• Title/Summary/Keyword: trypsin activity

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Development of Experimental Animal Model with Duodenal Ulcer for Nutritional Study and its Nutritional Characteristics (영양실험을 위한십이지장 궤양 흰쥐 실험 모델 개발과 그 영양생리적특성)

  • 김창임;이연숙;최혜미
    • Korean Journal of Community Nutrition
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    • v.1 no.2
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    • pp.260-268
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    • 1996
  • This study aimed to develop an experimental animal model of duodenal ulcer for application to nutrition, and to verify the nutritional characteristics. The 7 different doses and 8 different injections of cysteamine and 2 kinds of diets with different casein levels(10%, 20%) were tested to examine the incidence and the process of duodenal ulcer in female Sprague-Dawley rats. The result showed that the duodenal ulcer was induced when rats fed 10% casein diet(for 9 days) were injected 6 times with 13mg/100g BW cysteamine. The first injection was conducted after 24 hours of fasting, followed by the 2nd shot after 3 hours. The following shots were repeated every 3 day(4th and 7th days). Duodenal ulcer was observed 3 days after the last injection by optical microscope and the naked eye. The characteristics of nitrogen bioavailability of duodenal ulcer model were as followings : 1) gastric emptying rate was faster, 2) trypsin activity at duodenum was higher, 3) total protein concentration at serum and nitrogen retention rate were lower than the control. (Korean J Community Nutrition 1(2) : 260-268, 1996)

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Effect of Retinoic Acid on Proliferation and Differentiation of Preadipocytes from Male and Female Pigs

  • Song, Mi-Yeon;Dang, Chang-Gwon;Chung, Chung-Soo
    • Journal of Animal Science and Technology
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    • v.53 no.3
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    • pp.223-226
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    • 2011
  • The current study was undertaken to determine the effect of retinoic acid (RA) on proliferation and differentiation of preadipocytes from male and female pigs. The preadipocytes were isolated from new-born male and female pigs by collagenase digestion and washed three times one day after seeding (designated as day 0 of culture). RA was included in the media at various concentratives from day 0 to 2. The cell number was measured on day 2 with hematocytometer after trypsin digestion. Cell differentiation was determined on day 6 by measuring glycerol-3-phosphate dehydrogenase activity. RA (0.1, 1 and 10 uM) showed no effect on proliferation of preadipocytes from both male and female pigs. However, RA significantly decreased differentiation of pig preadipocytes. Degree of differentiation with 0.1 uM, 1 uM and 10 uM of RA treatment was 80%, 41% and 29% respectively, compared with control. Similar inhibitory effect was found in the female pigs; 77%, 28% and 16% respectively. It is interesting that RA treated on cell proliferation stage had no effect on proliferation but had a strong inhibitory effect on differentiation which is happening in the late stage of cell culture.

Evaluation of Cigarette Quality by Use of ??1-Protease Inhibitor (단백질 분해효소억제제를 이용한 담배의 품질평가)

  • 손형옥;임흥빈;이영구;이동욱;김용태
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.27-33
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    • 1991
  • Current studies indicated that emphysema in smokers might be due, in part, to the local suppression of G, -protease inhibitor(u, -Pl) in lung by reactive oxygen species in cigarette smoke or smoke-activated lung neutrophiles. In the present works, we examined the possibility that a measure which inactivated $\alpha$l-Pl by cigarette smoke could be an alternative method to evaluate the cigarette quality, In order to determine the inactivation of $\alpha$1, -Pl, trypsin inhibitory capacity(TIC) was assayed. A rapid loss of $\alpha$1, -Pl activity occurred when $\alpha$1-Pl solutions was exposed the gas phase or total particulate matter(TPM) obtained from various brands. The inactivation of $\alpha$1-Pl by gas phase was dependent upon the number of puffs and the age of the smoke. However, that by TPM was rather decreased since 2 puffs and also showed no more change over 24hrs after exposing. Inactivation of $\alpha$1-Pl determined by our suggested method(5 puffs, 24hours of aging after exposing) using various commercial cigarettes exhibited that high tar brands has inactivated it more strongly than low tar cigarettes. But the ability of some brands to inactivate $\alpha$1-Pl does not correlate with the content of tar or nicotine. These results so여esc that the degree of $\alpha$1-Pl inactivation by cigarette smoke may be a useful index for the evaluation of cigarette quality and that it should be also contribute to the manufacture of less hazardous cigarettes.

