To maintain activity in a coenzyme/enzyme mixture system, such as ${\beta}$-nicotinamide adenine dinucleotide (NADH)/dehydrogenase, the water-soluble 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers as an additive were synthesized and investigated for their stabilizing function. The inhibitor for the NADH/dehydrogenase reaction was spontaneously formed when the NADH was stored in the dehydrogenase solution. Therefore, we hypothesized that if the additive polymer could interact with an inhibitor without any adverse effect on the dehydrogenase, the activity in the NADH/dehydrogenase mixture could be maintained. We selected lactose dehydrogenase (LDH) as the enzyme, and the NADH was dissolved and incubated at $37^{\circ}C$ in the LDH solution containing the polymers. The phospholipid polymers used in this study were poly(MPC) (PMPC), poly(MPC-co-3-trimethylammonium-2-hydroxypropyl methacrylate chloride) (PMQ) and poly[MPC-co-potassium 3-methacryloyloxypropyl sulfonate ($MSO_3$)] ($PMMSO_3$). The poly($MSO_3$) was used as a reference. For the PMQ and $PMSO_3$ aqueous solutions, the activity of the NADH/LDH mixture system decreased with incubation time as the same level or lower than that in the Tris buffered solution in the absence of the polymers. However, for the poly($MPC-co-MSO_3$) ($PMMSO_3$) aqueous solution, the activity of the NADH/LDH mixed system was six times higher than that in the buffered solution even after a 3-days incubation. The LDH activity was 1.5-1.8 times higher in the presence of the $PMMSO_3$ compared with that in the $PMSO_3$ solution. The mixture of two polymers, poly(MPC) and poly($MSO_3$), did not produce any stabilization. Thus, both the MPC and $MSO_3$ units in the polymer chain had important and cooperative effects for stabilizing the NADH/LDH mixture.
Kolenda, Magdalena;Sitkowska, Beata;Kamola, Dariusz;Lambert, Barry D.
Animal Bioscience
/
v.34
no.8
/
pp.1283-1289
/
2021
Objective: The progestogen-associated endometrial protein (PAEP) gene encodes the main whey protein in milk, β-lactoglobulin. The aim of the study was to investigate polymorphism in the PAEP gene and its association with milk yield, composition, and quality. Methods: Test-day records for 782 dairy cows were analysed. A total of 10 single nucleotide polymorphisms (SNP) within the PAEP gene were investigated. The following parameters were recorded: milk yield (MY, kg/d), percent milk fat (%), protein (PP, %), dry matter (DMP, %) and lactose (LP, %), urea content (UC, mg/L) as well as natural logarithm for somatic cell count (LnSCC, ln). Effect on genomic estimated breeding values accuracy was evaluated with pedigree and single step model. Results: Results show that only three SNPs were polymorphic, creating 5 composite genotypes: P1 to P5. Differences in MY between composite genotypes were noted in the two tested herds. Cows with P5 composite genotypes were characterised by the highest PP and LnSCC and the lowest LP and UC (p<0.05). P4 was linked to an increased DMP and UC, while P3 to an increase in LP and decrease in PP and LnSCC. Both factors are important markers in herd management and have high influences on the herds economics. For 5 out of 7 traits the accuracy of prediction was improved by including the haplotype as a fixed effect. Conclusion: Presented results may suggest a new way to optimise breeding programmes and demonstrate the impact of using genomic data during that process.
