Mulyani, Sri;Setyabudi, Francis.M.C. Sigit;Pranoto, Yudi;Santoso, Umar
한국축산식품학회지
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제37권5호
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pp.708-715
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2017
The acid pretreatment of collagen molecules disrupts their crosslinks and assists in the release of acid-soluble proteins, fats, and other components. Generally, to achieve optimum extraction efficiency, strong acids may be used at a lower acid concentration compared to weak acids. This study aimed to determine the yield and physicochemical properties of gelatins extracted from buffalo hides pretreated with different acids. Hides were extracted with hydrochloric, citric, and acetic acids at concentrations of 0.3, 0.6, 0.9, 1.2, and 1.5 M. A completely randomized design and the least significant difference test were used in the experimental design, and all measurements were performed in triplicate. The highest yield (29.17%) was obtained from pretreatment with 0.9 M HCl. The gel strength did not differ significantly (p>0.05) according to acid type (280.26-259.62 g Bloom), and the highest viscosity was obtained from the 0.6 M citric acid pretreatment. All the gelatins contained ${\alpha}$- and ${\beta}$-chain components and several degraded peptides (24-66 kDa). The color and Fourier-transform infrared spectrum of the gelatin extracted using 0.9 M HCl were similar to those of commercial bovine skin gelatin. In general, the physicochemical properties of the gelatin complied with the industry standard set by the Gelatin Manufacturers Institute of America, revealing that buffalo hide could serve as a potential alternative source of gelatin.
Collagenase existed in the internal organs of filefish Novoden modestrus was isolated with ammonium sulfate and was purified by ion exchange column chromatography with DEAE-Sephadex A-50 and gel filtration with Sephadex G-150. The activity of the purified enzyme was increased 92.4 folds than that of the crude one and the yield of the purified one was $10.9\%$. The optimum conditions showing the maximum activity of the crude enzyme to digest insoluble collagen(Type I) were $55^{\circ}C$ and pH 8.0, while those showing the maximum activity of the purified one were $55^{\circ}C$ and pH 7.75. However, the use of the crude enzyme for skinning of filefish was more profitable because the yield was 800 folds higher than that of the purified one and the cost was also able to economy. When hydrolysis for skinning of filefish was conducted with $0.3\%$(w/w) crude collagenase at $50^{\circ}C$ and pH 8.0 for 3hrs, there was some problem to cause a damage on muscle of the fish by heat. To solve such problem for the skinning, the hydrolysis at $18^{\circ}C$ for 4hrs with $0.3\%$ (w/w) crude enzyme after pretreated with 0.5M acetic acid for 10 min provided a good result for skinning of filefish.
This study was performed to evaluate the efficacy of 0.8% sodium hyaluronic acid solution and crosslinked hyaluronic acid mixture for the prevention of postoperative intraperitoneal adhesion in rats. Forty-five animals were divided into three groups ; 0.9% saline treated control group, 1% sodium carboxymethyl cellulose treated group (SCMC), and 0.8% sodium hyaluronic acid solution and crosslinked hyaluronic acid mixture treated group (SHCH). Adhesions were induced by suturing both the ileal serosa and peritoneum abrased until petechial bleeding occurred. Fourteen days later, adhesions were evaluated clinically and histopathologically. The mean tensile strength was significantly decreased in the SCMC and SHCH groups compared to the control group (p < 0.05), and the SHCH group had the lowest tensile strength. The distance of adhesion site was highest in the control group and significantly decreased in the SHCH group comparing control group (p < 0.05). The inflammatory cell infiltration, collagen hyperplasia and neovascularization of the SCMC and SHCH groups were significantly lower than the control group (p < 0.05). Therefore, it was concluded that the SHCH may be useful to prevent postoperative intraperitoneal adhesion in rats.
