• Title/Summary/Keyword: Alkaline digestion

Search Result 58, Processing Time 0.027 seconds

IME EFFECT OF PULSED ELECTROMAGNETIC FIELD ON THE CULTURED CALVARIAL CELLS OF RAT (맥동 전자기장이 백서의 배양 두개관세포에 미치는 영향)

  • Choi, Byung-Taek;Yang, Won-Sik
    • The korean journal of orthodontics
    • /
    • v.20 no.3 s.32
    • /
    • pp.499-517
    • /
    • 1990
  • Electrical stimulation among several factors that influence bone remodeling has been studied by many investigators with great enthusiasm in orthodontic field. The action mechanisms of Pulsed Electromagnetic Field (PEMF) are different from those of the conventional electrode application method in that PEMF induces endogenous current in the living tissues. PEMF is known to have the healing effect in nonunion of bone and osteoporosis. It is widely used in orthopaedic scopes and the possibility of using the method in clinical orthodontics Is also conceivable. But the exact mechanisms by which the PEMF exerts its effects are not clearly understood. Therefore, the author wanted to see the effect of PEMF on five groups of rat calvarial cells obtained by sequential enzyme digestion method, and observed the changes in enzyme activation, collagen synthesis and $^3H-thymidine$ incorporation. The results were as follows: 1. Under the effect of PEMF, there were no changes in the alkaline phosphatase activity in five groups of cell populations. 2. Both the PEMF group and the PTH with PEMF group shelved no changes in acid phosphatase activities and there were no differences between two experimental groups. 3. Under the effect of PEMF, there was significant increase of collagen synthesis in the group V cell population. 4. Under the effect of PEMF, there were significant increases of $^3H-thymidine$ incorporation in the group IV and V cell populations.

  • PDF

DNA Shuffling of aprE Genes to Increase Fibrinolytic Activity and Thermostability

  • Yao, Zhuang;Jeon, Hye Sung;Yoo, Ji Yeon;Kang, Yun Ji;Kim, Min Jae;Kim, Tae Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • v.32 no.6
    • /
    • pp.800-807
    • /
    • 2022
  • Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.

The study on increasing of biodegradability by pre-treatment of municipal wasted sludge in anaerobic digestion process (도시하수슬러지의 전처리에 따른 혐기성 소화공정의 생분해율 향상에 관한 연구)

  • Kang, Chang-Min
    • Journal of the Korea Organic Resources Recycling Association
    • /
    • v.10 no.1
    • /
    • pp.87-95
    • /
    • 2002
  • The slow degradation rate of sludge in anaerobic digestion is due to rate-limiting step of sludge hydrolysis. To upgrading of sludge hydrolysis and biodegradabiliry, the pre-treatment had been carried out using acidlc (pH 1.5, 3, 4, 5) and alkaline (pH 9, 10, 13), thermal (50, 100, 150, $200^{\circ}C$), and ultrasonic treatment (400W, 20kHz, 15, 20, 25, 30, 40, 50, 60, 90min). In the best conditions of each treatment, the Soluble SCOD Ratio(%)of treated/untreated sample were increased 102% in acid (pH5), 986% in alkali (pH 13), 595% in thermal ($200^{\circ}C$) and 1123% in ultrasonic (35min) treatment. As the result, the ultrasonic treatment was most effective, followed by alkali, thermal, acid treatment. In the effects of total gas productivity in vial test, the thermal ($200^{\circ}C$) pre-treatment was the highest, followed by thermal ($150^{\circ}C$), ultrasonic (90min), alkaline (pH 9), and ultrasonic (50min). We compared untreated samples and the most efficient pre-treatment samples(at $200^{\circ}C$, for 30min) on gas productivity with changes of HRT in continuous experiments IN thermal treated samples were 2.5 times in SCOD, 2 times in soluble protein and 3.3 times high in soluble carbohydrate than untreated ones. In gas productivity, the thermal treated samples were average 2 times high than untreated ones. And HRT 7 days was most effective. followed by HRT 10, HRT 15 days. But The gas productiviry of HRT 2.5 days was less than untreated, the reason of low gas productivity was come from high organic acids accumulation within reactor.