In this study, physicochemical properties and the antioxidative activity of whey protein isolate(WPI) for com germ oil were measured. The pH of WPI was 6.26, and the titrable acidity was 0.18%. The WPI’s moisture content was 5.2% and each of the other element content such as lactose, crude protein, crude ash and crude fat was found to be 0.8%, 90.7%, 2.7% and 0.6%, respectively. The amounts of active SH group in WPI 9 ${\mu}$ M-g and total colony counts of bacteria was 5.9 ${\times}$ 10$^3$ CFU-g. ${\alpha}$-Lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin(BSA) were shown in WPI as major protein by electrophoresis. The antioxidative effect of WPI and other antioxidants on com germ oil used as substrate was determined by peroxide value(POV) and conjuqated dienoic acid value(CDV). By these results, the order of antioxidative effects could be defined as BHT 0.02%>ascorbic acid 0.1%>WPI 0.1%>WPI 0.02%>ascorbic acid 0.02%>control>tocopherol 0.02%>tocopherol 0.1%, respectively. Also the induction period of com germ oil added with WPI was longer by 1.6 times than that of control(none added any antioxidant). Therefore the fact suggested that WPI could be utilized as a good antioxidative agents.
Heo, Young Tae;Ha, Woo Tae;Lee, Ran;Lee, Won-Young;Jeong, Ha Yeon;Hwang, Kyu Chan;Song, Hyuk
Asian-Australasian Journal of Animal Sciences
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v.30
no.6
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pp.878-885
/
2017
Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.
Chaiyabutr, Narongsak;Thammacharoen, S.;Komolvanich, S.;Chanpongsang, S.
Asian-Australasian Journal of Animal Sciences
/
v.20
no.9
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pp.1407-1416
/
2007
Ten, first lactation, 87.5%HF dairy cattle were used to investigate effects of long-term administration of recombinant bovine somatotropin (rbST) on nutrient uptake by the mammary gland at different stages of lactation. Measurements of arterial plasma concentrations and arterial-venous differences of metabolites across the mammary gland were performed in combination with measurment of mammary blood flow to estimate the mammary uptake. Animals in experimental groups were injected subcutaneously every 14 days from day 60 of lactation with a prolonged-release formulation of 500 mg of rbST (POSILAC, Monsanto, USA) or with sterile sesame oil without rbST in the control group. During early lactation, the milk yield of rbST-treated animals was higher than that of the control animals (p<0.05). The peak milk yield in both groups of animals declined from the early period of lactation with progression to mid- and late-lactation. No significant changes were observed in the concentration of milk lactose, while the concentrations of milk protein significantly increased as lactation advanced to mid- and late-lactation in both groups. Milk fat concentrations were significantly higher in rbST-treated animals than in control animals, particularly in early lactation (p<0.05). Mammary blood flow (MBF) markedly increased during rbST administration and was maintained at a high level throughout lactation. The mean arterial plasma concentrations for glucose and acetate of rbST-treated animals were unchanged. The net mammary glucose uptake of rbST-treated animals increased approximately 20% during early lactation, while it significantly decreased (p<0.05), including the arteriovenous differences (A-V differences) and extraction ratio across the mammary gland, as lactation advanced to mid- and late-lactation. A-V differences, mammary extraction and mammary uptake for acetate increased during rbST administration and were significantly higher (p<0.05) than in the control animals in early and mid-lactation. Mean arterial plasma concentrations for ${\beta}$-hydroxybutyrate and free glycerol were unchanged throughout the experimental periods in both groups. A-V differences and extraction ratio of ${\beta}$-hydroxybutyrate across the mammary gland did not alter during rbST administration. Mean arterial plasma concentrations for free fatty acids ($C_{16}$ to $C_{18}$), but not for triacylglycerol, increased in rbST-treated animals and were significantly higher than in control animals during early lactation (p<0.01). These findings suggest that an increase in MBF during rbST administration would not be a major determinant in the mediation of nutrient delivery and uptake by the mammary gland for increased milk production. Local changes in biosynthetic capacity within the mammary gland would be a factor in the utilization of substrates resulting in the rate of decline in milk yield with advancing lactation.
