT; in this muscle, the shear force values were higher compared to the donkey LT. The lightness parameter (L*) was lower in the mule LT, while the redness index (a*) was higher in the mule LT; this muscle showed a higher content of iron, while zinc was higher in the donkey muscle LT. The donkey LT muscle showed a higher content of essential amino acids (52.2%) compared to the mule LT (50.1%). The results obtained demonstrated that the chemical and physical traits of mule and donkey meat were similar to those of other kinds of equid meat.

Proteomic characterization of kefir milk by two-dimensional electrophoresis followed by mass spectrometry.

Kefir is a type of fermented milk obtained thanks to the introduction of "kefir grains" in mammalian milk. Kefir grains consist of lactic and acetic acid bacteria and yeasts in alternative proportions that are held together by a matrix of complex sugars known as "kefiran." Thanks to the fermentative process, the kefir milk is rich in nutraceutical substances such as amino acids, vitamins, and mineral salts. The most valuable compounds of kefir fermentation are mainly lactic acid, exopolysaccharides, and bioactive peptides, the resulting products of proteolytic release from milk proteins (caseins and whey proteins). Among the nutraceutical properties of kefir are antimicrobial and antitumor activity, immunomodulating effect, and cholesterol-lowering effect. Therefore, in light of these intriguing properties of kefir milk, in this work, a proteomic analysis, by two-dimensional electrophoresis followed by mass spectrometry, has been performed. As a result, milk-derived polypeptides were identified in commercial kefir milk from organic farming. In particular, polypeptides deriving from κ-, αs1 -, and αs2 -caseins that may have potentially beneficial effects on human health have been detected.

Dietary Intake of Vitamin D from Dairy Products Reduces the Risk of Osteoporosis.

BACKGROUND: Vitamin D and calcium are important dietary compounds that affect bone mass, even if other minerals (potassium, zinc, etc.) and vitamins (A, C and K) are also involved. Vitamin D and certain minerals, in fact, play an important role in calcium homeostasis and calcium absorption. Hip fracture incidence is higher in Europe and the United States, where calcium is frequently included in the human diet; while the occurrence of these fractures is lower in developing countries, where diets are often poor in calcium. This condition is named the "calcium paradox", and may be partially explained by phosphate toxicity, which can negatively affect mineral metabolism. It is important to maintain correct dietary calcium-phosphate balance in order to have a healthy life, reducing the risk of osteoporotic fractures in older people. Vitamin D can also act as a hormone; vitamin D2 (ergocalciferol) is derived from the UV-B radiation of ergosterol, the natural vitamin D precursor detected in plants, fungi, and invertebrates. Vitamin D3 (cholecalciferol) is synthesized by sunlight exposure from 7-dehydrocholesterol, a precursor of cholesterol that can also act as provitamin D3. Dietary intake of vitamin D3 is essential when the skin is exposed for short periods to ultraviolet B light (UV-B), a category of invisible light rays such as UV-A and UV-C. This can be considered the usual situation in northern latitudes during the winter season, or the typical lifestyle for older people and/or for people with very white delicate skin. The actual recommended daily intake of dietary vitamin D is strictly correlated with age, ranging from 5 µg for infants, children, teenagers, and adults-including pregnant and lactating women-to 15 µg for people over 65 years.

Possible Role of Vitamin D in Celiac Disease Onset.

Beside skeletal system maintenance and protection, possible extra-calcium roles of vitamin D have been recently described. In particular, studies have investigated possible roles of vitamin D as a key modulator of inflammation and immune mechanisms and of the intestinal mucosa barrier. In this regard, vitamin D has been considered as a factor that affects different conditions such as immune-mediated diseases. The new emerging role of vitamin D and its involvement in immune modulation has led it to be considered as a possible key factor involved in celiac disease (CD) onset. CD is a chronic immune-mediated enteropathy of the small intestine that is triggered by dietary gluten protein exposure in individuals who are genetically predisposed. However, along with gluten, other environmental factors are also involved in CD onset. The renewed interest in a molecule that offers great possibilities for new roles has led to an increase in studies, although there remains a lack of studies aimed at contextualizing the role of vitamin D on CD. This review aims to define the possible role of vitamin D in CD onset as it is presently understood, taking into account potential links among vitamin D, the immune system and CD.

Microbiological Assessment of the Quality of Some Commercial Products Marketed as Lactobacillus crispatus-Containing Probiotic Dietary Supplements.

