Bovine Counterpart Research Articles

Evaluation of In Vitro Antihypertensive and Anti-Inflammatory Properties of Dairy By-Products

Sweet whey (SW) and yogurt acid whey (YAW) are dairy by-products of the cheese-making process and Greek-style yogurt production, respectively. Both of them are considered pollutants with huge volumes of SW and YAW produced due to the growing demand for dairy products worldwide. Moreover, whey-derived peptides, resulting from fermentation as well as from further hydrolysis during digestion, have been associated with various biological activities. In the present study, the angiotensin-converting enzyme (ACE)-inhibitory activity of 48 SW samples and 33 YAW samples from bovine, ovine, caprine, and ovine/caprine milk obtained were evaluated. Additionally, the SW and YAW digestates and two of their fractions (smaller than 10 kDa, SW-D-P10 and YAW-D-P10, and smaller than 3 kDa, SW-D-P3 and YAW-D-P3), which were obtained after in vitro digestion and subsequent ultrafiltration, were also subjected to evaluation. Our data indicated that the D-P10 and D-P3 fractions exhibited higher ACE-inhibitory activity compared to the corresponding values before digestion. The ACE-inhibitory capacity after in vitro digestion was higher for the ovine SW samples compared to their bovine and caprine counterparts. The effect of the D-P3 fraction on the inhibition of nitric oxide (NO) production and the expression of a selected panel of immune-response-related genes in LPS-stimulated RAW 264.7 macrophages was also evaluated. Fractions from both dairy by-products inhibited NO production in LPS-stimulated RAW 264.7 cells. Especially, ovine SW-D-P3 showed a strong NO inhibitory activity and suppressed inducible nitric oxide synthase (Nos2) mRNA levels. However, YAW-D-P3 could not trigger neither the gene expression of inflammatory macrophage mediators Nos2 and cyclooxygenase-2 (Ptgs2) nor tumor necrosis factor-α (Tnf) and interleukin 6 (Il6) in LPS-stimulated murine macrophages regardless of animal origin. These findings suggest that in vitro digestion could enhance the production of ACE-inhibitory peptides in both dairy by-products, while SW from ovine origin displays higher potential as an anti-inflammatory agent, effectively preventing excessive NO production.

Effect of Milk Origin and Seasonality of Yogurt Acid Whey on Antioxidant Activity before and after In Vitro Gastrointestinal Digestion.

Yogurt acid whey (YAW) is a by-product of Greek strained yogurt production. The disposal of YAW constitutes an environmental problem, and given the increasing demand of Greek yogurt worldwide, its handling is a challenge. However, whey-derived peptides, resulting from microbial fermentation as well as those resulting from further hydrolysis during the digestion process, have been linked to enhanced biological activities. In this study, the antioxidant capacity of 33 samples of YAW obtained from Greek dairy companies of bovine, ovine or caprine origin was investigated using both cell-free and cell-based assays. The YAW samples, their in vitro digestion products (YAW-Ds) and a fraction of the digests (less than 3 kDa; YAW-D-P3) were assessed using four biochemical assays, namely ORAC, ABTS, FRAP and P-FRAP. Our data revealed a higher antioxidant capacity for digested samples compared with undigested samples, with all four methods. ORAC values after in vitro digestion were higher for the ovine samples compared to their bovine (YAW-D and YAW-D-P3) and caprine (YAW-D-P3) counterparts. Furthermore, the YAW-D-P3 fraction derived from samples collected in the summer months exhibited higher ORAC values when compared to the respective fraction from the winter months' samples. The cellular antioxidant activity of ovine YAW-D-P3 was improved in H2O2-treated HT29 cells compared to the control H2O2-treated cells. However, YAW-D-P3 could not trigger either the pathways involving the transcription factors NF-κB or NFE2L2 or the gene expression of SOD1, CAT and HMOX1 in LPS-challenged THP-1-derived macrophages. These results suggest that YAW, and particularly YAW from ovine origin, could be used as a natural source for its antioxidant potential in human and animal nutrition.

Precision-milled simulated curved root canals in bovine dentine for the assessment of chemo-mechanical root canal preparation.