Journal of the Korean Society of Food Science and Nutrition
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v.22
no.2
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pp.208-214
/
1993
Ginseng-whey beverages were prepared with rennet whey, ginseng, sweetener, honey and Japanese apricot, inoculated with different strains of lactic acid bacteria or unfermented partly. The samples were stored at 4$^{\circ}C$ or 30$\pm$1$0^{\circ}C$ and then physicochemical and microbiological properties were investigated. The yield of whey was 78.8%. The pH-values reduced and acidities increased during the storage period. The contents of solid-substances, ash and lipid in ginseng-whey beverages were 7.90~8.20%, 0.62~0.66% and 0.16%, respectively. The protein contents of ginseng-whey beverages were 0.42~0.56% and the contents were not changed during the storage period. The lactose contents of fermented beverages were higher than those of unfermented beverages. During the storage period (1~5 weeks), the ranges of D(-) - and L(+)- lactic acid contents in fermented ginseng-whey beverages (17.3~156.1 mg/100g, 347.3~1894.2mg/100g) were higher than those of unfermented ginseng-whey beverages (6.2~82.8mg/100g, 7.1~885.5mg/100g). The contents of total saponin in unfermented sample and fermented sample (Lac. casei sub-sp. casei+Str. salivarius sub-sp. thermophilus) were increased during the storage period. But, those of the fermented sample(Lac. acidophilus+Lac. delbrueckii sub-sp. bulgaricus) were reduced. In the electrophoretic results of ginseng-whey beverages, an $\alpha$-lactalbumin and a $\beta$-lactoglobulin bands were shown apparently and there were no changes observed during the storage period. During the storage period (1~3 week) the coliform was not detected and total plate counts and psychrotrophs were increased according to the storage period.
This experiment was carried out to examine the fermentation properties of yogurt with Lycii fructus, Lycii folium and Lycii cortex powder, and extract additives at concentrations of 0.5, 1.0, 2.0, 4.0, and 6.0%. Lactic acid bacteria was used in a mixed starter culture of Streptococcus salivarius ssp. thermophilus(ST36) and Lactobacillus delbrueckii ssp. bulgaricus(LB12). When the boxthorn was added with extract types, the changes of pH, acidity and lactic acid bacteria counts of yogurt during the fermenation of 3 hours were pH 5.64, titratable acidity 0.85%, 5.80xl0$\^$6/cfu/ml of viable cell counts for control yogurt, whereas those were pH 4.10∼5.06, titratable acidity 0.98∼1.27%, 1.80∼9.60x10$\^$7/ cfu/ml of viable cell counts for Lycii fructus extract yogurt. The lactose hydrolysis ratio was better for 1.0% Lycii fructus extract yogurt(42.00%) and 1.0% Lycii folium extract yogurt(41.46%) than for control yogurt(28.40%). Also, content of lactic acid of 1.0% Lycii fructus(11.9 times) and 1.0% Lycii folium extract yogurt(10.6 times) produced more than control yogurt(7.3 times). The viscosity of yogurt was better for boxthorn extract yogurt(1,027∼1,382 cps) than for control yogurt(975cps). The sensory scores of color, taste and overall acceptability of yogurt with 0.5, and 1.0% Lycii fructus extract additive were better than other groups. The yogurts made with increased Lycii fructus extract concentration(0.5∼6.0%), showed the increase of lactic acid, titratable acidity, number of lactic acid bacteria, viscosity and lactose hydrolysis rate compared to the treatments of 0.5, 1.0, 2.0, and 4.0% Lycii folium and Lycii cortex extract and powder yogurt. We gained excellent results from the yogurt to which Lycii fructus extract was added with 0.51.0% concentration.
