• Applied Microscopy
• /
• v.37 no.2
• /
• pp.93-102
• /
• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.4
• /
• pp.509-513
• /
• 1999

The molecular size reduction and the formation of bitterness during a tryptic hydrolysis of soybean 11S glycinin were determined by using quantitative analysis and organoleptic evaluation. The 11S glycinin of 90% purity was prepared by cryoprecipitation and Con A Sepharose 4B affinity chromatography, and hydrolyzed with trypsin in a pH-stat reactor for 4 h. Bitterness was formed within 1 h of hydrolysis, and then slowly increased up to $3.5\times10^{-5}$ M quinine-HCl equivalent. The extent of hydrolysis (DH) was 7% at 1 h and increased up to 12% by the end of the reaction. The -amino nitrogen content increased from an initial 0.7 mM to 7 mM at the end of the period. The SDS-PAGE analysis showed that the acidic subunit of 11S glycinin was mostly hydrolyzed. The GP-HPLC analysis indicated that the bitterness was mainly contributed by the peptide fractions of molecular weights of 360-2,100 Da.

• Journal of Nutrition and Health
• /
• v.3 no.3
• /
• pp.149-159
• /
• 1970

This study is to investigate the status of anemia, especially iron deficiency anemia among pre-school children in rural area in Korea. The survey was conducted in Sang-dae Ri, Yusong Myon, Daedok Gun, Chung Chong Nam-Do from July 30 th to August 12th, 1968. The measurements were done of height, weight, hematologist and biochemical levels on ninety-two pre-school children, 47 male, and 45 female, one to six years of age. Hemoglobin was determined by the method of cyanmethemoglobin and hematocrit by micro hematocrit centrifuge. The determination of serum iron, iron-binding capacity was done by the method of Ramsay using bathophenanthroline and the serum albumin was determined by Biuret Reaction. The results of this study are as follows: 1) 54.4 percent of the pre-school children weighed less than 90 percent of the Korean General Standard Weight level. 2) The average hemoglobin level was $11.0{\pm}1.57gm/100ml$, 38.0 percent of the children were anemic with less than 1.0gm/100ml. Of the anemic children 60 percent were below the Korean General Standard Weight level. 3) 27.5 percent of the pre-school children were found to have below 32 percent of a hematocrit values and 28.0 percent showed less than 33 percent in M.C.H.C. These results showed that the incidence of hypochromic anemia in these pre-school children was high. 4) 37.9 percent of these children had a serum iron level less than $50{\mu}g/100ml\;and\;31.0\;percent\;had\;a\;TIBC\;above\;400{\mu}g$ while 48.3 percent showed a transferrin saturation lower than 15 percent. On the basis of these findings, it is concluded than the cause of this anemia was iron deficiency. 5) In this group there was a little evidence of low total serum protein levels. However, 10.4 percent of the children had a deficient serum albumin level, below 2.80 gm/100ml while 51.7 percent had a low level, less than 3.50gm/100ml, and 34.5 percent of the children had a low level of TIBC, less than $350{\mu}g/100ml$, and considering these facts, it is suggested that some of the anemias have a multiple causes through protein deficiency and repeated chronic infection apart from iron deficiency.

• Journal of Microbiology
• /
• v.45 no.5
• /
• pp.402-408
• /
• 2007

A strain producing a potent protease was isolated from turban shell. The strain was identified as Bacillus sp. S17110 based on phylogenetic analysis. The enzyme was purified from culture supernatant of Bacillus sp. S17110 to homogeneity by ammonium sulfate precipitation, SP-Sepharose, and DEAE-Sepharose anion exchange chromatography. Protease activity of the purified protein against casein was found to be stable at pH 7 to pH 10 and around $50^{\circ}C$. Approximately 70% of proteolytic activity of the enzyme was detected either in the presence of 100 mM SDS or Tween 20. The enzyme activity was enhanced in the presence of $Ca^{2+},\;Zn^{2+},\;Mg^{2+}$, but was inhibited by EDTA, indicating that it requires metal for its activity. The purified enzyme was found to be a monomeric protein with a molecular mass of 75 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. The purified enzyme was analyzed through peptide fingerprint mass spectra generated from matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and a BLAST search, and identified as immune inhibitor A (inhA) deduced from nucleotide sequence of B. cereus G9241. Since InhA was identified as protease that cleave antibacterial proteins found in insect, inhA-like protease purified from Bacillus sp. S17110 might be pathogenic to sea invertebrates.

• Microbiology and Biotechnology Letters
• /
• v.44 no.4
• /
• pp.442-451
• /
• 2016

The aim of this study was to investigate the anti-inflammatory effect of Sargassum miyabei Yendo ethanol extract (SMYEE) using RAW 264.7 cells and croton oil-induced Balb/c mice. SMYEE inhibited the production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and $IL-1{\beta}$] and nitric oxide in lipopolysaccharide (LPS)-induced inflammatory response. In addition, SMYEE suppressed the expression of inducible nitric oxide, cyclooxygenase-2, and nuclear factor-kappa B. Further, SMYEE inhibited the expression of mitogen-activated protein kinases (MAPKs), such as extra cellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase. In ear edema test, edema formation in the SMYEE treatment was lower than that in the positive control and was similar to that in the prednisolone treatment group. Photomicrographs of mice ear tissue showed a reduction in dermal thickness and number of infiltrated mast cells. Therefore, our results indicate that SMYEE exerts an anti-inflammatory effect via inhibition of nuclear factor ${NF}-{\kappa}B$ and MAPK activation and can be used as a natural source of anti-inflammatory compounds.

