• Korean Journal of Plant Resources
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• 제11권2호
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• pp.195-201
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• 1998

Some morphological character were surveyed and PR-proteinases were induced and characterized from Lilium formosanum Wallace endeimc to Cheju island . Its flower characters were similar to those of white trumpet lilies(Lilium longiflorum Thunb) although its flowering period was later than that of white trumpet lilies and it hadd a wide range of variation among individuals. Six PR-proteinases(II-2, III-1, III-2, IV-1, IV-2 and V) were induced from bulbs by 2.5mM salicylic acid and almost excreted into the intercellular spaces. These PR-proteinases were strongly activated by Ca 2+, , whereas they were strongly inhibited by Cu2+ Co2+ and Fe2+ . Three PR proteinases(II-2, IV-1 and IV-2) were strongly inhibited by 1, 10 -phenanthroline, indicating that these enzymes are metallo-proteinases. Three PR-proteinases(III-1, III-2 and V) had a high sensitivity to PMSF and required $\beta$-mercaptoethanol for their activities. These results indicate that these proteinases are cysteine proteinases.

• Asian-Australasian Journal of Animal Sciences
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• 제33권1호
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• pp.100-110
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• 2020

Objective: The objective of this study was to isolate proteolytic microorganisms and evaluate their effects on proteolysis in total mixed ration (TMR) silages of soybean curd residue. Methods: TMRs were formulated with soybean curd residue, alfalfa or Leymus chinensis hay, corn meal, soybean meal, a vitamin-mineral supplement, and salt in a ratio of 25.0: 40.0:30.0:4.0:0.5:0.5, respectively, on a basis of dry matter. The microbial proteinases during ensiling were characterized, the dominate strains associated with proteolysis were identified, and their enzymatic characterization were evaluated in alfalfa (A-TMR) and Leymus chinensis (L-TMR) TMR silages containing soybean curd residue. Results: Both A-TMR and L-TMR silages were well preserved, with low pH and high lactic acid concentrations. The aerobic bacteria and yeast counts in both TMR silages decreased to about 10⁵ cfu/g fresh matter (FM) and below the detection limit, respectively. The lactic acid bacteria count increased to 10⁹ cfu/g FM. The total microbial proteinases activities reached their maximums during the early ensiling stage and then reduced in both TMR silages with fermentation prolonged. Metalloproteinase was the main proteinase when the total proteinases activities reached their maximums, and when ensiling terminated, metallo and serine proteinases played equally important parts in proteolysis in both TMR silages. Strains in the genera Curtobacterium and Paenibacillus were identified as the most dominant proteolytic bacteria in A-TMR and L-TMR, respectively, and both their proteinases were mainly with metalloproteinase characteristics. In the latter ensiling phase, Enterococcus faecium strains became the major sources of proteolytic enzymes in both TMR silages. Their proteinases were mainly of metallo and serine proteinases classes in this experiment. Conclusion: Proteolytic aerobic bacteria were substituted by proteolytic lactic acid bacteria during ensiling, and the microbial serine and metallo proteinases in these strains played leading roles in proteolysis in TMR silages.

• Journal of agricultural medicine and community health
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• 제22권1호
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• pp.61-74
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• 1997

In case of tissue invading nematode, proteolytic enzyme was required at their parasitic life. Proteinases obtained from these parasites(Toxocara canis, Ansakis spp. and Trichinella spiralis) were extracted, isolated and further purified. And then the analysis for activity and inhibitory effect of proteinases were performed by appropriate substrate. Determination of protein as a circulating antigen was done in use of infected animal serum with above parasites, respectively. For above experimental objects, following procedures were performed. First, enzymatic activity was measured in use of azocasein and inhibitory effect of porteinase were studied by various inhibitors. Second, partially purified proteins containing enzymatic activity were obtained by ion exchange chromatography, ultrafiltration and electrophoretic elution. Third, role of the partially purified protein as a circulating antigen. The results obtained were as follows : 1. Enzymatic activity of each nematode proteinase was varied according to pH. Optimal pH of Toxocara canis, Ansakis spp. and Trichinella spiralis were pH 6.0, pH 5.5 and pH 6.5, respectively. The optimal molarity of buffer was 0.1M phosphate buffer. Although little difference between these proteinases was observed, temperature stability was at least maintained at $4^{\circ}C$ until 5 days. 2. In case of Ansakis spp. and Toxocara canis, enzymatic activity of these proteinases was considerably inhibited by Leupeptin and EDTA. For maximum enzymatic activity of above proteinases, it was required that cysteine residue of enzyme should be protected. And it was suggested that metallo type was contained in enzyme active site. Proteinase of Trichinella spiralis contained metallo type also. 3. Although partial purification was performed in Ansakis spp. and Toxocara canis, proteins maintaining enzymatic activity were identified as a circulating antigen. From SDS-PAGE and immunoblot, 25 kDa was presented in Ansakis spp.. Specific antigen of Toxocara cains was 110 kDa protein fraction. 55 and 42 kDa proteins were reacted with normal serum. Trichinella spiralis 60 kDa protein fraction was successfully purified from excretory materials in culture. As a result of immune-reaction with Trichinella spiralis infected serum, highly purified 60 kDa protein was maintained antigenicity until final purification step.

