• Title/Summary/Keyword: Dialysis

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Purification and Characterization of an Invertase Produced with Saccharomyces cerevisiae JS59 Isolated from Home-made Wine (포도주에서 분리한 Saccharomyces cerevisiae JS59가 생성하는 Invertase의 정제 및 특성)

  • Yoo, Ji-Soo;Paik, Hyun-Dong;Kim, Soo-Young;Lee, Si-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1321-1327
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    • 2011
  • The microorganism producing an invertase (E.C. 3.2.1.26) was isolated from wine and tentatively identified as Saccharomyces cerevisiae by cellular fatty acid analysis. The invertase was purified to homogeneity by ammonium sulfate precipitant, dialysis, ion-exchange chromatography on DEAE-Sephadex A-50, and gel chromatography on Sephadex G-200 from the culture supernatant of Saccharomyces cerevisiae JS59. The specific activity and the purification fold of the purified invertase were 7620.9 unit/mg protein and 13.9, respectively. The molecular weight of the purified invertase was estimated to be 38.5 kDa by SDS-PAGE. The optimum pH and temperature for the invertase activity were pH 5 and $55^{\circ}C$, respectively. The invertase activity was relatively stable at pH 4~6 and temperature $55^{\circ}C$. The activity of invertase was inhibited by $Ag^{2+}$ and $Hg^{2+}$, but on the contrary, activated by $Co^{2+}$ and $Mn^{2+}$. Michaelis constant ($K_m$) for invertase reaction in sucrose solution was 11.5 mM. TLC analysis of the products produced in sucrose solution during invertase reaction showed the progressive presence of glucose and fructose in accordance with sucrose hydrolysis.

The need assessment of visiting health services in a rural community (농촌지역의 방문보건서비스에 대한 요구조사)

  • Lee, Kun-Sei;Kim, Chang-Yup;Kim, Yong-Ik;Shin, Young-Soo
    • Journal of agricultural medicine and community health
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    • v.19 no.1
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    • pp.41-52
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    • 1994
  • The aim of this study was to assess the people's need for visiting health services in a rural area. In recent years, the great concern for the visiting health services has aroused in Korea. Stratified cluster sample for a household survey was used to select 1,255(8.4%) Households from Yonchon county. This study was undertaken from July 26 to August 7 in 1993. Medically defined need, usually expressed by the prevalencies or rates of specific disease, was evaluated with the use of criteria established by medical and nursing professors and expressed by the percent of specific objects for the visiting health services. Perceived need represented by the acceptability for the visiting services and willingness to paying for it, also, evaluated. The major results were as follows : 1. Of the 348 patients with hypertension, 201 were the non-compliant patients, the rate of the non-compliant hypertension patients in 4,577 study population was 7.4%. 2. Of the 141 diabetic patients, 73 were the non-compliant patients, the rate of the non-compliant hypertension patients in 4,577 sample population was 2.7%. 3. The number of patients with severe musculo-skeletal disease was 24, the rate was 0.9% above the age 30. 4. Of the 514 elderly, 33 were the elderly without any family member, the rate was 8.4%. Those with severely decreased activity of daily living were 13. 5. Infants with high risks were 12, pregnant women and neonates were 5, patients discharged within 1 week and with special equipments such as peritoneal dialysis, stoma, TPN etc. were 17, and patients with cancer were 5. Total number of the objects needed visiting health services was 752(18.43%) of the 4,577 study population. Perceived need evaluated were as follows; The acceptance rate of visiting health services was 74.9%, The kinds of visiting health services such as family health protection and promotion, early detection of hypertension, physiotherapy, health education and counseling were needed in the order of high rate The price willing to pay for visiting health services per visit was about 3,000-5,000Won. In conclusion, Visiting health services programme to be developed should have priority to the prevention of complications of chronic disease such as hypertension, diabetes milletus, elderly disease and health promotion.

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Action Patterns of Chitinase and Separations of Chitooligosaccharides Produced by Chitinolytic Hydrolysis (키티나제에 의한 키토올리고당의 생성활성 규명과 올리고당의 당별 분리 생산)

