• Journal of Nutrition and Health
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• v.28 no.5
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• pp.415-425
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• 1995

This study was performed to investigate the effect of dietary protein level on the metabolic changes of Ca and skeletons in postmenopausal women, using ovariecotomized rats as an animal model. The female rats of 200∼250g were fed either 8%(L) or 50%(H) casein diet for 15 weeks(1st experiment). At 15th week, the rats of each diet group were undergone ovariectomy or sham-operation and they were continued to feed the same experimantal diet for 9 more months(2nd experiment). Ca metabolism, kidney function and bone composition were determined at the end of 1st experiment, 3rd and 9th month of 2nd experiment. After 1st experiment, high protein group showed higher urinary Ca and protein excretion, however, there was no difference in GFR and urinary hydroxyproline excretion. The weights, ash and Ca content of femur, scapular and vertebra tended to be higher in high protein groups which tells that high protein promoted skeletal growth. In 2nd experiment, high protein group showed higher urinary Ca and protein excretion and lower Ca absorption and balance. GFR was not affected by dietary protein and ovariectomy but increased with time, as well as kidney weight which shows the continuous development of kidney at this age of 15 month in rats. There were no difference in urinary hydroxyproline, serum ALP, and PTH among experimental groups. The weights of femur, scapular, 4th vertebra increased with time, showing the skeleton continues to grow at this age in rats. However, Ca contents, Ca/wt, Ca/ash were decreased with time and tended to be lower in high protein group especially in femur. In conclusion, prolonged feeding of high protein diet deteriorated Ca metabolism and induced bone loss as time after menopause is extended.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.3
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• pp.414-421
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• 2014

This study was conducted to determine the effects of saturated long-chain fatty acids (LCFA) on cell proliferation and triacylglycerol (TAG) content, as well as mRNA expression of ${\alpha}s1$-casein (CSN1S1) and genes associated with lipid and protein synthesis in bovine mammary epithelial cells (BMECs). Primary cells were isolated from the mammary glands of Holstein dairy cows, and were passaged twice. Then cells were cultured with different levels of palmitate or stearate (0, 200, 300, 400, 500, and 600 ${\mu}M$) for 48 h and fetal bovine serum in the culture solution was replaced with fatty acid-free BSA (1 g/L). The results showed that cell proliferation tended to be increased quadratically with increasing addition of stearate. Treatments with palmitate or stearate induced an increase in TAG contents at 0 to 600 ${\mu}M$ in a concentration-dependent manner, and the addition of 600 ${\mu}M$ was less effective in improving TAG accumulation. The expression of acetyl-coenzyme A carboxylase alpha, fatty acid synthase and fatty acid-binding protein 3 was inhibited when palmitate or stearate were added in culture medium, whereas cluster of differentiation 36 and CSN1S1 mRNA abundance was increased in a concentration-dependent manner. The mRNA expressions of peroxisome proliferator-activated receptor gamma, mammalian target of rapamycin and signal transducer and activator of transcription 5 with palmitate or stearate had no significant differences relative to the control. These results implied that certain concentrations of saturated LCFA could stimulate cell proliferation and the accumulation of TAG, whereas a reduction may occur with the addition of an overdose of saturated LCFA. Saturated LCFA could up-regulate CSN1S1 mRNA abundance, but further studies are necessary to elucidate the mechanism for regulating milk fat and protein synthesis.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.837-845
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• 2014

Background: Overweight and obesity are recognized as major drivers of cancers including breast cancer. Several cytokines, including interleukin-6 (IL-6), IL-10 and lipocalin 2 (LCN2), as well as dysregulated cell cycle proteins are implicated in breast carcinogenesis. The nuclear, casein kinase and cyclin-dependent kinase substrate-1 (NUCKS-1), is a nuclear DNA-binding protein that has been implicated in several human cancers, including breast cancer. Objectives: The present study was conducted to evaluate NUCKS-1 mRNA expression in breast tissue from obese patients with and without breast cancer and lean controls. NUCKS-1 expression was correlated to cytokine profiles as prognostic and monitoring tools for breast cancer, providing a molecular basis for a causal link between obesity and risk. Materials and Methods: This study included 39 females with breast cancer (G III) that was furtherly subdivided into two subgroups according to cancer grading (G IIIa and G IIIb) and 10 control obese females (G II) in addition to 10 age-matched healthy lean controls (G I). NUCKS-1 expression was studied in breast tissue biopsies by means of real-time PCR (RT-PCR). Serum cytokine profiles were determined by immunoassay. Lipid profiles and glycemic status as well as anthropometric measures were also recorded for all participants. Results: IL-6, IL-12 and LCN2 were significantly higher in control obese and breast cancer group than their relevant lean controls (p<0.05), while NUCKS-1 mRNA expression was significantly higher in the breast cancer group compared to the other groups (p<0.05). Significant higher levels of IL-6, IL-12, and LCN2 as well as NUCKS-1 mRNA levels were reported in G IIIb than G IIIa, and positively correlated with obesity markers in all obese patients. Conclusions: Evaluation of cytokine levels as well as related gene expression may provide a new tool for understanding interactions for three axes of carcinogenesis, innate immunity, inflammation and cell cycling, and hope for new strategies of management.