The aim was to develop a standardized curved root canal model in bovine dentine and to assess whether that natural substrate would behave differently from the resin in standard plastic training blocks when prepared chemo-mechanically. The impact of substrate microhardness on simulated canal transportation was considered. High-precision computer numerical control (CNC) milling was used to recreate a simulated root canal from a resin training block (Endo Training Bloc J-Shape, size 15) in longitudinally sectioned, dis- and re-assembled bovine incisor roots. Optical overlays obtained from 10 resin blocks were used to identify an average canal and program the CNC milling apparatus accordingly. Resin and dentine microhardness were measured. Simulated root canals in resin training blocks and their bovine counterparts were then instrumented at 37°C using Reciproc R25 instruments (VDW) with water or 17% EDTA (n = 10). Open-access image processing software was used to superimpose and analyse pre- and postoperative images obtained with a digital microscope. Centering ratios were averaged to indicate canal transportation. The effects of substrate and irrigant on canal transportation were assessed by two-way anova. Superimposed images showed that resin blocks under investigation varied considerably in terms of simulated canal length and curvature, whilst the milled canals were highly similar. The microhardness of dentine was more than three times higher than that of the resin. Conversely, canal transportation was considerably greater in dentine compared to resin, and in dentine had a tendency to be increased by EDTA. There was a strong effect of substrate on canal transportation (p < .001), no overall effect of irrigant, and a marginally significant interaction between irrigant and substrate (p = .077). CNC milling allows to create standardized simulated curved root canals in bovine dentine. These models may be useful to test and compare materials and concepts of chemo-mechanical root canal instrumentation. Microhardness is a bulk feature that does not predict the response to chemo-mechanical instrumentation of a composite material such as dentine.

BAMLET (Bovine α-lactalbumin made lethal to tumor cells) inhibits autophagy flux and induces apoptosis via down-regulation of protein kinase CK1α and attenuation of the AKT/p-ß-catenin (S552) pathway in RAS-mutated human colorectal HCT 116 cells.

Oncogenic RAS mutations occur in nearly 50% of colorectal cancer cases and are usually dependent on the autophagy mechanism to maintain tumorigenesis. We have recently demonstrated that CK1α controls autophagy machinery possibly through the AKT/p-ß-catenin (S552) signaling in colorectal cancer cells harboring RAS mutation. It has been found that a lipid-protein complex comprising oleic acid binds to human α-lactalbumin, known as HAMLET (human α -lactalbumin made lethal to tumor cells), targets a broad range of kinases including CK1α. Therefore, this study was designed to investigate the effects of BAMLET (bovine α -lactalbumin made lethal to tumor cells, the bovine counterpart of HAMLET) on CK1α expression, AKT/Phospho-ß-catenin (S552) pathway, and autophagy flux in RAS-mutated human colorectal HCT 116 cells. For this purpose, HCT116 cells were treated with BAMLET and casein kinase 1 inhibitor (D4476), and quantitative real-time polymerase chain reaction (RT-qPCR) and western blot analysis were used to measure the proteins and genes of the AKT/Phospho-ß-catenin (S552) pathway and autophagy. Apoptosis was measured by flow-cytometry. We found that BAMLET significantly reduced cell viability and decreased the expression of CK1α. Additionally, BAMLET inhibited autophagy flux and enhanced the ability of CK1α inhibitor D4476 to impair autophagy flux, which was accompanied by an increase in the apoptosis percentage. We also observed that BAMLET empowered D4476 to down-regulate the AKT/Phospho-ß-catenin (S552) axis. BAMLET hampers autophagy flux and leads to apoptosis induction, possibly, by reducing the expression of CK1α and attenuation of the AKT/Phospho-ß-catenin (S552) axis.

Foaming and air-water interfacial properties of camel milk proteins compared to bovine milk proteins

The objective of this research was to explore the foaming properties of camel and bovine milk and their derived proteins fractions including sodium caseinates, sweet whey, β-casein, α-lactalbumin and β-lactoglobulin. First, camel and bovine milk proteins were identified by the reversed-phase high-performance liquid chromatography (RP-HPLC) and foaming properties (Foam capacity (FC) and stability (FS)) were analyzed. Afterwards, competitive adsorption of proteins to the air-water interface for both milk protein fractions was characterized using pendant-drop tensiometry parameters and was compared to intrinsic fluorescence results of pure proteins. Experimental results indicated that the maximum FC values were found for camel skim milk, sodium caseinates and β-casein with higher FS values for bovine β-casein. Differences in the stability and the highest tensioactive properties of camel β-casein were explained with the different molecular structure and its higher hydrophobicity when compared to its bovine counterpart. Thus, milk proteins adsorbed layers are mainly affected by the presence of β-casein which is the first adsorbed and the most abundant protein at the air-water contrary to whey proteins (α-lactalbumin and β-lactoglobulin). These globular proteins are involved in the composition of protein layers at air-water interface, giving higher viscoelastic modulus values, but could not compact well at the interface because of their rigid molecular structure. For camel milk, foaming properties and interfacial behavior are mainly maintained by camel β-casein due to its higher hydrophobicity compared to bovine β-casein and the greater exposure of tyrosine residues despite the absence of tryptophan in consistence with the intrinsic fluorescence results. Furthermore, the absence of the β-lactoglobulin leads to the dominance of the α-lactalbumin at the air-water interface which is characterized by lower hydrophobicity than its bovine counterpart leading to lower viscoelastic modulus values than those of bovine whey, and hence to weaker rheological properties of camel milk protein layer at the air-water interface.