• Microbiology and Biotechnology Letters
• /
• v.9 no.4
• /
• pp.185-190
• /
• 1981

1. $\alpha$-amylase from B. circulans F-2 was purified with specific activity 55.0 u/mg. protein (about 23 times of the original specific activity) and the yield of 25.5%, by means of corn starch absorption, salting out with ammonium sulfate (80% saturation), gel filtration on Bio-Gel P-100 and DE-32 column chromatography. 2 The purified enzyme showed two closely migrated protin bands on polyacrylamide disc gel electrophoresis, both of which have amylase activity judging from the activity staining of the gel. On SDS-polyacrylamide disc gel electrophoresis, however, the purified enzyme showed a single band suggesting that those two bands are the charge isomers of an amlyase having the slightly different charge. 3. Plot of log mobility of two bands versus polyacrylamide gel concentration according to Hedrick and Smith gave the parallel lines indicating them to be charge isomers. 4. To confirm the action pattern of two enzyme protein bands, each band was separated and was eluted from the gel and eluates were incubated with soluble starch. Oligosaccharide pattern produced by each eluate was examined by paper chromatography. The eluates of two bands showed the same action pattern. 5. The maltohexaose was the only hydrolysis product of soluble starch in the early stage of hydrolysis.

• YAKHAK HOEJI
• /
• v.48 no.6
• /
• pp.323-327
• /
• 2004

Whitening effect, which decreases the skin pigmentation, is the one of important targets in cosmetics. This study was investigated the effects of Scirpi rhizoma on ant ioxidation and melanogenesis. S.rhizoma is a rhizome of Scirpus fluviatilis G. a perennial Cyperaceae species of wide occurrence in Asia, Europe, Africa and North America. S.rhizoma shown scavenging activities of free radicals and reactive oxygen species (ROS) with the IC50 of 638${\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 21.7${\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S.rhizoma treatment (48 h) suppressed the biosynthesis of melanin up to 27% and reduced tyrosinase activity up to 31% at 100${\mu}g/ml$ in B16 melanoma cells. S.rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expres- sion in protein level. These results suggest that S.rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore S.rhizoma may be useful as new whitening agent due to the antioxidant effect and the inhibitory effect against melanogenesis.

• Journal of Embryo Transfer
• /
• v.21 no.1
• /
• pp.45-51
• /
• 2006

This study was carried out to investigate effect of claw trimming on milk yield and its composition in Holstein at different lactation stages. 1 . There was no difference in daily milk yield between control and claw trimming in early, mid and late lactating Holsteins. 2. Somatic cell count (SCC) was lower in early lactation and it was higher in late lactation when claws were trimmed in Holstein. However, claw trimming did not affect SCC during mid lactation in Holstein. 3. Milk fat, protein and total solids were decreased during late lactation in Holstein after claw trimming. However, milk composition was not affected by claw trimming in early and mid lactating Holsteins.

• Asian-Australasian Journal of Animal Sciences
• /
• v.7 no.1
• /
• pp.69-74
• /
• 1994

The relationship of thyroid status and certain blood biochemical constituents with body weight gains (ADG) and age (13 to 96 weeks) was studied in Holstein Friesian ${\times}$ Hariana ($1/2F{\times}1/2H$) crossbred male calves by assessing their plasma triiodo thyronine ($T_3$), Thyroxine ($T_4$), sodium, potassium, total proteins and cholesterol level at two energy levels. Body weight gains (ADG) were higher during the 50 to 72 weeks of age and declined thereafter, the plasma $T_3$ conc. was aignificantly (p < 0.01) higher during this period compared to all other periods. There was no significant variation due to energy level. Overall mean plasma $T_3$ conc. was $1.19{\pm}0.12ng/ml$. Plasma $T_4$ conc. did not show any significant variation either between the different age periods or between the two energy levels. The mean plasma $T_4$ conc. was $37.34{\pm}1.32ng/ml$. The plasma sodium and potassium concentration did not vary significantly due to energy levels. But amongst the different age periods, sodium concentration was highest ($147.70{\pm}2.29mEq/L$) during 49-60 weeks of age and lowest ($134.70{\pm}1.78mEq/L$) during 13-24 weeks, where as for potassium concentration changes were nonsignificant. There was very little variation amongst other periods. Plasma protein level was higher at 100% energy level than at 75%. Amongst the different age period, it was significantly lower ($6.44{\pm}0.36$) during 13 to 24 weeks of age than at 37 to 48 weeks of age ($7.14{\pm}0.11$). Plasma cholesterol values were higher for 75% energy level than that of 100%. Between the periods it was highest during 61 to 72 weeks of age and the difference amongst the age period were highly significant.

• Korean Journal of Animal Reproduction
• /
• v.24 no.4
• /
• pp.429-437
• /
• 2000

This study investigated the successful introduction of genes of erythropoietin (EPO) and enhanced green fluorescent protein (EGFP) in bovine embryos following nuclear transfer of bovine fetal fibroblasts (bFF), which were transfected by retrovirus vector system. Non-starved bFF were, transferred into perivitelline space of enucleated oocytes. The bFF-oocyte units were accomplished by cell to cell fusion and activated with calcium inophore and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CRlaa medium for 8 days. Out of 187 (EPO) and 210 (EGFP) bovine eggs reconstructed by nuclear transfer, 149 (EPO : 80.0%) and 158 (EGFP : 75.2%) embryos were cleaved, and among them 36 (EPO : 24.2%) and 35 (EGFP : 22.2%) embryos developed to the blastocyst stage. Of these blastocysts, 100% integration of EPO gene in 36 embryos was determined by PCR, and 100% expression of EGFP gene in 35 embryos was observed under the fluorescent microscope. This result indicates that bovine oocytes reconstructed by nuclear transfer of transfected bFF can successfully develop to the blastocyst stage. Furthermore, this novel procedure may be presumably an attractive method efficiently to produce the transgenic cattles.