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Experimental Study of Chungganhaeju-tang (Qingganjiejiu-tang) on Oxidative Stress (청간해주탕(淸肝解酒湯)의 항산화 작용에 관한 실험적 연구)

  • Lee, Ji-Eun;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.32 no.2
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    • pp.188-202
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    • 2011
  • Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Chungganhaeju-tang (Qingganjiejiu-tang, CGHJT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the effects of Chungganhaeju-tang on oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of CGHJT on DPPH (1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase, and measured cell viability, and proliferation. In the cell culture model, we measured the activities of superoxide dismutase (SOD), and catalase (CAT) after CGHJT treatment in C34 and E47 cell lines, HepG2 cells transfected with/without the cytochrome P450 2E1 (CYP2E1) gene. In vivo, we measured malondialdehyde levels in the liver tissue and alcohol concentration in the blood. Results : CGHJT showed significant free radical scavenging activity against DPPH and xanthine oxidase in the in vitro study, and increased cell viability, proliferation, and activities of superoxide dismutase, catalase in C34 and in E47 cell lines. CGHJT reduced malondialdehyde levels and blood alcohol concentration in vivo, as well. Conclusions : This study suggests that CGHJT has antioxidant effects on oxidative stress by reducing lipid peroxidation and inhibiting the ethanol induced suppression of antioxidant enzyme activities.

Purification and Characterization of a Novel Extracellular Alkaline Phytase from Aeromonas sp.

  • SEO MYUNG-JI;KIM JEONG-NYEO;CHO EUN-AH;PARK HOON;CHOI HAK-JONG;PYUN YU-RYANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.745-748
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    • 2005
  • A phytase from Aeromonas sp. LIK 1-5 was partially purified by ammonium sulfate precipitation and DEAE-Sephacel column chromatography. Its molecular weight was 44 kDa according to SDS-PAGE gel. Enzyme activity was optimal at pH 7 and at $50^{\circ}C$. The purified enzyme was strongly inhibited by 2 mM EDTA, $Zn^{2+},\;Co^{2+},\;or\;Mn^{2+}$, and activated by 2 mM $Ca^{2+}$. The K_m value for sodium phytate was 0.23 mM, and the enzyme was resistant to trypsin. The N-terminal amino acid sequence of the phytase was similar to that of other known alkaline phytases. The phytase was specific for ATP and sodium phytate, which is different from other known alkaline phytases. Based on the substrate specificity, the phytase may therefore be a novel alkaline phytase.

Characteristics of Peptide Assimilation by Helicobacter pylori: Evidence for Involvement of Cell Surface Peptidase

  • YUN SOON-KYU;CHOI KYUNG-MIN;UHM CHANG-SUB;PARK JEONG-KYU;HWANG SE-YOUNG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.899-902
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    • 2005
  • Peptide assimilation by Helicobacter pylori was investigated using L-phenylalanyl-3-thia-phenylalanine (PSP) as a detector peptide; the release of thiophenol upon enzymatic hydrolysis of PSP was spectrophotometrically detected with the aid of 5,5'-dithiobis[2-nitrobenzoic acid] (DTNB). By adding PSP to whole-cell suspension, thiophenol was produced progressively, resembling that found in Esherichia coli or Staphylococcus aureus. Interestingly, the rate of thiophenol production by H pylori in particular was markedly reduced when cells were pretreated with trypsin, indicating surface exhibition of peptidase. According to the competitive spectrophotometry using alanyl-peptides, H pylori did not appear to assimilate PSP through the peptide transport system. No discernible PSP assimilation could be ascertained in H pylori cells, unless provided with some additives necessary for peptidase activity, such as $Ni^{2+}\;or\;Mg^{2+}$ and an appropriate concentration of potassium or ammonium salts. These observations strongly suggest that, regardless of a presumptive peptide transport system, peptide assimilation of H. plori appears to be highly dependent upon milieu conditions, due to unique peptidase exhibition on the cell surface.

