• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.218-224
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• 2018

Objective: This study evaluated the effects of different levels of protein concentrate supplementation on the growth performance of yak calves, and correlated the growth rate to changes occurring in the plasma- amino acids, -insulin profile, and signaling activity of mammalian target of rapamycin (mTOR) cascade to characterize the mechanism through which the protein synthesis can be improved in early weaned yaks. Methods: For this study, 48 early (3 months old) weaned yak calves were selected, and assigned into four dietary treatments according to randomized complete block design. The four blocks were balanced for body weight and sex. The yaks were either grazed on natural pasture (control diet) in a single herd or the grazing yaks was supplemented with one of the three protein rich supplements containing low (17%; LP), medium (19%; MP), or high (21%; HP) levels of crude proteins for a period of 30 days. Results: Results showed that the average daily gain of calves increased (0.14 vs 0.23-0.26 kg; p<0.05) with protein concentrates supplementation. The concentration of plasma methionine increased (p<0.05; 8.6 vs $10.1-12.4{\mu}mol/L$), while those of serine and tyrosine did not change (p>0.05) when the grazing calves were supplemented with protein concentrates. Compared to control diet, the insulin level of calves increased (p<0.05; 1.86 vs $2.16-2.54{\mu}IU/mL$) with supplementation of protein concentrates. Addition of protein concentrates up-regulated (p<0.05) expression of mTOR-raptor, mammalian vacuolar protein sorting 34 homolog, the translational regulators eukaryotic translation initiation factor 4E binding protein 1, and S6 kinase 1 genes in both Longissimus dorsi and semitendinosus. In contrast, the expression of sequestosome 1 was down-regulated in the concentrate supplemented calves. Conclusion: Our results show that protein supplementation improves the growth performance of early weaned yak calves, and that plasma methionine and insulin concentrations were the key mediator for gene expression and protein deposition in the muscles.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.

• Reproductive and Developmental Biology
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• v.34 no.4
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• pp.275-281
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• 2010

The aim of this study was to investigate what protein(s) of porcine epididymal fluid (pEF) are able to enhance the nuclear maturation of porcine germinal vesicle (GV) oocytes in vitro. Proteins of pEF were fractionated by affinity, ion exchange, and gel filtration chromatography. Porcine cumulus-oocytes complexes (COC) from follicles were cultured in tissue culture medium (TCM 199) containing various fractions obtained by chromatography. Porcine COCs were also cultured in TCM 199 containing various meiosis inhibitors and pEF. After 24 or 48 h culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. When porcine COCs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than 80% of oocytes were unable to resume meiosis. However, porcine COCs supplemented with pEF were able to overcome the inhibitory effect of dbcAMP and Fo. Maturation rate of oocytes was significantly (p<0.05) increased in the media supplemented with cationic protein(s) during in vitro maturation than in those with anionic protein(s) (44.1% vs 20.0%). When oocytes were cultured in the TCM 199 with fractions obtained by gel filtration, the maturation rate of oocytes was significantly (p<0.05) higher in fraction 11 containing 18 kDa than other fractions. The present study suggests that 1) dbcAMP and Fo prevent the spontaneous maturation of oocyte after isolation from follicles, and that pEF contain a substance(s) that improves meiosis resumption in vitro of porcine COCs, 2) cationic 18 kDa protein(s) are responsible for promotion of Mil stage.

• KSBB Journal
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• v.4 no.2
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• pp.65-68
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• 1989

In order to determin the optimum extraction condition for protein in a Chrysalis of Silk Worm, Temperature, Time, and pH variation were conducted in the extraction. N.S.I. and nitrogen contents of the extracts in this condition were identified by Kjeldaha method and a C.H.N. corder. The results were as follows; Crude protein and crude fat contents in the extracts were 23.34% and 15.61%, respectively. N.S.I. values with respect to the temperature variation were 6.7% at 6$0^{\circ}C$, 7.94% at 8$0^{\circ}C$ and 8.67% at 10$0^{\circ}C$ in the condition of pH 4, 1 hr. N.S.I. values in pH variation were 7.94% at pH4, 8.99% at pH6, 9.70% at pH8, 10.19% at pH 10 and 12.16% at pH 12 in the condition of 8$0^{\circ}C$, 1hr. N.S.I. values in extraction time variation were 8.67% in 1hr, 9.23% in 2hr and 9.76% in 3hr in the condition of 10$0^{\circ}C$, pH 4. The tendency of N.S.I. variation in this extration condition was reconfirmed also by a C.H.N. corder.

