• Title/Summary/Keyword: Small protein

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Licochalcone C Induces Autophagy in Gefitinib-sensitive or-resistant Human Non-small Cell Lung Cancer Cells (Gefitinib-민감성 또는 내성 비소세포폐암 세포에서 Licochalcone C에 의한 자가포식 유도)

  • Oh, Ha-Na;Yoon, Goo;Chae, Jung-Il;Shim, Jung-Hyun
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1305-1313
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    • 2019
  • Licochalcone (LC), isolated from the roots of Glycyrrhiza inflata has multiple pharmacological effects including anti-inflammatory and anti-tumor activities. To date, Licochalcone C (LCC) has induced apoptosis and inhibited cell proliferation in oral and bladder cancer cells, but lung cancer has not yet been studied. In addition, no study reported LCC-induced autophagy in cancer until now. The present study was designed to investigate the effect of LCC on gefitinib-sensitive and -resistant lung cancer cells and elucidate the mechanism of its action. The 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay data showed that LCC significantly inhibited cell viability in non-small cell lung cancer (NSCLC) HCC827 (gefitinib-sensitive) and HCC827GR (gefitinib-resistant) cell lines. Interestingly, Annexin V/7-aminoactinomycin D double staining and cell cycle analysis showed an apoptosis rate within about 20% at the highest concentration of LCC. LCC induced G2/M arrest by reducing the expression of the cell cycle G2/M related proteins cyclin B1 and cdc2 in NSCLC cell lines. Treatment of LCC also induced autophagy by increasing the expression of the autophagy marker protein microtubule-associated protein 1 light chain 3 (LC3) and the protein autophagy-related gene 5 involved in the autophagy process. In addition, LCC increased the production of reactive oxygen species (ROS), and the cell viability was partially restored by treatment with the ROS inhibitor N-acetyl-L-cysteine. In western blotting analysis, the expression of cdc2 was increased and LC3 was decreased by the simultaneous treatment of NAC and LCC. These results indicate that LCC may contribute to anti-tumor effects by inducing ROS-dependent G2/M arrest and autophagy in NSCLC. In conclusion, LCC treatment may be useful as a potential therapeutic agent against NSCLC.

Production Increase of Milk in Dairy Cow by Metabolic Profile Test (대사판정시험을 이용한 젖소의 우유증산)

  • Lee Chang-Woo;Kim Bonn-Won;Ra Jeong-Chan;Shin Sang-Tae;Kim Doo;Kim Jong-Taik;Hong Soon-Il
    • Journal of Veterinary Clinics
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    • v.10 no.1
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    • pp.65-94
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    • 1993
  • This study examined metabolic profiles of 1349 Holstein cows from 91 commercial herds. Thirteen parameters which are consisted of twelve blood components and body condition score were examined and their mean values. standard deviations and standard limits, which are 80% confidential limits, in each lactational stage were reported. The variations of each parameter affected by season, individual milk yield, adjusted corrected milk yield of herd. and lactation number were also reported. A model of metabolic profile test applicable to this country where the average number of cows in a herd is small as to be fifteen is designed. Metabolic profiles as reflected in each parameter were discussed in relation to adequacy of dietary intake for production, milk production, reproductive performance, and diseases, and the possible measure to improve productivity of dairy cows were proposed. Much of the variation in parameters was due to differences between herds, and less to differences between seasons, differences between individual milk yield, and differences between lactational stages. As the average herd size in this country is small, it is believed that all the cows in a herd must be sampled, and the individual result of each parameter was compared with the standard limit for each lactational stage, and the percentage of cows which are outside the standard limits in a herd was calculated to use as a criteria for evaluation of the herd. Data outside the 99% confidential limits were to be deleted at first, but when the trends of the data outside the 99% confidential limits are same as the trends of the data within 99% confidential limits, the deleted data must be reviewed again, otherwise some important informations would be missed. The mean concentration of blood urea nitrogen in this study was much higher than that was reported in England, U.S.A. and Japan, and it was similar to the upper limits reported in England, U.S.A. and Japan. So it was thought that the concentration of blood urea nitrogen is improper as a criteria for protein intake. The increase of serum total protein cocentration beyond standard limits was due to increase of serum globulin concentration in most of the cows. The correlation coefficient between serum and protein and serum globulin concentration was 0.83. Serum globulin concentration was negatively related to adjusted corrected milk of herd. Serum albumin, calcium and magnessium concentrations were negatively related to adjusted corrected milk of herd, which indicate that high-producing individual or high-producing herd have not taken sufficient protein/amino acids, calcium and magnessium. Packed cell volume was negatively related to adjusted corrected milk of the herd, and the trend was same In each lactational stage. The correlation coefficient between serum and packed cell volume was 0.16 and the correlation was very weak. Blood glucose concentration was lowest in early lactational stage, which indicates negative energy balance in early lactational stage. Blood glucose concentration was negatively related to adjusted corrected milk of herd from peak to late lactational stage, which indicates negative energy balance during the period in high-producing individuals or high-producing herds. Correlation coefficient between serum aspartate aminotransferase activity and serum ${\gamma}$-glutamyltransferase activity was 0.41, and this indicates that serum ${\gamma}$-glutamyltransferase should be included as a parameter of metabolic profile test to evaluate liver function. Body condition score of dairy cows in this country was lower than that of Japan in every lactational stages, and the magnitude of increase in body condition score during middle and late lactational stages was small. Metabolic profile can not be evaluated with solely nutritional intake. When an individual or large percentage of cows in a herd have adnormal values In parameters of metabolic profile test, veterinary clinician and nutritionist should cooperate so as to diagnose diseases and to calculate the e of no운ents simultaneously.

