• Asian-Australasian Journal of Animal Sciences
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• 제24권4호
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• pp.500-508
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• 2011

Forty-four Gansu Alpine Fine-wool lambs were used to study changes in the activities of three gastric and five pancreatic enzymes under grazing conditions between 0 and 56 days of age. The lambs were slaughtered on days 0, 3, 7, 14, 21, 28, 42 and 56, the abomasal contents, mucosa and pancreas were immediately removed and placed into liquid nitrogen and enzyme activities were determined. Gastric enzyme (chymosin, pepsin and pregastrc esterase) activities were relatively high at birth, especially chymosin, but decreased quickly between day 0 and 21. The activity of pepsin changed insignificantly with increasing age. There was no significant change in the pancreatic enzyme activities (trypsin, chymotrypsin, ${\alpha}$-amylase, lipase and lactase). The activity of trypsin was relatively higher than that of the other pancreatic enzymes, and lactase activity was low. These ontogenic patterns might be under the control of many gut regulatory peptides, the plasma concentrations of which changed simultaneously. Some gastric and pancreatic enzymes were correlated with plasma concentrations of these gut regulatory peptides.

• Journal of mucopolysaccharidosis and rare diseases
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• 제2권1호
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• pp.31-31
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• 2016

Purpose: To describle clinical features and enzyme activity of Vietnamese patients with Mucolipidosis type II. Methods: Clinical features, laboratory and plasma lysosom enzyme activity by 4 MU-Fluorometric assay was studied from 2014-2015 at the Northern referral center of Pediatrics - National Children's Hospital. Results: 16 cases (7 girls and 9 boys) were diagnosed with I-cell bases on clinical symptoms and enzyme activities studies. Diagnosis age was $5.93{\pm}4.28$ years, onset age was recognised from birth to 4 years (median 1.25) with the feature of joint stiffness and bone deformation. All cases presented with the feature of joint stiffness, chest deformation and kyphoscoliosis; Fifteen cases (93.7%) had coarse facial features. No patients had hepatosplenomegaly on abdominal ultrasound, 5/15 patients had heart valves disease. Enzyme assay showed ${\alpha}$-Hexosaminidase of $1,885.9{\pm}338.7$ (nmol/mg plasma/17 hrs), ${\alpha}$-Iduronate sulfatase of $4,534.8{\pm}1,062.9nmol/mg$ plasma/4 hrs). Conclusion: Mucolipidosis II seriously affected the life of the patients with skeletal deformities, contractures develop in all joints and cardiac involvement.

• BMB Reports
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• 제31권4호
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• Applied Biological Chemistry
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• 제40권1호
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• pp.39-42
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• 1997

돼지혈액의 혈장으로부터 ACE 저해제들을 분리 정제하였다. 돼지 도축혈액을 먼저 혈액응고방지제를 첨가한후 원심분리에 의해 혈장과 혈구로 분리한 다음 혈장단백질을 최종농도 4% TCA로 침전시켰다. ACE 저해제들은 TCA 상등액과 혈장단백질 가수분해물로부터 한외여과, 겔여과크로마토그라피, 역상고속액체크로마토그라피 방법으로 분리되었는데 $IC_{50}4값들이 각각 $2\;{\mu}M,\;23\;{\mu}M$로 측정되었다.

• BMB Reports
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• 제28권2호
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• pp.138-142
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• 1995

An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

• 대한의생명과학회지
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• 제16권4호
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• pp.287-292
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• 2010

The purpose of this study was to investigate the effects of soybean peptide on lipid peroxidation, antioxidant enzyme and inflammatory cytokines in male high school taekwondo players. The subjects were divided into 2 groups, which were a soybean peptide intake group (S-peptide, n=13) and a placebo intake group (Placebo, n=10). The s-peptide group took 4 g a day of soybean peptide for 4 weeks. Blood samples were taken from the antecubital vein at before taekwondo performance, after 2 and 4 weeks taekwondo performance. The plasma was analyzed for the antioxidant enzyme activity factor, lipid peroxidation and cytokines. As a result, the s-peptide group decreased lipid peroxidation (MDA) and increased the antioxidant enzyme activity factor (TAS and CAT). The plasma concentration of inflammatory cytokines, IL-6 and TNF-${\alpha}$ were significantly decreased in the s-peptide group after 4 weeks and showed significant differences between the groups. These results indicate that the intake of soybean peptide was positively improved on antioxidant enzyme and inflammatory cytokines in taekwondo player.