• BMB Reports
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• v.34 no.6
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• pp.559-565
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• 2001

Protein carboxyl O-methyltransferase (E.C.2.1.1.24) may play a role in the repair of aged protein that is spontaneously incorporated with isoaspartyl residues. The porcine brain carboxyl O-methyltransferase was cloned in the pET32 vector, and overexpressed in E.coh (BL21) that harbors pETPCMT, which encodes 227 amino acids, including tagging proteins at the N-terminus. The protein sequence of the cloned porcine brain PCMT (r-pbPCMT) shares a 98% identity with that of human erythrocyte PCMT and rat brain PCMT. It is 100% identical with that of bovine brain. The r-pbPCMT was purified using Ni-NTA affinity chromatography and digested by enterokinase in order to remove the protein tags. Then Superdex 75HR gel filtration chromatography was performed. The r-pbPCMT exhibited similar in vitro substrate specificities with the PCMT that was purified from porcine brain. The molecular weight of the enzyme was estimated to be 24.5 kDa on the SDS polyacrylamide gel electrophoresis. The $K_m$ value was $1.1{\times}10^{-7}\;M$ for S-adenosyl-L-methionine. S-adnosyl-L-homocysteine was a competitive type of inhibitor with the $K_i$ value of $1.38{\times}10^{-4}\;M$. The enzyme has optimal activity at pH 6.0 and $37^{\circ}C$. These results indicate that the expressed enzyme is functionally similar to the natural protein. It also suggests that it may be a suitable model to further understand the function of the mammalian enzyme.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.3
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• pp.335-340
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• 2012

The data used came from two trials undertaken under the same climatic conditions (spring-summer). In both trials pluriparious buffaloes were utilized similar in weight, body condition score, and milk production from the previous year. From the first trial the data used was from the sub-period 23-88 DIM provided by seven animals fed ad libitum with diet A (6.69 MJ/kg DM; 158.30 g/kg of crude protein) with a forage/concentrate ratio of 48/52. From the second trial the data used was from the sub-period 33-90 DIM provided by seven animals fed ad libitum with diet B (6.63 MJ/kg DM; 179.50 g/kg of crude protein) and by seven animals fed ad libitum with diet C (5.99 MJ/kg DM; 155.40 g/kg of crude protein), each of the diets had the same forage/concentrate ratio (53/47). A significant difference was found in milk production between group B and C (13.08 vs. 11.56 kg/d, p<0.05), an intermediate production (12.10 kg/d) was noted in group A. A significant difference was found between fat (76.58 vs. 69.24 g/kg, p<0.05), protein (46.14 vs. 43.16 g/kg, p<0.05) and casein (39.94 vs. 34.98 g/kg, p<0.05) of the milk of group B with respect to group A. The milk of group C gave fat values (71.80 g/kg), protein (45.52 g/kg) and casein (39.06 g/kg) statistically equal to those of group B. The milk of groups B and C, in respect to the milk of group A, gave values of $K_{20}$ (1.77, 1.82 vs. 3.68 min, p<0.05), statistically lower and values of $A_{30}$ (48.28, 47.27 vs. 40.64 mm, p<0.05) statistically higher. Two simple linear regressions were calculated where the independent variable (x) was the daily standardized milk production, the dependent variable (y) or the daily intake of net energy or crude protein. Equation 1) NE (MJ/d) = 74.4049+2.8308${\times}$kg of normalized milk; equation 2) CP (kg/d) = 1.4507+0.1085${\times}$kg of normalized milk, both the equations were significant (p<0.05) with determination coefficients of 0.58 and 0.50 respectively. For a production of normalized milk that varies from 9 to 13 kg, the respective energy-protein concentrations fluctuate from 6.09 to 6.78 MJ/kg DM and from 148.00 to 174.46 g/kg DM.