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Varietal Response to Grain Quality and Palatability of Cooked Rice Influenced by Different Nitrogen Applications (질소 시비조건에 따른 벼 품종의 미질과 식미특성 반응)

  • Kim, Jeong-Il;Choi, Hae-Chun;Kim, Kwang-Ho;Ahn, Jong-Kuk;Park, No-Bong;Park, Dong-Soo;Kim, Chun-Song;Lee, Ji-Yoon;Kim, Jae-Kyu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.1
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    • pp.13-23
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    • 2009
  • To intensively analyze and examine the changes in palatability of cooked rice and structural properties of rice grains influenced by increased nitrogen topdressing during the panicle formation and ripening stage, a series of experiments were carried out using three japonica rice cultivars with contrasting rice quality characteristics. The protein content of brown rice increased maximally up to $35{\sim}47%$ of that in standard N6 plot with the increase in nitrogen topdressing fertilizer during 20 days before heading to 10 days after flowering. The high-yielding rice cultivars showing poor palatability of cooked rice revealed larger increase in protein content of rice grains by increased nitrogen topdressing. Under the same nitrogen level of 15 kg per 10a with nitrogen topdressing at 3 kg/10a, high-yielding rice cultivars, Yumehikari and Reihou showed the significant increase in protein content of brown rice when topdressing was applied at 10 days after flowering as compared with when it was applied at 30 days after transplanting. Although the variation in amylose content of milled rice as affected by nitrogen topdressing level was relatively small, it decreased within 1% with the opposite tendency against increased protein content of brown rice by increased nitrogen topdressing. The total score of sensory evaluation was higher in the order of Hinohikari < Yumehikari < Reihou in panel test. It decreased significantly by increased amounts of nitrogen topdressing during 20 days before heading to 10 days after flowering when nitrogen level was higher than 12 kg/10a. The more poor palatable rice cultivar in panel test revealed the larger decreasing in total score of sensory evaluation by higher nitrogen topdressing rates. All sensory evaluation components were largely affected by the change in protein content of brown rice rather than amylose content of milled rice. The influence of protein content to palatability of cooked rice was larger in poor-palatable rice than in high-palatable rice. The protein content decreased drastically from outer layer to inner layer of rice grains, while the amylose content increased on the contrary. The high-palatable rice exhibited higher distribution of protein content on bran layer but lower distribution of protein content on the layer of polished rice as compared with the poor-palatable rice. Especially, the high-palatable rice showed also significantly lower distribution of amylose content on the outer layer of polished rice as compared with the poor-palatable rice.

