• Asian-Australasian Journal of Animal Sciences
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• 제31권1호
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• pp.129-137
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• 2018

Objective: The objective of this research was to evaluate the physico-chemical, microbiological and sensorial qualities of restructured steaks processed from beef trimmings (grade I and II) and frozen beef (fresh beef as control and frozen beef). Methods: Beef trimmings from commercial butcher were collected, designated into 4 treatments differing in beef trimmings grade and freezing, processed into restructured steaks with 1% microbial transglutaminase and then analyzed for product quality. Results: The results showed that all meat from different groups could be tightly bound together via cross-linking of myosin heavy chain and actin as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Microbial counts of psychrotrophic and mesophilic bacteria were not affected by treatments (p>0.05), and no detectable of thermophilic bacteria were found. Regarding effect of beef trimmings grade, steaks made from beef trimmings grade II (16.03% fat) showed some superior sensorial qualities including higher tenderness score (p<0.05) and tendency for higher scores of juiciness and overall acceptability (p<0.07) than those made from beef trimmings grade I (2.15% fat). Moreover, a hardness value from texture profile analysis was lower in steaks processed from beef trimmings grade II than those made from grade I (p<0.05). Although some inferior qualities in terms of cooking loss and discoloration after cooking were higher in steaks made from beef trimmings grade II than those made from beef trimmings grade I (p<0.05), these differences did not affect the sensory evaluation. Frozen beef improved the soft texture and resulted in effective meat binding as considered by higher cohesiveness and springiness of the raw restructured product as compared to fresh beef (p<0.05). Conclusion: The results indicated the most suitable raw beef for producing restructured steaks without detrimental effect on product quality was beef trimmings grade II containing up to 17% fat which positively affected the sensory quality and that frozen beef trimmings increased tenderness and meat binding of restructured beef steaks.

• Journal of Oral Medicine and Pain
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• 제21권2호
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• pp.331-349
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• 1996

Cytochrome P45O is an oxidase involved in oxidation of alcohol and is known to be an activator of carcinogen. The present study was performed to study the effect of alcohol and cold stress on the expression of Cytochrome P450 IIEl (CYPIIE1) In the liver and salivary glands in rats by an immunoblot analysis. Sixteen rats were divided into 4 groups; 1)rats belonging to group I were allowed to take 15%(v/v) ethyl alcohol as a drink ad libitum: 2)rats of group II were bathed in cold water for 30 sec twice a day (during the one-week experiment); 3)rats comprising group III were received alcohol and cold stress as described above; 4)rats of group IV were selected as a control. The rat were sacrificed at the end of the one-week experiment. The livers and parotid and submandibular salivary glands were removed and stored at -2$0^{\circ}C$ until use. The stored organs were homogenized for 10 sec and the supernatants were obtained by centrifugation. The proteins of the supernatants were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to Western blotting. The blotted membranes were incubated with polyclonal antibodies to CYPIIEI . The obtained results were as follows : 1. The expression of CYPIIEl was apparently negative in the liver and salivary glands of group IV, wheras its expression was marked in the experiment groups I, II. and III. 2. No difference in the expression of CYPIIEl in the liver and salivary glands was observed between the experiment groups I, II, and III. 3. Among the experiment groups, the expression of CYPIIE1 in the liver was much greater than in the salivary glands. The expression of CYPIIE1 in the submandibular gland was weakly positive but was greater than in the carotid gland.

• Restorative Dentistry and Endodontics
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• 제19권2호
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• pp.485-498
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• 1994

Porphyromonas endodontalis is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections and this microorganism possesses a potential for pathogenicity. The purpose of this study was to compare the membrane components of Porphyromonas endodontalis and Porphyromonas gingivalis and to study the immune reaction patterns of Porphyromonas endodontalis with patients with periapical lesion. Porphyromonas endodontalis (ATCC 35406), Porphyromonas gingivals serotypea (381), serotype b(W50), serotype c(A7A1-28) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells and human sera were obtained from patients and dental students. Indirect immunofluorescence method was used to study on the cross reaction between Porphyromonas endodontalis and Porphyromonas gingivalis serotype a, b, c antigen. Total membrane protein profiles of Porphyromonas endodontalis antigen were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the reactivity of antigenic components of Porphyromonas endodontalis against sera of patients and rabbit anti-Porphyromonas endodontalis antisera were assessed by Immunoblotting method. The following results were obtained : 1. Antigens of Porphyromonas endodontalis has multiple antigenic components, and both patients with periapical lesion and normal healthy individual showed immune response to this. 2. Patients group and healthy individual group showed a diversity of immune reaction pattern but they showed immune response against 43kd protein. 3. Patients with periapical lesion showed more diverse immune response than healthy individual and in some patients, much more bands appeared to lower molecular weight protein. 4. According to indirect immunofluorescence and Immunoblotting study, Porphyromonas endodontalis did not share common antigen with Porphyromonas gingivalis serotype a, b, c.