• The Microorganisms and Industry
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• v.19 no.2
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• pp.2-10
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• 1993

이 총론에서는 cell cycle의 조절에 관계하는 p34cdc kinase의 특성과 기질 그리고 cell cycle에서의 역할을 살펴보고, 또 cell cycle에서와 여러가지 세포내의 현상에 중요한 역할을 하는 것으로 알려진 casein kinase II의 특성과 기질 그리고 cell cycle에서의 역할을 살펴보고자 한다. 그리고 이런 효소들을 연구하는 데 필수적인 방법이나 시약들로 소개하고자 한다.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.1
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• pp.34-37
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• 2002

Two experiments were conducted to evaluate the nutritional value of common fodder tree leaves with goats fed at 50% of total ration. In experiment 1, leaves from six fodder tree species i.e., Ailanthus aitissima, Elaeagnus angustifolic, Morus alba (Mulberry), Populus spp, Robina pseudoacacia and Salix babylonia were harvested in spring and winter from northern areas of Pakistan. Chemical composition and apparent in situ dry matter digestibility (DMD) of fodder tree leaves were measured. Results showed that crude protein (CP) values were higher (p<0.05) in all the species during spring compared to winter (17.9% vs 12.0%). The concentration of NDF in Elaeagnus and Robinia was higher in spring, whereas no seasonal difference was found in other species. In situ DMD was higher (p<0.05) in Ailanthus and Populus at spring while it was higher (p<0.05) in Elaeagnus, Mulberry and Robinia at winter. There was no (p<0.05) seasonal effect on in situ DMD of Salix. In experiment II, four iso-nitrogenous and iso-caloric rations viz., A, B, C and D were prepared containing 50% (winter harvested) sun dried leaves of Salix, Robinia, Mulberry and Elaeagnus, respectively and 50% concentrate. Dry matter and crude protein intakes were higher (p<0.05) given ration A (Salix) whereas DM and CP digestibility was lowest (p<0.05) given ration B. Nitrogen retention was higher (p<0.05) given ration A. Goats fed on fodder tree leaves and concentrate showed moderate intake and digestibility.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.251-259
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• 1990

Asp. usumii IAM 2185 was selected as a strain producing the powerful raw starch digesting glucoamylase. The optimum initial pH, the optimum temperature and the optimum cultural time for the enzyme production on wheat bran medium were pH 6-8,25-$30^{\circ}C$ and 72 hrs, respectively. The addition of ammonium nitrate and albumin on wheat bran medium, respectively, increase slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation, CM-cellulose and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 34.3 U/mg protein and the yield of enzyme activity was 10.3%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 67,000 by SDS polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pR 3.7. The optimum temperature and optimum pH were $60^{\circ}C$and pH 3.0 and the purified enzyme was stable in the pH range of 1.0-11.0. The purified enzyme was stable below $50^{\circ}C$ and its thermostability was greatly increased by the addition of $Ca^{2+}$. The purified enzyme showed a high hydrolysis rate on various raw starches such as corn, rice, yam, arrow root, sweet potato and glutinous rice.

• Mycobiology
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• v.34 no.3
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• pp.128-130
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• 2006

The solid waste of sago industry using cassava was fermented by Aspergillus niger, Aspergillus terreus and Rhizopus stolonifer in solid state fermentation. Cassava waste contained 52 per cent starch and 2.9 per cent protein by dry weight. The amylase activity was maintained at a high level and the highest amylase activity was observed on the $8^{th}$ day in R. stolonifer mediated fermentation. R. stolonifer was more efficient than Aspergillus niger and Aspergillus terreus in bioconverting cassava waste into fungal protein (90.24 mg/g) by saccharifying 70% starch and releasing 44.5% reducing sugars in eight days of solid state fermentation.