Antioxidative Activities of Hydrolysates from Duck Egg White Using Enzymatic Hydrolysis

  • Chen, Yi-Chao;Chang, Hsi-Shan;Wang, Cheng-Taung;Cheng, Fu-Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.11
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    • pp.1587-1593
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    • 2009
  • Duck egg white (DEW) hydrolysates were prepared by five enzymes (papain, trypsin, chymotrypsin, alcalase, and flavourzyme) and their antioxidant activities investigated in this study. DEW hydrolyzed with papain (DEWHP) had the highest peptide content among the five enzymatic treatments. Besides, the peptide content of DEWHP increased when the enzyme to substrate ratio (E/S ratio) increased. It was suggested that higher E/S ratio contributed to elevate the degree of hydrolysis of DEW effectively. Similar results were also obtained by Tricine-SDS-PAGE. In addition, SDS-PAGE patterns indicated papain was the only one amongst all enzymes with the ability to hydrolyze DEW. In antioxidant properties, DEWHP showed more than 70% of inhibitory activity on linoleic acid peroxidation and superoxide anion scavenging. Moreover, the $Fe^{2+}$ chelating effect of DEWHP was greater than 90%, while no significant difference was observed in DPPH radical scavenging and reducing ability. The results of peptide contents, antioxidant activities and electrophoresis suggested that the higher the peptide content, the stronger the antioxidant activities in DEWHP.

Altered Gene Expression in Cerulein-Stimulated Pancreatic Acinar Cells: Pathologic Mechanism of Acute Pancreatitis

  • Yu, Ji-Hoon;Lim, Joo-Weon;Kim, Hye-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.409-416
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    • 2009
  • Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with $10^{-8}$ M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.

Anti-rheumatoidal Effect of Sulfuretin Isolated from the Heartwood of Rhus veniciflua in Rats and Mice

  • Choi, Jong-Won;Yoon, Byung-Jae;Keun-Huh;Park, Kun-Young;Lee, Kyung-Tae;Park, Hee-Juhn
    • Preventive Nutrition and Food Science
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    • v.7 no.4
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    • pp.347-352
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    • 2002
  • The present study was undertaken to evaluate the anti-rheumatoidal arthritis effect of the R. verniciflua heartwood extract, its EtOAc fraction, and its primary flavonoids, sulfuretin and fustin. All test samples showed variably significant inhibitory effects on hind paw edema and trypsin inhibitor activity induced by Freund's complete adjuvant reagent (FCA reagent), and on vascular permeability caused by acetic acid. Treatment with 10 mg/kg (i.p.) sulfuretin for seven days inhibited edema formation by 54.2$\pm$3.0%. Test samples, especially sulfuretin, shifted the values of biochemical parameters such as serum-cholesterol, serum-triglyceride and serum-total protein toward the normal and restored the numbers of leucocytes and platelets. These results suggest that the heartwood of R. verniciflua reduces immunological injuries caused by FCA reagent provides evidence that suluretin is an active anti-rheumatoid arthritis agent.

Purification and Characterization of a Collagenase from the Mackerel, Scomber japonicus

  • Park, Pyo-Jam;Lee, Sang-Hoon;Byun, Hee-Guk;Kim, Soo-Hyun;Kim, Se-Kwon
    • BMB Reports
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    • v.35 no.6
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    • pp.576-582
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    • 2002
  • Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.