• 생명과학회지
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• 제8권6호
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• pp.737-740
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• 1998

멸치액젓으로부터 분리한 혈전용해효소를 rat에 경구투여하여 혈액내에서 효소활성을 확인 하였다. 경구투여 후 0, 1, 2, 3, 4시간 후에 혈액을 채취하여 fibrin분해활성을 본 결과 1시간부터 활성이 나타나기시작하여 3시간째에 가장 높았다. Euglobulin Iysis time(ELT)도 경구투여 2, 3시간후에 활성을 보였다. 또한, euglobulin에 의한 amidolysis acti-vity도 1시간부터 활성이 보이기 시작하여 3시간후에 최고활성에 달했다. 이상의 결과로 동물실험을 통한 혈전용해효소의 경구투여에 의한 활성을 혈액내에서 확인할 수 있었다.

• 한국동물학회지
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• 제34권2호
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• pp.142-147
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• 1991

드렁허리의 혈장에서 얻은 angiotensin I-converting enzyme(ACE)의 활성도를 포유동물과 비교하였다. 드렁허리의 혈장내 ACE는 pH 10에서 가장 높은 활성도를 나타냈으며 포유동물에서 보다 알칼리성이었다. 또한 최적활성도를 나타내는 데 필요한 C1 이온의 요구성은 종에 따라 다르게 나타났다. 드렁허리의 ACE 활성도는 ACE의 활성억제제인 EDTA, teprotide 및 captopril에 의해서 이들의 농도에 따라 억제 되었으며, 이 억제현상은 포유동물의 혈장 ACE와 매우 비슷하였다. 드렁허리의 ACE 활성도는 cobalt에 의하여 증가되었으며, 포유동물에 비해서 열에 불안정하였다. 또한 여러 조직중에서느 뇌에서 가장 높은 ACE 활성도가 측정되었다.

• 대한수의학회지
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• 제32권4호
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• pp.677-681
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• 1992

This experiment was carried out to establish a proper method of indocyanine green(ICG) excrection test for a applicable liver function test in three Korean native cattle average weighing about 450kg and dairy cattle parity of 3~5. The results obtained the half life($T^1/_2$), fractional clearance rate(KICG), retention rate and plasma enzyme activities before or after injection of ICG were as follows. 1. The maximum absorbance of ICG in plasma was at 805nm. 2. Average half life and fractional clearance rate following the injection of ICG 0.25mg/kg body weight were $5.53{\pm}1.27$ minute and $0.131{\pm}0.031$/minute in Korean native cattle, $4.55{\pm}0.68$ minute and $0.156{\pm}0.031$/minute in dairy cattle, respectively. The ICG removal rate was exponentially liner for the first 15 minutes after injection both of Korean native cattle and dairy cattle. 3. Average plasma retention rate when 10, 15, 30 minutes after injection was $35.7{\pm}13.9$, $23.2{\pm}7.1$, $10.8{\pm}3.5%$ in Korean native cattle, $26.8{\pm}3.3$, $14.2{\pm}1.2$, $5.5{\pm}2.2%$ in dairy cattle, respectively. 4. Plasma enzyme activities(AST, ALT, r-GTP) were no variation among the before, during and after injection of ICG. From these results, ICG excretion test to cattle is applicable to evaluation of liver funtion in both clinical and research, and adopted the 15 minutes plasma sample as the sample taken at the ideal time for comparative purposes.

• Animal cells and systems
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• 제8권4호
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• pp.307-312
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• 2004

Fast kinetics of transient pH changes and difference spectrum formation have been investigated following mixing of ADP/ATP with partially purified plasma membrane PM-ATPase of the pathogenic yeast Candida albicans in the presence of five nutrients: glucose, glutamic acid, proline, lysine, and arginine and two analogs of glucose: 2-deoxy D-glucose and xylose. Average $H^+$- absorption to release ratio, indicative of population of ATPase undergoing complete hydrolytic cycle, was found to be 0.27 for control. This ratio varied between 0.25 (proline) to 0.36 (arginine) for all other compounds tested, except for glucose. In the presence of glucose, $H^+$- absorption to release ratio was exceptionally high (0.92). While no UV difference spectrum was observed with ADP, mixing of ATP with ATPase led to a large conformational change. Exposure to different nutrients restricted the magnitude of the conformational change; the analogs of glucose were found to be ineffective. This suppression was maximal in the case of glucose (80%); with other nutrients, the magnitude of suppression ranged from 40-50%. Rate of $H^+$- absorption, which is indicative of E~P complex dissociation, showed positive correlation with suppression of conformational change only in the case of glucose and no other nutrient/analog. Mode of interaction of glucose with plasma membrane $H^+$-ATPase thus appears to be strikingly distinct compared to that of other nutrients/analogs tested. The results obtained lead us to propose a model for explaining glucose stimulation of plasma membrane $H^+$-ATPase activity.