• 대한약침학회지
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• 제17권1호
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• pp.44-50
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• 2014

Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as $Fe^{2+}$ and $Hg^{2+}$ inhibited the fibrin hydrolysis completely, but $Zn^{2+}$ enhanced it. FE-32kDa hydrolyzed ${\alpha}$-chain but did not hydrolyze ${\beta}$-chain and ${\gamma}$-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of $2{\mu}g$. Conclusions: FE-32kDa is a ${\alpha}$-fibrin(ogen)olytic metalloprotease that requires $Zn^{2+}$ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture.

• 대한의용생체공학회:의공학회지
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• 제41권3호
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• pp.138-145
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• 2020

Adipocytes are the main constituent of adipose tissue. Understanding the molecular basis of adipogenesis is pivotal to finding the therapeutic targets for treatment of obesity. Melatonin is associated with obesity and its mechanism is currently under intensive investigation. The objective of this study was to investigate the effect of melatonin on adipogenesis in differentiating preadipocytes. 3T3-L1 preadipocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5% calf serum at 37℃ with 5% CO₂ in a humidified incubator. Differentiation was induced using DMEM with 10% fetal bovine serum supplemented with MDI two days after cell confluence (day 0). Cells were treated with 0, 10 and 100 μM melatonin on either day 0 or day 5. 72 hours after each treatment, lipid accumulation was measured by oil red O staining. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to membranes. As a result, lipid accumulation decreased with melatonin treatment. ERK pathway, activated when differentiation is induced, also decreased with an increase in melatonin concentration. Furthermore, the expression of key adipogenic factors, C/EBPα, C/EBPβ, and PPARγ, were reduced by melatonin treatment. These results imply that melatonin may inhibit the process of adipogenesis and may have a role as a new anti-obesity agent.

• 한국축산식품학회지
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• 제32권6호
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• 미생물학회지
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• 제45권3호
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• pp.257-262
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• 2009

Bacillus licheniformis WL-12의 carboxymethyl cellulase (cellulase) 유전자를 함유한 대장균 균체 파쇄상등액으로부터 DEAE-Sepharose와 Q-Sepharose 컬럼 크로마토그래피를 통해 cellulase를 정제하였다. 정제된 효소의 비활성은 163 U/mg이었으며, SDS-PAGE에 의해 측정된 분자량은 약 49.5 kDa으로 나타났다. pH 5.5와 $55^{\circ}C$에서 최대 반응활성을 보였으며, SDS (5mM)에 의해서는 cellulase의 활성이 완전히 저해되었고 $Cu^{2+}$5mM)에 의해서는 약간 증진되었다. 정제된 cellulase는 CMC, konjac, barley $\beta$-glucan과 lichenan을 가수분해하였으나 xylan, locust bean gum 및 p-nitrophenyl-$\beta$-glucopyranoside를 분해하지 못하였다. Cellooligosaccharides를 정제된 WL-12 cellulase로 분해하였을 때 cellobiose와 cellotriose가 주된 최종 반응산물로 관찰되었으며 cellobiose보다는 중합도가 큰 cellotriose, cellotetrasoe와 cellopentaose는 분해하였으나 cellobiose는 분해하지 못하는 것으로 확인되었다.

• 한국작물학회지
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• 제50권1호
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• pp.32-38
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• 2005