This study was carried out to compare the quality properties of chicken breast surimi manufactured by four different procedures/methods. Surimi was made from chicken breast by washing two (T1) or four times (T2) with water as well as by pH adjustments at 3.0 (T3) or 11.0 (T4). The contents of moisture and crude fat were significantly higher in the surimi manufactured from pH-adjusted material than after washing. Again, collagen and yield were significantly higher in chicken breast surimi manufactured from washed than pH-adjusted samples, whereas crude protein was higher in the pH-adjusted than washed surimi samples. There was no significant difference in myofibrillar protein content among the surimi manufactured after different washing times and differences following pH adjustments were found. T4 showed highest myofibrillar protein content rating among the surimi samples. All physical characteristics were higher in pH-adjusted chicken breast surimi than in T1 and T2 washed surimi samples. The pH-adjusted surimi had higher hardness, gumminess and chewiness than washed surimi samples (p<0.05). The chicken breast surimi made by pH adjustments had higher lightness (L*) than when made by washing times, whereas pH 3.0-adjusted surimi samples had lower whiteness (W) then the other surimi samples. Myoglobin content was significantly higher in the surimi manufactured from pH-adjusted chicken breast samples.
Purpose: Facial contouring surgery for improving congenital, acquired deformity and senile change were attempt in past. Recently contouring surgery became more interested subject for improving the flat forehead and temple area. Many synthetic materials were used such as Collagen, silicon, polyacrylamide gel as liquid form and Gore-tex, silicon implant, endotine as solid form. But, these synthetic implants associate complications as foreign body reaction, infection, displacement, granuloma formation and absorption. Auto-fat injection are used for disfigurement of many part of body. We did auto-fat injection for facial contouring of forehead and temple region. Auto-fat injection is suitable without foreign body reaction, displacement, and toxic reaction. Also auto-fat is relatively simple to obtain from patient and less expensive and able to repeat surgeries. Methods: From 2006 to 2009, 150 patients were treated with Auto-fat injection for facial contouring. For follow up, we sent questionnaire to all patients but 110 patients returned answer sheets. The patients consisted of 20 male patients and 90 female patients with an age ranged from 26 to 60, and the mean 43. Fat tissue were injected 6-8 cc in forehead, 7-12 cc in temple area and fat were harvested from thigh and abdomen. Results: In follow up, all patients, showed absorption of injected fat varied degree and except two patients all patients underwent secondary fat injection. Complications were minimal and neuropraxia of facial nerve were recovered. Most of the patients were satisfied with result of procedure, and answered that they recommend same procedure to their friends and will do surgery again. Conclusion: Auto-fat injections were implemented for facial contouring in 150 patients and obtained satisfactory result. Auto-fat injection is relatively easy procedure and applicable widely. Even though, by passing time, some of the injected fats are absorbed, auto-fat injection could be choice of treatment for contouring forehead and temple. With accumulations of cases and development of surgical technique, better result could be expected.
In order to study the implantation mechanism various methods for culture of endometrial cells in vitro have been attempted. However, a disadvantage is that primary cultures of stromal and epithelial cells do not have the ability to differentiate, and therefore cannot be reproduced in the same manner as in vivo endometrium. The object of this study is to establish a three dimensional culture of endometrial cells which are both morphologically and functionally identical to in vivo endometrium. Endometrial tissues obtained after hysterectomies were cut into thin slices and treated with collagenase and trypsin-EDTA. The stromal cells and the epithelial cells were separated by centrifugation and cultured for 24 hours in DMEM media containing 10% FCS, 100 nM progesterone, and 1 nM estradiol. The cultured stromal cells were mixed with collagen gel and solidified, after which it was covered with matrigel. Epithelial cells were inoculated on the top and then cultured for 3 days. The three dimensionally cultured endometrial cells were stained for integrin ${\alpha}1,\;{\alpha}4,\;{\beta}3$, and cyclooxygenase-l, -2 by immunohistochemistry, which all showed strong expression. The cultured epithelial cells showed the formation of microvilli, tight junctions and pinopodes by electron microscopy. Studies are currently under way utilizing this three dimensional culture model to ascertain the interaction between the embryo and human endometrial cells at the time of implantation, and it is thought that further studies into a new culture environment which would allow longer periods of culture will be necessary.
