Effects of dietary cation-anion difference on intake, milk yield, and blood components of the early lactation cow

Early lactation Holsteins cows (15 primiparous and 18 multiparous) were offered rations with dietary cation-anion difference, calculated as mEq (Na + K − Cl − S)/100 g of feed dry matter (DCAD:S), of 20, 35, or 50 mEq from d 0 (calving) to 42 d postpartum (August 20, 2000 to January 9, 2001) to determine the effects of increasing DCAD:S on dry matter intake (DMI), milk yield, and blood metabolites. For DCAD:S of 20, 35, and 50, DMI was 3.30, 3.38, 2.96 kg/100 kg of body weight (BW); milk yield was 25.5, 24.2, and 22.4 kg/d, respectively. No differences were observed for concentration or yield of milk fat or milk protein. Serum Ca, P, Mg, Na, K, Cl, cation-anion difference, insulin, and glucose did not differ with DCAD. Serum HCO₃⁻ was 26.07, 25.88, and 27.64 mEq/L for 20, 35, and 50 DCAD:S. Serum Ca, Mg, Na, and K concentrations were greater for primiparous cows (9.52 mg/dL, 2.35 mg/dL, 140.03 mEq/L, 4.66 mEq/L, respectively) than for multiparous cows (9.27 mg/dL, 2.12 mg/dL, 137.63 mEq/L, 4.46 mEq/ L, respectively). A DCAD:S between 23 and 33 mEq/ 100 g of dry matter (DM) appears to be adequate during cool weather for the milk yield that occurred in the present study based on DMI (kg/100 kg of BW), whereas DCAD:S of 50 mEq/100 g of DM may be excessive and could be too alkaline or unpalatable, resulting in decreased DMI (kg/100 kg of BW).     

Reactive intermediate in the alkali-carbonate-catalysed gasification of coal char

Based on results from a variety of experimental measurements, a detailed mechanism is postulated for the action of the inorganic catalyst in char gasification. In this mechanism, a catalyst such as potassium carbonate in contact with char undergoes a chemical and physical transformation to form a molten potassium oxide film that covers the char surface. This film serves as an oxygen transfer medium between the gaseous reactant (H₂O or CO₂) and the char. At the catalyst/char interface, an oxidation-reduction reaction occurs and the anions in the catalyst react with the oxidized char to form a phenolate-type functional group that subsequently splits out CO. The anions are replenished by reaction between the oxidizing gas (H₂O or CO₂) and the oxide at the gas/catalyst interface. Net transport of oxygen from gas to char occurs by diffusion of the species in the molten catalyst film.     

In vitro mixed ruminal microorganism fermentation of whole cottonseed coated with gelatinized corn starch and urea

We conducted an in vitro mixed ruminal microorganism fermentation study to determine the effect of coating whole cottonseed with gelatinized corn starch and feed grade urea. Treatments were arranged as a 3 x 4 factorial to provide three concentrations of starch (0.0, 2.5, and 5.0%) and four concentrations of urea (0.0, 0.25, 0.5, and 1.0%). All treatments were prepared from one lot of whole cottonseed. Batch culture fermentations were conducted using anaerobic medium that contained 20% (vol/vol) ruminal fluid in 160-ml serum bottles. Whole cottonseed was ground to pass through a 6-mm screen and weighed amounts (0, 0.4, 0.8, and 1.2 g) were added to the serum bottles. As starch increased, H2, CH4, total volatile fatty acids, and molar proportions of propionate increased linearly, whereas pH, molar proportions of acetate, and the acetate to propionate ratio decreased linearly. L-Lactate concentrations were highest with 2.5% starch compared with 0 or 5.0%. As the amount of urea in the coating increased, pH and CH4 increased linearly, whereas H2 concentrations decreased linearly. Ammonia concentrations exhibited a quadratic response due to moderate increases with the addition of 0.25 and 0.5% urea, and a greater increase was observed with 1.0% urea. Interactions between starch and urea were observed for H2, CH4, NH3, and L-lactate. Concentrations of H2 decreased and CH4 was relatively constant as urea increased in the presence of 0 and 2.5% starch, but increased with 5% starch. L-Lactate concentrations were unchanged (0% starch), higher (2.5% starch), or lower (5.0% starch) as urea increased. Ammonia concentrations increased after urea exceeded 10% of the starch concentration. The addition of urea did not prevent the decline in pH, but did reduce H2 and CH4 accumulation with 2.5% starch.     

Catechin stability of EGC- and EGCG-enriched tea drinks produced by a two-step extraction procedure

Catechin content of green tea drinks commercially available is reported to be very low in comparison with tea traditionally prepared, due to catechins conversion to their corresponding epimers during production. The purpose of this present study was to produce catechin-enriched tea drinks according to a two-step brewing procedure and to verify the catechin stability of those enriched drinks during storage. Those results confirmed that it is possible to produce EGC- and EGCG-enriched tea drinks regardless of the green tea used. Good extraction efficiencies were reached for the first and second extraction steps with catechin extraction yields ranging from 63.6% to 84.8%. Furthermore, it appeared that the catechin content in the two enriched tea drinks demonstrated a great stability since no significant degradation occurred within 8 weeks of storage. This simple two-step extraction procedure could be considered as an interesting way to produce enriched green tea drinks with more potent and stable bioactive catechins.     