A long-term study initiated in 1989 at San-born Field, Columbia, Missouri, was designed to evaluate the affect of environmental factors, nitrogen application, and crop rotation on soybean (Glycine max [L.] Merr.) seed composition. Soybeans were grown as part of a four- year rotation which included corn (Zea maize L.), wheat (Triticum aestivum L.), and red clover (Trifolium pratense L.). Results from soil tests made prior to initiation of the study and subsequently every five years, were used to calculate application rates of nitrogen, phosphorus, and potassium necessary for target yield of pursuant crops. In the experimental design, nitrogen was applied to one-half of the plot on which the non-leguminous crop, either corn or wheat was grown. Analysis of soybean seed by near infrared reflectance spectroscopy collected over an 11-year period revealed a linear increase in protein and decrease in oil content. Application of nitrogen fertilizer to non-leguminous crops did not have an apparent effect on total protein or oil content of subsequent soybean crop. Analysis of soybean seed proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis in conjunction with computer­assisted densitometry revealed subtle changes in the accumulation of seed proteins. Immunoblot analysis using antibodies raised against the $\beta-subunit$ of $\beta-conglycinin$ showed a gradual increase in the accumulation of the 7S components during successive years of the experiment. A linear increase in temperature and decrease in rainfall was observed from the onset of data· collection. Higher temperatures during the growing season have been linked to increased protein and diminished oil content of soybean, thus changes observed in this study are possibly related to climatic conditions. However, crop rotation and subsequent changes in soil ecology may contribute to these observed changes in the seed composition.

• Restorative Dentistry and Endodontics
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• 제24권3호
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• pp.437-446
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• 1999

Porphyromonas endodontalis(P. endodontalis) is one of the important causative bacteria of pulpal and periapical disease. P. endodontalis has lipopolysaccharide(LPS) and it plays a major role in stimulating the synthesis and release of cytokines from immune cells and prostaglandin $E_2$ from host cells. The purpose of this study is to prepare LPS from P. endodontalis and to evaluate the effect of LPS on membrane permeability of fibroblast. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted. LPS was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Human periodontal ligament cell, colon fibroblast(CCD-18Co, KCLB 21459) and skin fibroblast(Detroit 551, KCLB 10110) were perfused with 0.01% P. endodontalis LPS solution, high concentration of $K^+$ solution and $Ca^{2+}$-free solution, $Ca^{2+}$ concentration ratio was measured by microfluorometry. 1. Intracellular $Ca^{2+}$ concentration was not changed in human periodontal fibroblast and skin fibroblast(Detroit 551) stimulated by P. endodontalis LPS. 2. Intracellular $Ca^{2+}$ concentration was increased in colon fibroblast(CCD-18Co) stimulated by P. endodontalis LPS. 3. Colon fibroblast(CCD-18Co) has voltage dependent $Ca^{2+}$ channel activated by high concentration of $K^+$ solution. 4. P. endodontalis LPS has no effect on the increase of intracellular $Ca^{2+}$ concentration during perfusion of $Ca^{2+}$-free solution.

• Restorative Dentistry and Endodontics
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• 제22권2호
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• pp.693-701
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• 1997

In 1992, Prevotella intermedia was shown to be comprised of another spoecies now known as Prevotella nigrescens. Strain ATCC 33563 is now designated the type strain of P. nigrescens while strain ATCC 25611 is remains the type strain of P. intermedia. The purpose of this study was to find the differences in protein profiles of P. intermedia and P. nigrescens, using sodium dodecyl sulfate polyacrylamide gel electrophoresis, which can be used for differentiation of those two species. A partial amino acid sequence of the 18.6 kDa protein band, which was specific in P. nigrescens, was also determined. The cellular proteins were extracted from the cell pellets of pure cultures of P. intermedia. and P. nigrescens by either sonication or being shaken continuously for 20 min at $21^{\circ}C$ with 1 % SDS or being boiled for 3 min with 1 % SDS. SDS-PAGE was performed according to the method of laemmli using either 12% (w/v) gels or 18% (w/v) gels. Results were as follows ; 1. The similar electrophoretic protein profiles were shown by 3 cellular protein extraction methods for each strain. (Fig. 1 and 2) 2. the 18.6 kDa band which was specific only in P. nigrescens could be used for the differentiation of P. intermedia. and P. nigrescens. (Fig. 1 and 2, Table 1) 3. A total of 4 different tryptic fragments from the 18.6 kDa protein were sequenced. the resulting amino acid sequences were fragment 1.GNPVNIGGEW, 2.FNVVR, 3.NYLT-VAPY, and 4.GGDNVTTYQVLPEIGYN. By comparison to the sequences of known proteins in the Swiss-Prot database and PIR database. 90 % matching between fragment 1 and serine hydroxymethyl transferase(P24060) in the Swiss-Prot, and 90% matching between fragment 1 and glycine hydroxymethyl transferase(S15203) in the PIR were shown, but the identity and function of the 18.6 kDa protein remains unknown.