A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.
Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are involved in various pathophysiological processes such as inflammation and carcinogenesis. In a search for inhibitors of COX-2 and iNOS production we found that extracts of Stewartia koreana strongly inhibited NO and $PGE_2$ production in LPS-treated macrophage RAW 264.7 cells. We have now shown that the mRNA and protein levels of iNOS and COX-2 are reduced by the Stewartia koreana extract (SKE). SKE inhibited expression of an NF-${\kappa}B$ reporter gene in response to LPS, and gel mobility shift assays revealed that SKE reduced NF-${\kappa}B$ DNA-binding activity. The extract also inhibited LPS-induced phosphorylation of $I{\kappa}B-{\alpha}$ and nuclear translocation of p65. Administration of the extract reduced the symptoms of arthritis in a collagen-induced arthritic mouse model. These results indicate that Stewartia extracts contain potentially useful agents for preventing and treating inflammatory diseases.
Li, Xiaoguang;Zhang, Qian;Gan, Longzhan;Jiang, Guangyang;Tian, Yongqiang;Shi, Bi
Journal of Microbiology and Biotechnology
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제32권1호
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pp.99-109
/
2022
This study is the first report on production and characterization of the enzyme from an Ornithinibacillus species. A 4.2-fold increase in the extracellular protease (called L9T) production from Ornithinibacillus caprae L9T was achieved through the one-factor-at-a-time approach and response surface methodological optimization. L9T protease exhibited a unique protein band with a mass of 25.9 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This novel protease was active over a range of pH (4-13), temperatures (30-80℃) and salt concentrations (0-220 g/l), with the maximal activity observed at pH 7, 70℃ and 20 g/l NaCl. Proteolytic activity was upgraded in the presence of Ag+, Ca2+ and Sr2+, but was totally suppressed by 5 mM phenylmethylsulfonyl fluoride, which suggests that this enzyme belongs to the serine protease family. L9T protease was resistant to certain common organic solvents and surfactants; particularly, 5 mM Tween 20 and Tween 80 improved the activity by 63 and 15%, respectively. More importantly, L9T protease was found to be effective in dehairing of goatskins, cowhides and rabbit-skins without damaging the collagen fibers. These properties confirm the feasibility of L9T protease in industrial applications, especially in leather processing.
Background: As a kind of common complication of the surgery of perianal diseases, perianal ulcer is known as a nuisance. This study aims to develop a kind of 20(S)-ginsenoside Rg3 (Rg3)-loaded hydrogel to treat perianal ulcers in a rat model. Methods: The copolymers PLGA1600-PEG1000-PLGA1600 were synthesized by ring-opening polymerization process and Rg3-loaded hydrogel was then developed. The perianal ulcer rat model was established to analyze the treatment efficacy of Rg3-loaded hydrogel for ulceration healing for 15 days. The animals were divided into control group, hydrogel group, free Rg3 group, Rg3-loaded hydrogel group, and Lidocaine Gel® group. The residual wound area rate was calculated and the blood concentrations of interleukin-1 (IL-1), interleukin-6 (IL-6), and vascular endothelial growth factor (VEGF) were recorded. Hematoxylin and eosin (H&E) staining, Masson's Trichrome (MT) staining, and tumor necrosis factor α (TNF-α), Ki-67, CD31, ERK1/2, and NF-κB immunohistochemical staining were performed. Results: The biodegradable and biocompatible hydrogel carries a homogenous interactive porous structure with 10 ㎛ pore size and five weeks in vivo degradation time. The loaded Rg3 can be released sustainably. The in vitro cytotoxicity study showed that the hydrogel had no effect on survival rate of murine skin fibroblasts L929. The Rg3-loaded hydrogel can facilitate perianal ulcer healing by inhibiting local and systematic inflammatory responses, swelling the proliferation of nuclear cells, collagen deposition, and vascularization, and activating ERK signal pathway. Conclusion: The Rg3-loaded hydrogel shows the best treatment efficacy of perianal ulcer and may be a candidate for perianal ulcer treatment.
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