Vezatin, a ubiquitous protein of adherens cell-cell junctions, is exclusively expressed in germ cells in mouse testis

In the male reproductive organs of mammals, the formation of spermatozoa takes place during two successive phases: differentiation (in the testis) and maturation (in the epididymis). The first phase, spermiogenesis, relies on a unique adherens junction, the apical ectoplasmic specialization linking the epithelial Sertoli cells to immature differentiating spermatids. Vezatin is a transmembrane protein associated with adherens junctions and the actin cytoskeleton in most epithelial cells. We report here the expression profile of vezatin during spermatogenesis. Vezatin is exclusively expressed in haploid germ cells. Immunocytochemical and ultrastructural analyses showed that vezatin intimately coincides, temporally and spatially, with acrosome formation. While vezatin is a transmembrane protein associated with adherens junctions in many epithelial cells, it is not seen at the ectoplasmic specializations, neither at the basal nor at the apical sites, in the seminiferous epithelium. In particular, vezatin does not colocalize with espin and myosin VIIa, two molecular markers of the ectoplasmic specialization. In differentiating spermatids, ultrastructural data indicate that vezatin localizes in the acrosome. In epididymal sperm, vezatin localizes also to the outer acrosomal membrane. Considering its developmental and molecular characteristics, vezatin may be involved in the assembly/stability of this spermatic membrane.     

Large-scale production of a hypoallergenic preparation of F(ab′)2 fragments from bovine colostrum

A method for the production of bovine colostral F(ab′)₂ fragments at pilot-plant scale was developed. Optimum yield of immunoglobulins in colostral whey was obtained after rennet treatment of first milking colostrum. Pepsin digestion was carried out directly on the colostral whey at pH 3·8 for 4 h, which led to the complete digestion of immunoglobulins into F(ab′)₂ fragments. Elimination of β-lactoglobulin, the main immunogenic protein, was achieved by anion exchange on Duolite A560 at pH 6·0. The preparation was diafiltered with a 5000 Da membrane and the retentate spray-dried. The powder obtained contained approximately 34% F(ab′)₂ fragments, with an antibody activity three times higher than the initial colostrum.     

Macrophage-Mediated Melanoma Reduction after HP-NAP Treatment in a Zebrafish Xenograft Model

The Helicobacter pylori Neutrophil Activating Protein (HP-NAP) is endowed with immunomodulatory properties that make it a potential candidate for anticancer therapeutic applications. By activating cytotoxic Th1 responses, HP-NAP inhibits the growth of bladder cancer and enhances the anti-tumor activity of oncolytic viruses in the treatment of metastatic breast cancer and neuroendocrine tumors. The possibility that HP-NAP exerts its anti-tumor effect also by modulating the activity of innate immune cells has not yet been explored. Taking advantage of the zebrafish model, we examined the therapeutic efficacy of HP-NAP against metastatic human melanoma, limiting the observational window to 9 days post-fertilization, well before the maturation of the adaptive immunity. Human melanoma cells were xenotransplanted into zebrafish embryos and tracked in the presence or absence of HP-NAP. The behavior and phenotype of macrophages and the impact of their drug-induced depletion were analyzed exploiting macrophage-expressed transgenes. HP-NAP administration efficiently inhibited tumor growth and metastasis and this was accompanied by strong recruitment of macrophages with a pro-inflammatory profile at the tumor site. The depletion of macrophages almost completely abrogated the ability of HP-NAP to counteract tumor growth. Our findings highlight the pivotal role of activated macrophages in counteracting melanoma growth and support the notion that HP-NAP might become a new biological therapeutic agent for the treatment of metastatic melanomas.     

Update on optimized purification and characterization of natural milk allergens

Highly purified allergens namely cow's milk alpha-lactalbumin (ALA). (Bos d 4), beta-lactoglobulin (BLG) (Bos d 5) and casein (Bos d 8) and goat's milk casein were prepared from the raw milk from a single animal with a known genetic background. Consequently the natural isoforms are limited, constant and characterized. Purification included selective precipitations and chromatographical steps. Characterization of structure and allergenic activity assessment of milk allergens were carried out using physicochemical and immunochemical methods. Taken together data demonstrated the absence of impurities and of contamination by other milk allergens in each preparation. NMR and circular dichroism analyses confirmed the native conformation and proper folding of ALA and BLG and the expected absence of folding of bovine and caprine casein. Enzyme immuno assays confirmed the native conformation of BLG and the purity and immunoreactivity of all the proteins. The allergenic activity, e. g. the IgE binding capacity, of purified proteins was identical as that of those proteins when present in milk. The purified proteins also demonstrated the ability to provoke the degranulation of humanized rat basophilic leukaemia cells. All the data thus confirm the purity, identity, structural conformation and functionality of the prepared milk allergens.