Validation of the Bioequivalence of USP Potency Adjusted Porcine, Ovine, and Bovine Heparins

Introduction: Currently, there is a shortage of porcine heparin due to several factors such as limited availability of porcine mucosa, supply chain issues, and increased usage due to COVID-19. This has warranted the development of heparin from alternate sources such as bovine and ovine mucosa which is abundantly available for this purpose. On a mass basis, commercially available porcine heparins exhibit a similar potency (200 units/mg) to their ovine counterpart (190 units/mg) and a higher potency in contrast to their bovine counterpart (130-150 units/mg). Therefore, at gravimetric levels, the porcine heparins exhibit stronger biochemical and pharmacological effects in various laboratory assays in comparison to bovine heparin and similar effects in comparison to ovine heparins. Since heparin is standardized in biologic units and cross referenced against USP or EP Standard, it is hypothesized that potency equated porcine, ovine, and bovine heparin will exhibit similar biologic activities in laboratory assays carried out in the in vitro setting. The purpose of this study is to compare the biologic properties of the porcine, ovine, and bovine heparin at USP potency equated levels in standardized laboratory assays. Materials and Methods: Active pharmaceutical ingredients (API) of porcine mucosal heparin (200 units/mg) of U.S. origin was commercially obtained from Medefil Inc. (Glendale Heights, IL). Ovine heparin was obtained from Ronnsi Pharmaceutical (Jiangsu, China). Bovine heparin (140 units/mg) was obtained from Kin Master Pharmaceuticals (Posso Fundo, Brazil). All heparins were diluted at a concentration of 100 units/mL in saline. The anticoagulant effect of all heparins were evaluated using the whole blood clotting assays such as the ACT and thromboelastographic methods. Heparins were diluted in citrated human plasma yielding a final concentration range of 0-1 unit/mL. Clot based assays such as aPTT, TT, and prothrombinase induced clotting time (PiCT) were measured. Thrombin generation inhibition assay was carried out using a kinetic assay (CAT system, Diagnositca Stago, Paris, France). Protamine and heparinase neutralization profiles of these agents were also investigated in the plasma-based systems. These assays were then repeated at gravimetric dosages at final concentrations of 0-10 ug/mL. The results collected from these trials were then mathematically converted to units and compared to the results collected from the potency adjusted trials. All results were tabulated and compared, and applicable statistical methods were applied. Results: The USP potency adjusted heparin exhibited comparable anticoagulant effects in both the ACT and TEG assays. At equigravimetric levels porcine and ovine heparins produced comparable anticoagulant effects and bovine heparin produced weaker anticoagulant effect in both assays. In the citrated plasma supplementation studies, all drugs produced similar anticoagulant effects at potency adjusted dosages. In the chromogenic anti-Xa and anti-IIa assays, the behaviors of the agents were also comparable. In the thrombin generation assays, in terms of peak thrombin generation, area under the curve, and lag time, the porcine, ovine, and bovine heparins showed comparable effects. The protamine neutralization profiles of the porcine, ovine, and bovine heparin exhibited variable assay dependent results. Potency adjusted bovine heparin required higher amount of protamine for the complete neutralization of the biologic effects in comparison to the porcine heparin. At gravimetric concentrations, bovine heparins exhibited lower potencies than both the porcine and ovine heparins, which produced similar results. Summary and Conclusion: These results show that at potency adjusted concentrations, the porcine, ovine, and bovine heparin exhibit comparable biochemical and anticoagulant responses in the plasma-based systems. Therefore, the hypothesis that potency equated porcine, ovine, and bovine heparins exhibit comparable biochemical and anticoagulant activities is validated. Thus, the proposed approach to standardize heparins against a common standard in a biologic assay such as the USP method is valid. Furthermore, these results warrant regulatory considerations to fast track the review process for the re-introduction of bovine heparin and approval of bovine heparin as a biosimilar anticoagulant to porcine heparin. Disclosures No relevant conflicts of interest to declare.