In the last decade, many authors have reported low viability for probiotic products. Investigators commonly find they are not meeting claimed active counts and/or incorrect species and/or strains have been identified. We have therefore decided to verify viability, the real dose and species correspondence in nine probiotic products (seven nutritional supplements and two medical devices) collected from the Italian and French markets claiming to contain at least one strain of L. crispatus among the different species/strain included in the formulation. In fact, the medical relevance of L. crispatus strains has recently grown., as evaluating the possible dominance clusters typical of the vaginal microbiota, the Community State Type I, the one dominated by L. crispatus, appears to be "protective" in terms of infections, fertility and gestational duration of pregnancy. The results obtained demonstrate the generally poor quality of probiotics. Out of nine products, only two definitely contained viable Lactobacilluscrispatus cells with a daily dose of at least 1 × 109 CFU/g and with an acceptable correspondence with what is declared on the label. Among these two, only one was found to be formulated with a strain (M247) that has been scientifically documented.

A comparison among ß-caseins purified from milk of different species: Self-assembling behaviour and immunogenicity potential.

Caseins are a family of proteins constituted by α-caseins (αs-1 and αs-2 caseins), ß-caseins and κ-caseins. ß-caseins, in particular, show a temperature and concentration-dependent self-assembling behaviour. Recently, ß-casein micelles have been proposed as natural nanocarriers for the delivery of hydrophobic compounds, promoting their bioavailability. Until now, all studies regarding both chemical-physical characterization and applications of ß-caseins have employed the protein of bovine origin. However, it could be interesting to exploit the use of ß-caseins from other milk sources for their potential encapsulation ability and immunogenicity but, at present, no information on the self-assembling behaviour is available for ß-caseins from the milk of species different from bovine. In this work, for the first time, ß-caseins from human milk and from donkey, goat, and sheep milk were purified and their self-assembling behaviour was compared to that of a commercial bovine ß-casein, the only one for which the concentration and temperature aggregation behaviour is known. Furthermore, a preliminary evaluation of the immunogenicity potential of ß-casein from other milk sources has been performed by cross-reaction experiments using anti-ß-casein antibodies from bovine origin. The results indicated a similar self-assembling profile among all ß-caseins examined compared to the bovine ß-casein, suggesting the possible use of ß-casein from other milk sources as nanocarriers. Since donkey and human ß-casein do not cross-react with bovine anti-ß-casein antibodies, they could be particularly interesting for the development of self-assembling systems with lower hypoallergenic potential.

Comparative proteomic analysis of two clam species: Chamelea gallina and Tapes philippinarum.

Clams have long been a fisheries and aquaculture sector of great importance in Italy, the main resource of fisheries is the Chamelea gallina of indigenous origin, whereas clams breeding is supported almost entirely by the Tapes philippinarum, a species of Indo-Pacific origin. Bivalve molluscs quality depends mainly on the water quality, and then by a series of factors such as water temperature and salinity, gametogenic cycle, food availability, and environmental conditions, that affect the Condition Index. In this work crude extracts obtained from the edible part of Chamelea gallina and Tapes philippinarum were analyzed by a proteomic approach based on a two-dimensional gel electrophoresis followed by liquid chromatography-tandem mass spectrometry, in order to detect biomarkers useful for identification of the two kinds of clams and to assess their nutritional characteristics. As a result, four differentially expressed spots were found and identified, namely enolase, cyclophilin-A, ribosomal protein L13 and actin-1.

Gluten free diet and nutrient deficiencies: A review.

BACKGROUND & AIMS: The only available treatment for celiac disease (CD) is lifelong adherence to gluten free (GF)-diet. However, GF-diet may lead to possible nutrient unbalance resulting in improper nutritional quality of diet. The aim of this study is to evaluate the nutritional quality of GF-diet. METHODS: MEDLINE®/PubMed and Cochrane Library were electronically searched for articles published between 1990/01/01 and 2015/09/01. RESULTS: GF-diet was found to be poor in alimentary fiber due in particular to the necessary avoidance of several kinds of foods naturally rich in fiber (i.e. grain) and the low content of fiber of GF product that are usually made with starches and/or refined flours. Micronutrients are also found to be poor, in particular Vit. D, Vit. B12 and folate, in addition to some minerals such as iron, zinc, magnesium and calcium. Moreover, an inadequate macronutrient intake was reported related above all to the focus on the avoidance of gluten that often leaving back the importance of nutritional quality of the choice. In particular, it was found a higher content of both saturated and hydrogenated fatty acids and an increase in the glycemic index and glycemic load of the meal. CONCLUSIONS: Despite the GF-diet is necessary in celiac disease treatment and the attention is on gluten avoidance, the evaluation of nutritional quality of the diet must be considered. Moreover, educational strategies based on the relationship between nutrients and food and human health could be developed to optimize the therapeutic approach in celiac patients.

Enzymology of Pyrimidine Metabolism and Neurodegeneration.