Production of the Polyclonal Antibody That Recognizes the Mutant M Protein of Japanese Encephalitis Virus: Role of Its Charged Residues in Virus Production (일본뇌염바이러스의 Mutant M 단백질에 반응하는 다클론항체의 생산: 극성 아미노산 잔기의 바이러스 생산과정에서의 역할)

  • Kim, Jeong-Min;Yun, Sang-Im;Song, Byung-Hak;Kim, Jin-Kyoung;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.140-147
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    • 2010
  • Japanese encephalitis virus (JEV), a member of the mosquito-borne flaviviruses, causes epidemics of viral encephalitis in the Southeastern Asia. JEV is a small enveloped virus with a positive-sense RNA genome; the infectious virion consists of three structural proteins, namely capsid, membrane (M; a mature form of its prM precursor), and envelope proteins. Here, we investigated a role of the charged residues found at the N-terminus of the JEV M protein in virus production. Using an infectious JEV cDNA, we generated two mutant cDNAs, Mm1 and Mm2, by charged-to-alanine substitution for $E^9$ and $K^{15}K^{16}E^{17}$ residues of the M protein, respectively. By transfection of wild-type or each of the two mutant RNAs transcribed from the corresponding cDNAs, we found that Mm2, but not Mm1, had a ~3-log decrease in virus production, even though a comparable amount of all three structural proteins were produced in transfected cells. Interestingly, the prM protein expressed in Mm2 RNA-transfected cells was not recognized by the polyclonal antiserum raised against the N-terminal 44 amino acids of the wild type M protein, but reacted to the antiserum raised against the corresponding region of the mutant Mm2. Our results indicate that three charged residues ($K^{15}K^{16}E^{17}$) in JEV M protein play a role in virus production. Two polyclonal antisera specifically recognizing the wild-type or Mm2 version of the M protein would provide a useful reagent for the functional study of this protein in the virus life cycle.

Expression of ERCC1, MSH2 and PARP1 in Non-small Cell Lung Cancer and Prognostic Value in Patients Treated with Platinum-based Chemotherapy

  • Xie, Ke-Jie;He, Hong-Er;Sun, Ai-Jing;Liu, Xi-Bo;Sun, Li-Ping;Dong, Xue-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2591-2596
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    • 2014
  • Purpose: To evaluate the prognostic value of the expression of excision repair cross-complementation group l (ERCC1), MutS protein homolog 2 (MSH2) and poly ADP-ribose polymerase 1 (PARP1) in non-small-cell lung cancer patients receiving platinum-based postoperative adjuvant chemotherapy. Methods: Immunohistochemistry was applied to detect the expression of ERCC1, MSH2 and PARP1 in 111 cases of non-small cell lung cancer paraffin embedded surgical specimens. Through og-rank survival analysis, we evaluated the prognostic value of the ERCC1, MSH2, PARP1 and the related clinicopathological factors. COX regression analysis was used to determine whether ERCC1, MSH2 and PARP1 were independent prognostic factors. Results: In the enrolled 111 non-small cell lung cancer patients, the positive expression rate of ERCC1, MSH2 and RARP1 was 33.3%, 36.9% and 55.9%, respectively. ERCC1 (P<0.001) and PARP1 (P=0.033) were found to be correlated with the survival time while there was no correlation for MSH2 (P=0.298). Patients with both ERCC1 and PARP1 negative cancer had significantly longer survival time than those with ERCC1 (P=0.042) or PARP1 (P=0.027) positive alone. Similalry, the survival time of patients with both ERCC1 and PARP1 positive cancer was shorter than those with ERCC1 (P=0.048) or PARP1 (P=0.01) positive alone. Conclusion: Patients with ERCC1 or PARP1 negative non-small cell lung cancer appear to benefit from platinum-based postoperative adjuvant chemotherapy.

The Effects of Chinese and Argentine Soybeans on Nutrient Digestibility and Organ Morphology in Landrace and Chinese Min Pigs