Potency Equated Porcine and Bovine Mucosal Heparin Are Bioequivalent in Terms of Biochemical and Pharmacological Effects

Introduction: Currently, there is a shortage of porcine heparin due to limited availability of porcine mucosa and supply chain issues. Bovine heparin has been used for clinical purposes globally and is being considered for reintroduction in the U.S. On a mass basis, commercially available porcine heparins exhibit a higher potency (180-220 units/mg) than their bovine counterpart (130-150 units/mg). Therefore, at gravimetric levels, the porcine heparins exhibit stronger biochemical and pharmacological effects in comparison to bovine heparin. Since heparin is standardized in biologic units relative to the USP or EP Standard, it is hypothesized that potency equated porcine and bovine heparin will exhibit similar biologic activities in laboratory assays. The purpose of this study was to compare the biologic properties of the porcine and bovine heparin at USP potency equated levels in standardized laboratory assays used for measuring and monitoring heparins. Materials and Methods: Active pharmaceutical ingredient (API) porcine mucosal heparin (200 units/mg) of U.S. origin was obtained from Medefil Inc. (Glendale Heights, IL). Bovine heparin (140 units/mg) was obtained from KinMaster Produtos Químicos (Passo Fundo, Brazil). Whole blood Activated Clotting Time (ACT) and Thrombelastographic analyses were performed. Clot based assays such as aPTT, TT, and prothrombinase induced clotting time (PiCT) were performed using citrated human plasma supplemented with heparins up to 1 anti-Xa unit/mL. Anti-Xa and anti-IIa activities were determined using chromogenic antiprotease assays (Biophen Kits). In a purified antithrombin-supplemented system, the inhibitory effects of these agents were measured in terms of anti-Xa and anti-IIa activities and expressed as IC50 values. Thrombin generation inhibition was measured using a kinetic assay (CAT system, Diagnostica Stago, Paris, France). Protamine and platelet factor 4 neutralization profiles were investigated in plasma-based systems. The heparin induced thrombocytopenia antibody (HIT) interaction profile was assessed using platelet aggregometry. Results: Both heparins produced comparable prolongation of the ACT to a range of 200-250 seconds. The thrombelastographic profile was comparable at concentrations of 0.125 unit/mL and 2.5 units/mL. Whole blood and plasma clotting times (aPTT and TT assays) were comparable at concentrations up to 1 unit/mL. In the chromogenic anti-Xa and anti-IIa assays, the behavior of both agents was also comparable. In the purified system, the IC50 values for both agents for the anti-Xa activity ranged from 0.33-0.40 unit/mL, whereas the anti-IIa values ranged from 0.38-0.42 unit/mL; no significant differences were noted between the porcine and bovine heparins. In the thrombin generation assays, in terms of peak thrombin generation, area under the curve, and lag time, both the porcine and bovine heparins showed comparable effects. The protamine neutralization profiles of the porcine and bovine heparin exhibited variable assay dependent results. Potency adjusted bovine heparin required higher amount of protamine for the complete neutralization of the biologic effects in comparison to the porcine heparin. Similar results were obtained in the platelet factor 4 neutralization studies. In the functional HIT screening assay, both heparins exhibited a similar concentration dependent aggregation of human platelets. These results are summarized in the Table. Summary and Conclusion: These results show that at potency adjusted concentrations, porcine and bovine heparin exhibit comparable biochemical and anticoagulant responses in whole blood, plasma based, and purified biochemical assays. Most notably, both the porcine and bovine heparin produced comparable HIT antibody mediated aggregation responses. Thus, the proposed approach to standardize heparins against a common standard using a biologic assay such as the USP method is valid. These results warrant additional validation studies to reintroduce bovine heparin for clinical use. Table Disclosures Jeske: KinMaster Produtos Quimicas: Research Funding.