The purpose of this study was to improve the production of Mozzarella cheese analogues manufactured using mixtures of soy milk and concentrated raw milk by performing ultrafiltration (UF) and to assess the quality of these cheeses during a 30-day storage period at $4^{\circ}C$, relative to that of Mozzarella cheese manufactured with the traditional method. The solid consistency of Mozzarella cheese analogue prepared from milk mixtures was lower than that of cheese manufactured from raw milk or soy milk and increased during storage, which is considered to be the result of decreasing water levels, as well as with increasing soy milk concentrations. In the Mozzarella cheese analogue generated using the milk mixtures, the fat content decreased with increase in the soy milk concentration, while it decreased during the storage period. Lactose levels were lowest in cheese composed of soy milk or raw milk and processed by UF, and decreased during storage in cheese produced using milk mixtures. In milk mixtures containing soy milk, the protein concentration increased with increasing amounts of raw milk and did not change during the storage period. The water-soluble nitrogen compound level was similar between cheeses and increased only slightly during storage. The amount of non-protein nitrogen compounds was higher in the cheese analogue than in the control cheese and tended to increase during storage. Analysis of the physicochemical traits of the Mozzarella cheese analogue yielded the following results: During storage, titratable acidity levels increased while pH tended to decrease. After analysis using electropherograms, it was classified as ${\alpha}$-, ${\beta}$-, or ${\kappa}$-casein. The results of rheometry tests showed that in the Mozzarella cheese analogue prepared from milk mixtures, with raw milk concentrated by UF, increases in concentration rate lead to lowered hardness, elasticity, cohesiveness, and brittleness. When cheese was produced from milk mixtures and concentrated by UF, meltability increased as the concentration rate increased, although to an extent that was less than that observed for the control cheese, and tended to increase during storage. Sensory evaluation showed that the analogue cheese was much better than the control cheese in terms of formation, appearance, and flavor.
Solid dispersions were prepared to increase the dissolution rate of biphenyl dimethyl dicarboxylate (DDB) using water-soluble carriers such as povidone, copolyvidone, $2-hydroxypropyl-{\beta}-cyclodextrin (HPCD)$, sodium salicylate or sodium benzoate by solvent evaporation method. Solid dispersions were characterized by infrared spectrometry, differential scanning calorimetry (DSC) and powder X-ray diffractometry, dissolution and permeation studies. DDB tablets (7.5 mg) were prepared by compressing the powder mixtures composed of solid dispersions, lactose, com starch, crospovidone and magnesium stearate using a single-punch press. DDB capsules (7.5 mg) were also prepared by filling the mixtures in empty hard gelatin capsules (size No.1). From the DSC and powder x-ray diffractometric studies, it was found that DDB was amorphous in the HPCD or copolyvidone solid dispersions. Dissolution rates after 10 min of DDB alone and solid dispersions (1 : 10) in sodium benzoate, sodium salicylate and copolyvidone were 11.8, 23.5, 22.8 and 82.5%, respectively. Dissolution rates of DDB after 30 min from 1 : 10 and 1 : 20 copolyvidone solid dispersions were 80.5 and 95.0%, respectively. For the DDB tablets prepared using solid dispersions (1 : 20), the initial dissolution rate was dependent on carrier material, and was ranked in order, $Kollidon\;30\;{\ll}$ copolyvidone < HPCD. For the HPCD solid dispersion tablets, dissolution rate reached 97.4% after 15 min, but thereafter slowly decreased to 80.7% after 2 hr due to the precipitation of DDB. However, in the case of copolyvidone solid dispersion tablets, dissolution increased linearly and reached 93.4% after 2 hr. Reducing the volume of test medium from 900 to 300 ml markedly decreased the dissolution rate of the tablets containing 1 : 20 HPCD solid dispersions and 1 : 10 copolyvidone solid dispersion. For 1 : 20 copolyvidone solid dispersion tablets, there was no significant change in dissolution rate up to 1 hr with different volumes of test medium. Preparation of the copolyvidone solid dispersion (1 : 20) in capsules markedly delayed the dissolution (31.2 % after 2hr) due to the limited diffusion within capsules. The permeation rate $(13.4\;g/cm^2\;after\;8\;hr)$ of DDB through rabbit duodenal mucosa from copolyvidone solid dispersion (1 : 10) was markedly enhanced, when compared with drug alone or physical mixtures. From overall findings, DDB formulations containing copolyvidone solid dispersions (1 : 20) could be used to remarkably improve the dissolution rate in dosage form of powders and tablets.
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