• The Korean Journal of Food And Nutrition
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• 제30권2호
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• pp.211-217
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• 2017

In the present study, we investigated the protective effect of various grain methanolic extracts against UVB-induced photo-aging in human skin fibroblasts. Various grain methanolic extracts were evaluated for their antioxidant compounds and activities. 2,2-Ddiphenyl-1-picryhydrazyl radical (DPPH) and ABTS 2,2-azino-bris-(3-ethylbenzoth iazoline-6-sulphonic acid) radical cation scavenging activities have been used to measure the relative antioxidant activities of extracts from grains. The content of total polyphenolics in the extracts were evaluated using spectrophotometric methods. Human skin fibroblast (Hs68) cells were pretreated with various grain methanolic extracts ($25{\mu}g/mL$). Skin toxicity was simulated by exposing the cells to UVB ($30mJ/cm^2$) irradiation. In response to the UVB-irradiation, an increased amount of matrix metallo-proteinases (MMPs) release was observed, whereas pretreatment of various grain methanolic extracts significantly inhibited the production of MMP-1 in Hs68 cells. We also found that pretreatment of the extracts significantly decreased UVB-induced reactive oxygen species and significantly increased total collagen content in Hs68 cells. These results provide that grains could be regarded as a potential ingredient in natural cosmetics, used for UVB protection.

• Journal of the Society of Cosmetic Scientists of Korea
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• 제32권2호
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• pp.117-121
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• 2006

Although many studies have been performed to elucidate the molecular consequence of factors that regulate skin aging, little is known about the effect of green tea catechins except EGCG. The matrix metalloproteinase (MMP), can degrade matrix proteins and results in a collagen deficiency in photodamaged skin, are known to play an important role in photoaging. This study, investigated the effects of green tea catechins on the UVA-induced MMP-1 expression, activity of MMP-2 and synthesis of type I procollagen in human dermal fibroblasts. We examined eight catechins that naturally exist in green tea leaves and compared their efficacies among them. Most of catechins inhibited the expression of MMP-1 in dose dependent manner, and the levels were reduced, especially, 57.4 and 68.2% by treatment with $1{\mu}M$ of epigallocatechin-3-gallate (EGCG) and gallocatechin-3-gallate (GCG), respectively. Also, catechins significantly suppressed the activities of MMP-2. Catechins also induced the expression of type I procollagen, however, they acted only at the concentration below $1{\mu}M$ interestingly. Furthermore, when EGCG:GCG:ECG had the ratio of 0.5:1.5:.1.3, they presented the most effective on procollagen synthesis. Therefore, we concluded that catechins significantly inhibited MMPs and induced collagen synthesis. Taken together, all these results suggested that green tea catechins might be good natural materials act as an anti-photoaging and a skin-aging improving agent.

• Journal of Veterinary Science
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• 제24권6호
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• pp.83.1-83.12
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• 2023

Background: Ellipticine (Ellip.) was recently reported to have beneficial effects on the differentiation of adipose-derived stem cells into mature chondrocyte-like cells. On the other hand, no practical results have been derived from the transplantation of bone marrow stem cells (BMSCs) in a rabbit osteoarthritis (OA) model. Objectives: This study examined whether autologous BMSCs incubated with ellipticine (Ellip.+BMSCs) could regenerate articular cartilage in rabbit OA, a model similar to degenerative arthritis in human beings. Methods: A portion of rabbit articular cartilage was surgically removed, and Ellip.+BMSCs were transplanted into the lesion area. After two and four weeks of treatment, the serum levels of proinflammatory cytokines, i.e., tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), were analyzed, while macroscopic and micro-computed tomography (CT) evaluations were conducted to determine the intensity of cartilage degeneration. Furthermore, immuno-blotting was performed to evaluate the mitogen-activated protein kinases, PI3K/Akt, and nuclear factor-κB (NF-κB) signaling in rabbit OA models. Histological staining was used to confirm the change in the pattern of collagen and proteoglycan in the articular cartilage matrix. Results: The transplantation of Ellip.+BMSCs elicited a chondroprotective effect by reducing the inflammatory factors (TNF-α, PGE2) in a time-dependent manner. Macroscopic observations, micro-CT, and histological staining revealed articular cartilage regeneration with the downregulation of matrix-metallo proteinases (MMPs), preventing articular cartilage degradation. Furthermore, histological observations confirmed a significant boost in the production of chondrocytes, collagen, and proteoglycan compared to the control group. Western blotting data revealed the downregulation of the p38, PI3K-Akt, and NF-κB inflammatory pathways to attenuate inflammation. Conclusions: The transplantation of Ellip.+BMSCs normalized the OA condition by boosting the recovery of degenerated articular cartilage and inhibiting the catabolic signaling pathway.

• Journal of Acupuncture Research
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• 제19권2호
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• pp.51-64
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• 2002

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.