Comparison of human and bovine dental enamel by TEM and t-EBSD investigations

The microstructures of human dental enamel and bovine enamel are compared to each other. To obtain samples for transmission electron microscopy (TEM), focused ion-beam (FIB) milling is used. The preparation of such TEM-slices is found to be very effective when operating the FIB with adapted parameters. After the milling process, the TEM-slices are then thinned by means of the ion beam to achieve samples being transparent for the electron beam. With a home-built sample holder, the electron backscatter diffraction (EBSD) can be operated in the scanning electron microscope (SEM) in transmission mode called t-EBSD. This technique enables the crystallographic orientation measurement on nanometer-sized, non-conducting enamel grains with a reasonable quality. Both TEM and t-EBSD images reveal a similar arrangement of the apatite crystals within the enamel, but it is obvious that the nanostructure of human enamel follows a more complex construction principle. The grain sizes of bovine enamel are much larger, and it is difficult to recognize the chain arrangement as found previously in the human enamel. As a result of the comparison, one can state that the nanostructure of human enamel is clearly more complicated than the bovine counterpart.

Effect of nonenzymatic deamidation on the structure stability of Camelus dromedarius α-lactalbumin

Camelid α-lactalbumin is the only known protein that can undergo nonenzymatic deamidation on two Asn residues. This leads to the generation of a mixture of unusual isoAsp and d-Asp residues that may impact health. The effect of deamidation on camel α-lactalbumin instability was investigated. Circular dichroism showed that the altered protein acquired secondary structure resulting in an increase in α-helix content. In good agreement, the 3D structure of camel α-lactalbumin determined by X-ray crystallography, displayed a short additional α-helix probably induced by deamidation, compared to the human and bovine counterparts. This α-helix was located in the C-terminal region and included residues 101–106. Differential scanning calorimetry together with the susceptibility to thermolysin showed that the deamidation process reinforced the structural stability of the α-lactalbumin at high temperature and its resistance toward proteolysis.

USP Potency Equated Enriched Bovine and Porcine Mucosal Heparins Exhibit Comparable Biochemical and Pharmacological Effects

ObjectiveCommercially available native porcine (NP) heparins exhibit a higher potency (200 U/mg) in contrast to their native bovine (NB) counterpart (140 U/mg). Enriched versions of porcine heparins (EP: 240 U/mg) and bovine heparins (EB: 180 U/mg) were developed using improved separation methods. At a gravimetric concentration, porcine heparins (NP and EP) exhibit stronger effects in contrast to bovine heparins. The purpose of this study is to compare the biologic properties of these agents at equated USP potencies.MethodsNP, EP, NB, and EB heparins were obtained from Yino Pharma (China). A commercially available porcine mucosal heparin was obtained from Hospira/Pfizer (USA) (190 U/mg). Working dilutions were prepared at potency equated levels of 10 U/mL. Whole blood aPTT and TT were measured on the ST4 instrument at a concentration range of 0–1 U/mL. Whole blood ACT was measured using a Hemochron instrument at a fixed concentration of 1 U/mL. TT, aPTT, and protamine neutralization profiles were also measured in plasma supplemented with each agent at a concentration range of 0–1 U/mL. The USP potency assay was carried out using chromogenic antiprotease assays (Biophen Kits). The inhibitory concentrations at 50% (IC‐50) were measured using a purified antithrombin biochemical system employing a kinetic method. Thrombin generation inhibition assay was carried out using a CAT method (Diagnositca Stago, Paris, France).ResultsIn the whole blood ACT method, all drugs produced comparable effects with values between 200–250 seconds. In the whole blood aPTT and TT assays, all results were comparable. In the plasma supplementation studies, all drugs were comparable in the aPTT, factor Xa, and factor IIa studies and the protamine neutralization profiles. However, in the TT assay, NB and EP were somewhat weaker. The anti‐Xa and anti‐IIa activities of all drugs were comparable. The IC‐50 values for the anti‐Xa ranged from 0.33–0.40 u/mL and the anti‐IIa values ranged from 0.38–0.42 U/mL. In the thrombin generation assays, the EB heparins showed slightly weaker activities.ConclusionThese results show that at potency equated concentrations, both the native and enriched porcine and bovine heparin preparations exhibit comparable biochemical and anticoagulant responses. Therefore, the hypothesis that potency equated native/enriched bovine and porcine heparins exhibit comparable biochemical and anticoagulant activities is validated.Support or Funding InformationNoneThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Characterization of the complex between native and reduced bovine serum albumin with aquacobalamin and evidence of dual tetrapyrrole binding.