It is well known that disorders of pyrimidine pathways may lead to neurological, hematological, immunological diseases, renal impairments, and association with malignancies. Nucleotide homeostasis depends on the three stages of pyrimidine metabolism: de novo synthesis, catabolism and recycling of these metabolites. Cytidine and uridine, in addition to be used as substrates for pyrimidine nucleotide salvaging, also act as the precursors of cytidine triphosphate used in the biosynthetic pathway of both brain's phosphatidylcholine and phosphatidylethanolamine via the Kennedy cycle. The synthesis in the brain of phosphatidylcholine and other membrane phosphatides can utilize, in addition to glucose, three compounds present in the blood stream: choline, uridine, and a polyunsaturated fatty acids like docosahexaenoic acid. Some authors, using rat models, found that oral administration of two phospholipid precursors such as uridine and omega-3 fatty acids, along with choline from the diet, can increase the amount of synaptic membrane generated by surviving striatal neurons in rats with induced Parkinson's disease. Other authors found that in hypertensive rat fed with uridine and choline, cognitive deficit resulted improved. Uridine has also been recently considered as a neuroactive molecule, because of its involvement in important neurological functions by improving memory, sleep disorders, anti-epileptic effects, as well as neuronal plasticity. Cytidine and uridine are uptaken by the brain via specific receptors and successively salvaged to the corresponding nucleotides. The present review is devoted to the enzymology of pyrimidine pathways whose importance has attracted the attention of several researchers investigating on the mechanisms underlying the physiopathology of brain.

A comparison of the carcass and meat quality of Martina Franca donkey foals aged 8 or 12 months.

The effects of slaughter age (8 vs 12 months) were investigated on meat and carcass quality obtained from Martina Franca donkey foals. Sixteen male foals were used, eight were slaughtered at 8 months of age with a mean (±s.e.) final body weight of 101±18kg and the remaining 8 foals slaughtered at 12 months of age with a mean final body weight of 122±13kg. Carcass weight and dressing percentage were higher (P<0.05) in older foals. Shear force value was lower (P<0.05) in donkeys slaughtered at 8 months of age (54.03N) compared to the same muscle Longissimus Thoracis et Lumborum (LTL) collected in older animals (62.66N). Muscle glycogen content was higher (P<0.05) in foals slaughtered at 12months of age. Donkey foal meat showed an interesting content of essential amino acids and a notable percentage of unsaturated fatty acids in both groups of animals, giving a high nutritional value to this alternative red meat.

Could albumin affect the self-assembling properties of a block co-polymer system and drug release? An in-vitro study.

PURPOSE: This work investigated the influence of a model protein, bovine serum albumin (BSA), on the properties of a thermogelling formulation intended for administration inside body compartments where there is high albumin content, as in the case of inflamed joints; it also explored the relation between the variation of these properties and release performance of methotrexate (MTX), a drug used to treat forms of arthritis and rheumatic conditions. METHODS: The influence of BSA on the micellisation and gelation behaviour of Poloxamer 407, chosen as a model copolymer, was studied by differential scanning calorimetry (microDSC), dynamic light scattering (DLS), fluorescence spectroscopy and rheology studies. A release study of MTX loaded inside the hydrogel in presence and in absence of BSA was performed. RESULTS: DLS and microDSC data revealed that the micellisation process was not affected by the protein, as demonstrated by unaltered micellar size and thermodynamic parameters. While the presence of BSA in the copolymer system reduced gel consistency, the hydrogel release performance was only slightly affected. CONCLUSION: Our results suggested that the kinetics of MTX release mainly depended on the presence of the thermogelling copolymer, although other mechanisms related to BSA could be involved. Finally, the study assessed the feasibility of using a thermogelling hydrogel for in situ drug administration in areas with the presence of high protein concentrations.

Profile of nucleosides and nucleotides in donkey's milk.

Nucleotides play a crucial role to cellular functions; they can be obtained from the diet or through the nucleotide salvage pathway, however, in particular situations (occurring mainly in newborns) the metabolic demand of nucleotides exceeds the capacity of their synthesis. These molecules, are receiving attention from a nutraceutical point of view because of their potential direct role in regulating metabolism and infant body condition. Donkey's milk may be considered a good replacer for cow's milk in feeding children with severe Ig-E mediated cow's milk protein allergy, due to its high similarity with human milk. In this study, the presence of cytidine, uridine, CMP, UMP, guanosine, and adenosine, involved in numerous biochemical and physiological activities, were detected for the first time through a RP-HPLC method.

Hypoallergenic properties of donkey's milk: a preliminary study.

Cow's 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 cow's milk substitutes. For this reason the use of milk from other mammalian species was tested. Among them, donkey's 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 donkey's milk. This study provided a preliminary evidence of the hypoallergenicity of donkey's 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.