  • Qin, G.X.;Xu, L.M.;Jiang, H.L.;van der Poel, A.F.B.;Bosch, M.W.;Verstegen, M.W.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.555-564
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    • 2002
  • Twenty Landrace and twenty Min piglets, with an average initial body weight of 22.4 kg, were randomly divided into 5 groups with 4 animals per group, within each of the breeds. The piglets were housed in individual concrete pens. Each group of the piglets was fed one of 5 diets. The diets contained either 20% raw Argentine soybeans, 20% processed Argentine soybeans ($118^{\circ}C$ for 7.5 min.), 20% raw Chinese soybeans, 20% processed Chinese soybeans ($118^{\circ}C$ for 7.5 min.) or no soybean products (control diet). Faecal samples were collected on days 6, 7 and 8 of the treatment period. Digestibilities of dietary nutrients were determined with AIA (acid insoluble ash) as a marker. After a 17 day treatment, three piglets were killed from each of the groups. Tissue samples of small and large intestine for light and electron microscopy examination were taken immediately after the opening of abdomen. Then, the weight or size of relevant organs was measured. The results show that the digestibilities of dry matter (DM), crude protein (CP) and fat were higher in Min piglets than in Landrace piglets (p<0.05). The diets containing processed soybeans had a significant higher CP digestibility than the control diet and the diets containing raw soybeans (p<0.05). Landrace piglets had heavier and longer small intestines, heavier kidneys and a lighter spleen than Min piglets (p<0.05). The pancreas of the animals fed the diets containing processed soybeans was heavier than that of the animals fed control diet (p<0.05) and the diets containing raw soybeans. But, the differences between raw and processed soybean diets were not significant. A significant interaction (p<0.05) between diet and pig breed was observed in weight of the small intestine. The Landrace piglets increased the weight in their small intestine when they were fed the diets containing soybeans. In the light micrographs and electron scanning micrographs, it was found that the villi of small intestinal epithelium of animals (especially Landrace piglets) fed the diets containing raw Chinese soybeans were seriously damaged. The transmission electron micrograph showed that a lot of vesicles were located between the small intestinal microvilli of these piglets. The histological examination also indicated that the proportion of goblet cells in villi and crypts in the piglets consuming the control diet was significantly lower (p<0.01 and p<0.02, respectively) than those of the animals consuming the diets containing raw or processed soybeans.

Inhibition of Starch Biosynthesis by Antisense Expression of cDNAs Encoding ADP-Glucose Pyrophosphorylase Small Subunit in Sweetpotato (고구마에서 ADP-Glucose Pyrophosphorylase Small Subunit cDNA의 Antisense 발현에 의한 전분생합성 저해)

  • Min, Sung-Ran;Bae, Jung-Myung;Harn, Chee-Hark;Jeong, Won-Joong;Lee, Young-Bok;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.277-283
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    • 2007
  • Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.

Quality and Physicochemical Characteristics of Small Black Soybean Cultivar Cultivated in the North-central Region (중북부지역에서 재배한 소립 검정콩의 품질 및 이화학 특성)

  • Kim, Hyun-Joo;Jung, Gun Ho;Lee, Ji Hae;Lee, Byong Won;Lee, Yu Young;Kim, Sung Kook;Lee, Byoung Kyu;Woo, Koan Sik
    • The Korean Journal of Food And Nutrition
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    • v.31 no.6
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    • pp.792-801
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    • 2018
  • Proximate compositions, quality and physicochemical characteristics of small black soybean cultivar, cultivated in the north-central region of South Korea with different seeding periods, were evaluated. Proximate compositions, chromaticity, water binding capacity, water solubility index, swelling power, and antioxidant properties were significantly different among cultivars and different seeding periods. Moisture, crude ash, fat, protein, and carbohydrate contents of small black soybean cultivar were 5.53~6.69, 5.47~6.54, 15.38~19.14, 34.17~40.26, and 32.04~36.85 g/100 g, respectively. Lightness, redness and yellowness were 35.60~38.61, -0.02~0.07 and -0.56~-0.13, and water binding capacity, water solubility index and swelling power were 84.48~148.31, 46.65~54.89 and 29.87~35.12%, respectively. Total polyphenol contents of first, second, and third seedings on small black soybean cultivar were 10.40~15.48, 9.86~14.85 and 8.61~15.39 mg GAE/g; total flavonoid contents were 5.81~7.25, 5.81~7.34 and 5.52~7.64 mg CE/g, respectively. DPPH radical scavenging activity was 4.55~7.86, 3.99~8.79, and 3.74~9.43 mg TE/g, and ABTS radical scavenging activity was 9.32~12.90, 8.64~13.39, and 8.51~14.35 mg TE/g, respectively. Phenol compound of Tawonkong and Socheong cultivars decreased with delay of seeding periods. Radical scavenging activity of Socheong and Jununi cultivars decreased with delay of seeding periods, but Socheong 2 and Socheongja cultivars increased. In the study, phenol compound and radical scavenging activity of small black soybean cultivar were different, depending on cultivars and seeding periods.