  • PDF

Studies on Solvent Extraction and Analytical Application of Metal-dithizone Complexes(I). Separation and Determination of Trace Heavy Metals in Urine (Dithizone 금속착물의 용매추출 및 분석적 응용(제1보). 뇨중 흔적량 중금속 원소의 분리 정량)

  • Jeon, Moon-Kyo;Choi, Jong-Moon;Kim, Young-Sang
    • Analytical Science and Technology
    • /
    • v.9 no.4
    • /
    • pp.336-344
    • /
    • 1996
  • The extraction of trace cobalt, copper, nickel, cadmium, lead and zinc in urine samples of organic and alkali metal matrix into chloroform by the complex with a dithizone was studied for graphite furnace AAS determination. Various experimental conditions such as the pretreatment of urine, the pH of sample solution, and dithizone concentration in a solvent were optimized for the effective extraction, and some essential conditions were also studied for the back-extraction and digestion as well. All organic materials in 100 mL urine were destructed by the digestion with conc. $HNO_3$ 30 mL and 30% $H_2O_2$ 50 mL. Here, $H_2O_2$ was added dropwise with each 5.0 mL, serially. Analytes were extracted into 15.0 mL chloroform of 0.1% dithizone from the digested urine at pH 8.0 by shaking for 90 minutes. The pH was adjusted with a commercial buffer solution. Among analytes, cadmium, lead and zinc were back-extracted to 10.00 mL of 0.2 M $HNO_3$ from the solvent for the determination, and after the organic solvent was evaporated, others were dissolved with $HNO_3-H_2O_2$ and diluted to 10.00 mL with a deionized water. Synthetic digested urines were used to obtain optimum conditions and to plot calibration-eurves. Average recoveries of 77 to 109% for each element were obtained in sample solutions in which given amounts of analytes were added, and detection limits were Cd 0.09, Pb 0.59, Zn 0.18, Co 0.24, Cu 1.3 and Ni 1.7 ng/mL, respectively. It was concluded that this method could be applied for the determination of heavy elements in urine samples without any interferences of organic materials and major alkaline elements.

  • PDF

In Vitro Isolation and Proliferation of Mouse Male Germ-Line Stem Cells (생쥐 생식줄기세포의 체외 분리 및 증식)

  • 김수경;김계성
    • Journal of Embryo Transfer
    • /
    • v.18 no.3
    • /
    • pp.243-248
    • /
    • 2003
  • Sperrnatogenesis, the process by which the male germ-line stem cells(GSCs; type A spermatogonia) divide and differentiate to produce the mature spermatozoa, occurs in the seminiferous tubules of the testis. The GSCs proliferate actively to produce two types of cells: other GSCs and differentiating spermatogonia. GSCs have unipotentcy, devoted solely to the generation of sperm. The function of GSCs has broad implications for development, disease, and evolution. Spermatogenesis is fundamental for propagation of species and the defects of this system can result in infertility or disease. The ability to identify, isolate, culture, and alter GSCs will allow powerful new approaches in animal transgenesis and human gene therapy relating to infertility. Until recently, research on stem cells in the testis has been limited because of technical difficulties in isolating and identifying these cell populations. Here, we were trying to find out optimal conditions for in vitro culture of GSCs for identifying and isolating GSCs. We collected mouse GSCs from 3-days old mouse by two-step enzyme digestion method. GSCs were plated and grown on mouse embryonic fibroblasts in Dulbecco's modified Eagle's medium (DMEM) containing 15% fatal bovine serum, 10 mM 2-mercaptoethanol, 1% non-essential amino acids, 1 ng/$m\ell$ bFGF, 10 $\mu$M forskolin, 1500 U/$m\ell$ human recombinant leukemia inhibitory factor (LIF). Over a period 3∼5 days, GSCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells. After 5th passages, cells within the colonies continued to be alkaline phosphatase and Oct-4 positive and tested positive against a panel of two immunological markers(Integrin $\alpha$ 6 and Integrin $\beta$ 1) that have been recognized generally to characterize GSCs. SSEA-1, SSEA-3, and SSEA-4 also showed positive signals. Based on our data, these GSCs-derived cultures meet the criteria for GSCs itself and even other pluripotent stem cells. We reported here the establishment of in vitro cultures from mouse male GSCs.