Serum albumin binds to a variety of endogenous ligands and drugs. Human serum albumin (HSA) binds to heme via hydrophobic interactions and axial coordination of the iron center by protein residue Tyr161. Human serum albumin binds to another tetrapyrrole, cobalamin (Cbl), but the structural and functional properties of this complex are poorly understood. Herein, we investigate the reaction between aquacobalamin (H2OCbl) and bovine serum albumin (BSA, the bovine counterpart of HSA) using Ultraviolet-Visible and fluorescent spectroscopy, and electron paramagnetic resonance. The reaction between H2OCbl and BSA led to the formation of a BSA-Cbl(III) complex consistent with N-axial ligation (amino). Prior tothe formation of this complex, the reactants participate in an additional binding event that has been examined by fluorescence spectroscopy. Binding of BSA to Cbl(III) reduced complex formation between the bound cobalamin and free cyanide to form cyanocobalamin (CNCbl), suggesting that the β-axial position of the cobalamin may be occupied by an amino acid residue from the protein. Reaction of BSA containing reduced disulfide bonds with H2OCbl produces cob(II)alamin and disulfide with intermediate formation of thiolate Cbl(III)-BSA complex and its decomposition. Finally, in vitro studies showed that cobalamin binds to BSA only in the presence of an excess of protein, which is in contrast to heme binding to BSA that involves a 1:1 stoichiometry. In vitro formation of BSA-Cbl(III) complex does not preclude subsequent heme binding, which occurs without displacement of H2OCbl bound to BSA. These data suggest that the two tetrapyrroles interact with BSA in different binding pockets.

A 100-Year Review: A century of change in temperate grazing dairy systems

From 1917 to 2017, dairy grazing systems have evolved from uncontrolled grazing of unimproved pastures by dual-purpose dairy-beef breeds to an intensive system with a high output per unit of land from a fit-for-purpose cow. The end of World War I signaled significant government investments in agricultural research institutes around the world, which coincided with technological breakthroughs in milk harvesting and a recognition that important traits in both plants and animals could be improved upon relatively rapidly through genetic selection. Uptake of milk recording and herd testing increased rapidly through the 1920s, as did the recognition that pastures that were rested in between grazing events yielded more in a year than those continuously grazed. This, and the invention and refinement of the electric fence, led to the development of "controlled" rotational grazing. This, in itself, facilitated greater stocking rates and a 5 to 10% increase in milk output per hectare but, perhaps more importantly, it allowed a more efficient use of nitrogen fertilizer, further increasing milk output/land area by 20%. Farmer inventions led to the development of the herringbone and rotary milking parlors, which, along with the "unshortable" electric fence and technological breakthroughs in sperm dilution rates, allowed further dairy farm expansion. Simple but effective technological breakthroughs in reproduction ensured that cows were identified in estrus early (a key factor in maintaining the seasonality of milk production) and enabled researchers to quantify the anestrus problem in grazing herds. Genetic improvement of pasture species has lagged its bovine counterpart, but recent developments in multi-trait indices as well as investment in genetic technologies should significantly increase potential milk production per hectare. Decades of research on the use of feeds other than pasture (i.e., supplementary feeds) have provided consistent milk production responses when the reduction in pasture intake associated with the provision of supplementary feed (i.e., substitution rate) is accounted for. A unique feature of grazing systems research over the last 70 yr has been the use of multi-year farm systems experimentation. These studies have allowed the evaluation of strategic changes to a component of the system on all the interacting features of the system. This technique has allowed excellent component research to be "systemized" and is an essential part of the development of the intensive grazing production system that exists today. Future challenges include the provision of skilled labor or specifically designed automation to optimize farm management and both environmental sustainability and animal welfare concerns, particularly relating to the concentration of nitrogen in each urine patch and the associated risk of nitrate leaching, as well as concerns regarding exposure of animals to harsh climatic conditions. These combined challenges could affect farmers' "social license" to farm in the future.

Characteristics and Gel Properties of Gelatin from Goat Skin as Affected by Extraction Conditions

Characteristics and gel properties of gelatin from goat skin, as influenced by extraction conditions, were studied. Yield of gelatin was 22.1–23.1% (wet weight basis). All gelatins contained α-chains as the predominant components, followed by β-chain. However, no β-chain was retained when extracted at 70C. Gelatin extracted at 50C for 2.5 h exhibited the highest gel strength (267 g) (P < 0.05). Gelling and melting temperatures for goat skin gelatin were in the ranges of 21.18–25.17C and 30.69–34.12C, respectively. Gel strength, gelling and melting temperatures of gelatin gels generally decreased with increasing extraction temperatures and times. Gels of gelatin extracted at a higher temperature for longer time had larger strands and voids. Gelatin extracted at 50C for 2.5 h contained imino acids of 225 residues/1,000 residues and showed higher gel strength than bovine gelatin. Thus, it could be used as a potential replacer for commercial gelatins. Practical Appications Gelatin has been used in food and non-food industries all over the world. With increasing demand of gelatin, particularly as the halal ingredient, the promising alternative raw material has been searched. Goat skin has become the potential collagenous starting material; however, the extraction conditions must be optimized to obtain the good quality gelatin. Therefore, gelatin from goat skin with properties equivalent to commercially available bovine or porcine counterparts can be produced and the goat skin can be better exploited.