Immunohistochemical Expression and Prognostic Value of VEGF, HIF-$1{\alpha}$, EGFR in Non-Small Cell Lung Cancer (비소세포 폐암에서 VEGF, HIF-$1{\alpha}$, EGFR의 면역조직화학적 발현과 예후 인자로서의 역할)

  • Kim, Myung-Sook;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.68 no.1
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    • pp.22-28
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    • 2010
  • Background: Vascular endothelial growth factor (VEGF) is a potent mediator of angiogenesis. VEGF production is regulated by HIF-$1{\alpha}$ and EGFR. This study examined the relationship between the clinicopathological factors and VEGF, HIF-$1{\alpha}$ and EGFR protein overexpression, and evaluated their prognostic value in patients with a surgically resected non-small cell lung cancer (NSCLC). Methods: Patients who underwent a surgical resection at Kangnam St. Mary's hospital were reviewed retrospectively. The core biopsy samples from 54 patients with NSCLC were assembled on a tissue microarray (TMA), and immunohistochemical staining for the VEGF, HIF-$1{\alpha}$ and EGFR proteins was performed. The overexpression of these proteins was evaluated in relation to age, gender, histology and staging by univariate analysis. The clinicopathological prognostic factors were analyzed. Results: Multivariate analysis performed by Cox regression (odds ratio 2.8, 95% CI 1.0~8.2, p=0.046) revealed HIF-$1{\alpha}$ overexpression to be an unfavorable factor. There was no correlation between the overexpression of these proteins and the clinicopathological factors. VEGF showed a positive relationship with EGFR, but there was no statistical significance [$p(x^2)=0.06$]. Conclusion: HIF-$1{\alpha}$ overexpression predicts shorter survival in patients with a surgically resected NSCLC. Therefore, HIF-$1{\alpha}$ may be a poor prognostic factor in NSCLC.

Effects of Soybean Small Peptides on Rumen Fermentation and on Intestinal and Total Tract Digestion of Luxi Yellow Cattle

  • Wang, W.J.;Yang, W.R.;Wang, Y.;Song, E.L.;Liu, X.M.;Wan, F.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.1
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    • pp.72-81
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    • 2013
  • Four Luxi beef cattle ($400{\pm}10$ kg) fitted with ruminal, duodenal and ileal cannulas were used in a $4{\times}4$ Latin square to assess the effects of soybean small peptide (SSP) infusion on rumen fermentation, diet digestion and flow of nutrient in the gastrointestinal tract. The ruminal infusion of SSP was 0 (control), 100, 200 and 300 g/d. Ruminal SSP infusion linearly (p<0.01) and quadratically (p<0.01) increased microbial protein synthesis and rumen ammonia-N concentration. Concentrations of total volatile fatty acid were linearly increased (p = 0.029) by infusion SSP. Rumen samples were obtained for analysis of microbial ecology by real-time PCR. Populations of rumen Butyrivibrio fibrisolvens, Streptococcus bovis, Ciliate protozoa, Ruminococcus flavefaciens, and Prevotella ruminicola were expressed as a proportion of total Rumen bacterial 16S ribosomal deoxyribonucleic acid (rDNA). Butyrivibrio fibrisolvens populations which related to total bacterial 16S rDNA were increased (p<0.05), while Streptococcus bovis populations were linearly (p = 0.049) and quadratically (p = 0.020) decreased by infusion of SSP. Apparent rumen digestibility of DM and NDF were (Q, p<0.05; L, p<0.05) increased with infusion SSP. Total tract digestion of DM, OM and NDF were linearly (p<0.01) and quadratically (p<0.01) increased by infusing SSP. The flow of total amino acids (AA), essential amino acids (EAA) and individual amino acids were linearly (p<0.01) and quadratically (p<0.01) increased with infusion SSP. The digestibility of Lysine was quadratically (p = 0.033) increased and apparent degradability of Arginine was linearly (p = 0.032) and quadratically (p = 0.042) increased with infusion SSP. The results indicated that infusion SSP could improve nutrient digestion, ruminal fermentation and AA availability.