Effect of tannins and cellulase on growth performance, nutrients digestibility, blood profiles, intestinal morphology and carcass characteristics in Hu sheep

  • Zhao, M.D.;Di, L.F.;Tang, Z.Y.;Jiang, W.;Li, C.Y.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.32 no.10
    • /
    • pp.1540-1547
    • /
    • 2019
  • Objective: This study was conducted to evaluate the effects of tannins and cellulase on growth performance, nutrient digestibility, blood profiles, intestinal morphology, and carcass characteristics in Hu sheep. Methods: A total of 48 three-month-old meat Hu sheep ($25.05{\pm}0.9kg$) were blocked based on body weight, and randomly allotted to 4 treatments with 3 replicates of 4 sheep each. The experiment lasted for 80 d, and dietary treatments were as follows: i) CON, control diet; ii) TAN, CON+0.1% tannins; iii) CEL, CON+0.1% cellulase; iv) TAN+CEL, CON+0.1% tannins and 0.1% cellulase. Results: Compared with CON, CEL, and TAN+CEL had greater (p<0.05) final body weight (FBW) and average daily gain but lower (p<0.05) feed conversion ratio, while FBW of TAN+CEL was lower (p<0.05) than that of CEL. The apparent total tract digestibility (ATTD) of dry matter in TAN, CEL, and TAN+CEL groups were higher (p<0.05) than that in CON. CEL and TAN+CEL groups had greater (p<0.05) ATTD of crude fiber compared with TAN and CON, while TAN group had lower (p<0.05) ATTD of crude protein than other treatments. TAN, CEL, and TAN+CEL groups increased (p<0.05) serum globulin and alkaline phosphatase but decreased (p<0.05) albumin/globulin. Serum total protein was greatest for TAN+CEL, intermediate for TAN and CEL and least for CON (p<0.05). TAN+CEL group increased (p<0.05) dressing percentage compared with CON, while the backfat thickness of CEL was lower (p<0.05) than that of CON. The villus height of jejunum and ileum in CEL and TAN+CEL groups were greater (p<0.05) than that in CON, and the crypt depth and villus height: crypt depth of jejunum were increased (p<0.05) in TAN, CEL, and TAN+CEL groups. Conclusion: The addition of tannins and cellulase together promoted nutrient digestion, liver protein synthesis and intestinal development and thus improved growth performance and carcass characteristics.

Antioxidant, Anti-thrombosis, and Lipid Accumulation Inhibition Activities of Different Dried Vinegars (3종 분말식초의 항산화, 항혈전 및 항비만 활성)

  • Jong-Kyu Lee;Jong-Sik Kim;Ho-Yong Sohn
    • Journal of Life Science
    • /
    • v.34 no.5
    • /
    • pp.296-303
    • /
    • 2024
  • Vinegar is a fermented food product created by fermenting various sugar- and starch-containing ingredients with microorganisms. It contains a variety of organic acids, sugars, amino acids, esters, and other compounds that contribute to its unique sensory properties. Vinegar is known for its potential benefits, including aiding digestion, lowering blood sugar levels, anti-obesity effects, and antioxidant properties. It is also believed to contribute to improving alkaline body conditions. This study was conducted to develop functional dried vinegar powder from naturally fermented vinegars. Unripe apple, brown rice, and black chokeberry (aronia) were fermented using Gluconacetobacter xylinus for 90-180 days. The filtrate vinegar was spray dried with 37.46% maltodextrin, 5% glucose, 1% citric acid, and 0.04% vitamin C. Analysis of the acidity, color difference, water and soluble solid content, and heat stability of dried vinegar (DV) confirmed that spray drying is a suitable method for powder production. Moreover, the DVs exhibited excellent sensory attributes and solubility. Among the DVs, aronia-DV showed the highest 1,1-diphenyl-2-picryl hydrazyl and 2,2-azobis (3- ethylbenzothiazoline-6-sulfonate) radical scavenging activity (36.7% and 75.3%) and reducing power (0.334) at 0.5 mg/ml concentration, respectively. The nitrite scavenging activity was highest in brown unripe apple-DV, followed by aronia-DV and brown rice-DV. In the anti-thrombosis activity assay, aronia-DV showed the highest prothrombin inhibition. The brown rice-DV exhibited lipid accumulation inhibitory activity in 3T3-L1 adipocytes without cell cytotoxicity. Our results suggest the potential for commercialization of dried vinegar, highlighting its diverse benefits and applications.

Pharmacological and Biochemical Characterization of Cells Isolated from Fetal Rat Calvaria (백서태자두개관에서 분리한 세포의 약리학적 및 생화학적 특성에 관한연구)

  • Han, Nam-Soo;Cheong, Dong-Kyun;Mori, Masakazu
    • The Korean Journal of Pharmacology
    • /
    • v.26 no.2
    • /
    • pp.193-207
    • /
    • 1990
  • Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

  • PDF