• Preventive Nutrition and Food Science
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• v.7 no.2
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• pp.139-145
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• 2002

Hen egg-white lysozyme, ovalbumin, egg-yolk phosvitin, acid-precipitated soy protein and $\alpha$$_{sl}$ milk casein were covalently linked with galactomannan through a controlled dry-heating at 6$0^{\circ}C$ under 79% relative humidity without any chemical reagent. Neoglycosylation by the covalent binding of polysaccharide chains brought a significant improvement into the surface functionalities of food proteins. Excellent emulsifying properties and foaming properties were observed in all protein-galactomannan conjugates. Bacterial mutagenesis tests and animal dose test were done to evaluate the food safety of the protein-galactomannan conjugates. The neo-glycoproteins were negative for Ames test using Salmonella typhimurium TA100 (hisG46) and TA98 (hisD3052) strains, and rec-assay using Bacillus subtilis Hl7 (rec) and M45 (re $c^{+}$) strains. All substances were also nontoxic for oral administration to rats. L $D_{50}$ 's of these substances were all more than 7.5 g/kg body-weight of rat. No effect was also observed in the weight increases and the concentrations of total cholesterol, triglyceride and phospholipids in blood serum of the administrated rats with 7.5 g/kg conjugates. Thus, Maillard-type protein-polysaccharide conjugates prepared by covalent attachment of galactomannan to food proteins were proposed to be useful as a safe functional biopolymer in this study.y.

• Journal of Dairy Science and Biotechnology
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• v.30 no.1
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• pp.17-22
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• 2012

Since the prevalence of allergies is increasing, food allergy is a major concern for consumers, as well as for the food industry. The foods that account for over 90% of all moderate to severe allergic reactions to food are milk, eggs, peanuts, soybeans, fish, shellfish, wheat, and tree nuts. Of these food allergens, milk is one of the major animal food allergens in infants and young children. Milk is the first food that an infant is exposed to; therefore, the sensitization rate of milk in sensitive individuals is understandably higher. The mechanisms involved in allergic reactions caused by this hypersensitivity are similar to those of other immune-mediated allergic reactions. The reactions occur in the gastrointestinal tract, skin, and respiratory tract, with headaches and psychological disorders occurring in some instances. The major allergenic proteins in milk are casein, ${\beta}$-lactoglobulin, and ${\alpha}$-lactalbumin, while some of the minor allergenic proteins are lactoferrin, bovine serum albumin, and immunoglobulin. Reliable allergen detection and quantification are essential for compliance with food allergen-labeling regulations, which protect the consumer and facilitate international trade.

• Journal of Microbiology and Biotechnology
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• v.6 no.1
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• pp.1-6
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• 1996

A bacterial strain NS70 producing an alkaline protease was isolated from soil samples taken near a hot spring and identified as Bacillus licheniformis by its morphological and physiological properties and cellular fatty acid analysis. The isolated alkaline protease was purified by ammonium sulfate fractionation, DEAE-, CM-, and Phenyl-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 32, 000 Da by sodium dodecylsulfate polyacrylamide gel electrophoresis. Its optimal pH and temperature for proteolytic activity against Hammarsten casein were 12 and $65^{\circ}C$, respectively. The enzyme was stable at alkaline pH range from 6.0 to 12.0, and fairly stable up to $65^{\circ}C$. The enzyme was inhibited by phenylmethylsulfonyl fluoride but not by EDTA and N-ethylmaleimide indicating that the enzyme is serine protease. Enzyme activity was markedly inhibited by $Hg^{2+}$ and $Cu^{2+}$. Autolytic phenomena were observed on purified protease NS70 but autolysis was reduced by the addtion of $Ca^{2+}$ ion or bovine serum albumin.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.852-858
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• 2008

An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.

• Korean Journal of Agricultural Science
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• v.45 no.4
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• pp.635-643
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• 2018

Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.74-74
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• 2002

Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

• Journal of Nutrition and Health
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• v.4 no.4
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• pp.15-23
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• 1971

On the assumption that the supplementation of certain nutrients or foods to the rice diet (low protein, low fat, and low vitamins) may decrease, to some extent, the degree of suffering from abnormal environments, such as vibration, noises, gases, dusts, smog etc. a series of experiments were started. As the first report the nutrition under vibration was studied in this experiment. Sixty (60) young growing male rats weighing about 65 grams were used, grouping to five (5) groups, twelve (12) rats each group. They were fed on the following five (5) experimental diets: rice diet (basal diet), rice diet+casein, rice diet+vitamins, rice $diet+{\alpha}-tocopherol$, and rice diet+ginseng powder (see the tables 1 and 2) for the period of 14 weeks experiment. During the experiment period the half number of the rats of each group were exposed to the three (3) hours vibration every day. The protective effect of each diet against the vibration may be summarized as follows. 1. The growth of rice diet group was impaired significantly under vibration, However, those of other groups (protein-supplemented, vitamin-supplemented, ${\alpha}-tocopherol-supplemented$ and ginseng-supplemented groups) were impaired much less compared with rice diet group. 2. The feed efficiency of the rice diet group was decreased significantly under vibration. It is estimated that the biological availability of nutrients was impaired under this environment. On the other hand, the feed efficiencies of protein supplemented, of vitamin supplemented, and of ginseng supplemented groups were not decreased under vibration, statistically. 3. There is tendency that the food spillages of vibration groups are higher than those of non-vibration groups. Especially it seems true in the case of rice diet group. The food spillage may be, to some extent, related with mental nervousness of animals. From the point that the food spillage of ginseng supplemented group is significantly lower than those of other groups it is thought ginseng acts some good role in protecting nervous system from suffering from vibration. 4. In all groups except protein supplemented group, liver fat of vibration group tends to be higher than that of non-vibration group. 5. It shows that, in general, the serum alkaline phosphatase activity of the vibration group is significantly higher than that of the non-vibration group. It seems that there may be, to some extent, corelation between the amount of liver fat and serum alkaline phosphatase activity. 6. There is tendency that, in rice diet group, the organs of vibration group are smaller than those of non-vibration group, especially lung is so. It is thought that this may be due to the poor growth of whole body size in vibration group.