Bioactive Proteins in Human Milk: Health, Nutrition, and Implications for Infant Formulas

Breast milk confers many benefits to the newborn and developing infant. There is substantial support for better long-term outcomes, such as less obesity, diabetes, and cardiovascular disease, in breastfed compared with formula-fed infants. More short-term outcomes, such as incidence and duration of illness, nutrient status, and cognitive development during the first year of life also demonstrate benefits of breastfeeding. Several proteins in breast milk, including lactoferrin, α-lactalbumin, milk fat globule membrane proteins, and osteopontin, have been shown to have bioactivities that range from involvement in the protection against infection to the acquisition of nutrients from breast milk. In some cases, bovine counterparts of these proteins exert similar bioactivities. It is possible by dairy technology to add protein fractions highly enriched in these proteins to infant formula.

Angiotensin I-converting (ACE)-inhibitory and anti-inflammatory properties of commercially available Greek yoghurt made from bovine or ovine milk: A comparative study

The objective of this study was to compare the presence of peptides with angiotensin I-converting (ACE)-inhibitory and immunomodulatory activities in commercial Greek yoghurt made from bovine or ovine milk. Water soluble extracts (WSE) in yoghurts made from ovine milk exhibited higher ACE-inhibitory activity than their bovine counterparts. Similarly, ovine yoghurt WSE inhibited more effectively the expression of two pro-inflammatory genes (inducible NO synthase and cyclooxygenase-2) by ovine monocytes. Following gel filtration of WSE by Sephadex G25, one fraction exhibited only ACE-inhibitory activity, two only immune inhibitory while one possessed both activities. Thus, selected peptidic fractions mimicked the original effects observed in WSE prior to fractionation.

Cholesterol Concentrations and Lipolytic Characteristics of Commercial Bovine and Caprine Milk Yogurts during Four Weeks Refrigerated Storage

Yogurt is a popular cultured dairy product in many countries, while few studies have been reported on cholesterol concentration and lipolytic property of goat milk yogurt. The purposes of this study were to quantify cholesterol contents of commercial bovine and caprine milk yogurts produced in the US, and compare differences in lipolysis between the two species products during 0, 2 and 4 weeks storage at 4oC. Three batches each of commercial cow milk yogurt (CCY) and commercial goat milk yogurt (CGY) were purchased from local retail stores. All experimental yogurt samples were subjected to analyses of cholesterol, basic nutrients, pH and acid degree value (ADV) using AOAC procedures. Cholesterol content was quantified by GC-17A (Shimadzu Scientific, Inc., Columbia, MD) equipped with a fused silica capillary column (HP-5, 30 m × 0.32 mm inner diameter × 0.25 μm thickness) and FID detector. The respective mean (%) of protein, fat, ash, carbohydrates and total solids of the CGY and CCY were: 3.25, 3.69; 2.97, 2.83; 0.847, 0.777; 4.19, 4.89; 11.95, 11.49, indicating that the CGY contained higher fat, ash and total solids, and lower protein than those of the CCY. The higher carbohydrates, ash and total solids in the CGY accounted for the supplementation of tapioca and pectin in the caprine product. Cholesterol contents (mg/100 g) of the CGY and CCY for 0, 2 and 4 weeks storage were: 19.8, 18.4; 20.3, 17.9; 22.0, 19.7, respectively, showing that CGY contained slightly higher cholesterol than CCY throughout the storage periods. It was concluded that the caprine yogurt contained slightly higher fat and cholesterol than the bovine counterpart, which was unexpected because goat milk is known to contain lower cholesterol than cow milk. The differences in lipolysis between CGY and CCY during the 4 weeks storage were minimal.

Molecular characterization and in-silico analysis of complete coding sequence of bubaline mLYS

The role of immune-relevant genes involved in disease biology as well as innate and adaptive immunity is at the centre stage while exploring the genetic basis of disease occurrence. The macrophage expressed lysozyme gene product contributes to innate immunity by cleaving the ß- 1,4 linkage of bacterial peptidoglycan in phagocytosis. Higher specific activity observed in bubaline milk/ serum lysozymes over their bovine counterparts is expected to contribute higher resistances to diseases in general and mastitis in particular. To explore the reasons for higher activity in terms of sequence variations, macrophage expressed lysozyme cDNA was synthesized, cloned and sequenced. The 593 bp mRNA sequence revealed a 444 bp (27nt to 470nt) ORF bearing usual start codon ATG and end codon TAA with GC content of 46 % coding for precursor polypeptide of 147 amino acids. The comparative sequence analysis of cattle and buffalo revealed the difference at 10 places leading to 3 non-synonymous (5th, 116th and 142th) substitutions which did not affect the predicted 3D structure. The mLYS polypeptide had signal sequence (1st to 18th residue) and the mature peptide had two lysozyme catalytic sites, three Ca++ binding sites as well as eleven catalytic clefts prominent among the conserved domains. Secondly, bubaline mLYS was phylogenetically closer to the abomasum type than mammary gland type. The high similarity in the coding sequences and predicted structure suggested that bubaline lysozyme gene if hyper-expressed either in native or recombinant form in bovine udder may result in better udder health.

Comparison of dipeptidyl peptidase IV-inhibitory activity of peptides from bovine and caprine milk casein by in silico and in vitro analyses

The different potential for release of dipeptidyl peptidase IV (DPP-IV)-inhibitory peptides from bovine and caprine milk casein was assessed by in silico analysis and in vitro proteolytic assays. The former predicted a weakly higher potency for caprine casein than bovine as a precursor of DPP-IV-inhibitory peptides, although with markedly diverse potential for individual caseins. This was verified by protease hydrolysis; trypsin-treated casein hydrolysates (TCAHs) displayed the strongest bioactivity. Fractionation of caprine TCAHs revealed slightly, but significantly, higher inhibitory activity in peptides <5 kDa, and notably greater efficiency in the >5 kDa fraction, than their bovine counterparts. Through in silico trypsin hydrolysis and peptides synthesis, four novel caprine casein-derived DPP-IV-inhibitory peptides, including GPFPILV and HPINHR (half maximal inhibitory concentration = 163.7 ± 1.33 and 452.2 ± 7.15 μm, respectively), were found. This study corroborates the weak superiority of caprine casein over bovine in release of DPP-IV-inhibitory peptides, and identifies several novel caprine casein-derived DPP-IV-inhibitory peptides.

Structural basis of glycan specificity in neonate-specific bovine-human reassortant rotavirus.

Strain-dependent variation of glycan recognition during initial cell attachment of viruses is a critical determinant of host specificity, tissue-tropism and zoonosis. Rotaviruses (RVs), which cause life-threatening gastroenteritis in infants and children, display significant genotype-dependent variations in glycan recognition resulting from sequence alterations in the VP8* domain of the spike protein VP4. The structural basis of this genotype-dependent glycan specificity, particularly in human RVs, remains poorly understood. Here, from crystallographic studies, we show how genotypic variations configure a novel binding site in the VP8* of a neonate-specific bovine-human reassortant to uniquely recognize either type I or type II precursor glycans, and to restrict type II glycan binding in the bovine counterpart. Such a distinct glycan-binding site that allows differential recognition of the precursor glycans, which are developmentally regulated in the neonate gut and abundant in bovine and human milk provides a basis for age-restricted tropism and zoonotic transmission of G10P[11] rotaviruses.

Anti-inflammatory and Anti-Allergic Properties of Donkey’s and Goat’s Milk

Nowadays, donkey's and goat's milk consumption has been reevaluated for its potential benefits to human health. For example, in infants with intolerance to cow's milk, donkey's milk represents a good alternative due to its chemical characteristics similar to those of human milk. On the other hand, goat's milk in virtue of its higher content in short chain, medium chain, mono and polyunsaturated fatty acids than that of cow's milk, is more digestible than the bovine counterpart. From an immunological point of view, donkey's milk is able to induce release of inflammatory and anti-inflammatory cytokines from normal human peripheral blood lymphomononuclear cells, thus maintaining a condition of immune homeostasis. Similarly, goat's milk has been shown to trigger innate and adaptive immune responses in an in vitro human system, also inhibiting the endotoxin-induced activation of monocytes. Finally, in these milks the presence of their own microbiota may normalize the human intestinal microbiota with a cascade of protective effects at intestinal mucosal sites, even including triggering of intestinal T regulatory cells. In the light of the above considerations, donkey's and goat's milk should be recommended as a dietary supplement in individuals with inflammatory and allergic conditions, even including elderly people.