  • Kim, Kwang
    • KSBB Journal
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    • v.17 no.1
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    • pp.100-105
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    • 2002
  • N-acetyl-D-glucosamine oligosaccharides [(GlcNAc)n] whose degree of polymer-ization is from one to ten (n=1-10) were fractionated by column chromatography on CM-Sephadex. Electro dialysis from a partially deacetylated chitosan hydrolysate prepared crudely with the N-acetyl-D-glucosaminidase(chitinase) and exo-N, N'-diacetylchito-biohydrolase(chitobiase) of Serratia marcescens QM B1466. Reducing sugar compositions and sequences of the N-acetyl-glucosamine oligosaccharides were identified by N-acetylation, randomly cleavage with chitinase and ego-splitting with chitobiase. N-acetyl-glucosamine heterochitooligosaccharides with glucosamine oligosaccharides, (GlcN)n at the reducing end residues together with $(GlcN)_1\sim(GlcN)_4$ were detected. Separation was accomplished by prefractionation with election by 0 to 1.0 M NaCl gradient solution. $(GlcNAc)_1 =4.25%,\; (GlcNAc)_2=4.49%,; (GlcNAc)_3=11.1%,\; (GlcNAc)_4=2.5%,$$ $(GlcNAc)_{5}$=0.64%, $(GlcNAc)_{6}$=2.12% and $(GlcNAc)_{7}$=1.21%, respectively, were crystallized after electrodialysis and lyophilization Each N-acetyl-D-glucosamine oligosaccharides content were detected by HPLC.

Phosphorylation as a Signal Transduction Pathway Related with N-channel Inactivation in Rat Sympathetic Neurons (N형 칼슘통로 비활성화와 연계된 세포 신호전달 체계로서의 인산화과정)

  • Lim Wonil;Goo Yong Sook
    • Progress in Medical Physics
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    • v.15 no.4
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    • pp.220-227
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    • 2004
  • In N-type $Ca^{2+}$ channels, the mechanism of inactivation - decline of inward current during a depolarizing voltage step- is still controversial between voltage-dependent inactivation and $Ca^{2+}$ -dependent inactivation. In the previous paper we demonstrated that fast component of inactivation of N-type calcium channels does not involve classic $Ca^{2+}$ -dependent mechanism and the slowly inactivating component could result from a $Ca^{2+}$ -dependent process. However, there should be signal transduction pathway which enhances inactivation no matter what the inactivation mechanism is. We have investigated the effect of phosphorylation on calcium channels of rat sympathetic neurons. Intracellular dialysis with the phosphatase inhibitors okadaic acid markedly enhanced the inactivation. The rapidly inactivating component is N-type calcium current, which is blocked by $\omega$-conotoxin GVIA. Staurosporine, a nonselective protein kinase inhibitor, prevented the action of okadaic acid, suggesting that protein phosphorylation is involved. More specifically lavendustin C, inhibitor of CaM kinase II, prevented the action of okadaic acid, suggesting that calmodulin dependent pathway is involved in inactivation process. It is not certain to this point whether phosphorylation process is inactivation itself. Molecular biological research regarding binding site should be followed to address the question of how the divalent cation binding site is related to phoshorylation process.

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Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Preparation and Characterization of Lithocholic Acid Conjugated Chitosan Oligosaccharide Nanoparticles for Hydrophobic Anticancer Agent Carriers (소수성 항암제의 전달체로 응용하기 위한 리소콜릭산이 결합된 키토산 나노입자의 제조와 특성)

  • Park, Jun-Kyu;Kim, Dong-Gon;Choi, Chang-Yong;Jeong, Young-Il;Kim, Myung-Yul;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Polymer(Korea)
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    • v.32 no.3
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    • pp.263-269
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    • 2008
  • To develop carriers of hydrophobic anticancer agents based on chitosan, chitosan oligosaccharide lactate (COS) was chemically modified with lithocholic acid (LA) which is one of the bile acids as a hydrophobic group. The physicochemical properties of the lithocholic acid conjugated chitosan nanoparticles (COS-LA) were investigated using $^1H$-NMR spectroscopy, dynamic light scattering (DLS) and spectrofluorophotometer. COS-LA-paclitaxel (CLs-Tx) nanoparticles loading paclitaxel as an anticancer agent were prepared by a dialysis method and its loading efficiency was measured through HPLC. On the basis of DLS results, the estimated particle sizes of CLs-Tx were around 300 nm. Also, the critical micelle concentration (CMC) was proven to be dependent on the degree of substitution of lithocholic acid. It showed that the CLs-Tx has the superior potential for the application as a paclitaxel carrier.

Effects of Addition of Chicory Extract on Starch Hydrolysis in vitro and Glucose Response in Healthy Subjects (치커리추출물 첨가가 in vitro 전분가수분해율 및 정상성인의 혈당반응에 미치는 영향)

  • Lee, Jung-Sun;Shin, Hyun-Kyung
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1295-1303
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    • 1997
  • This study determined the effects of addtition of chicory extract on the rate of starch hydrolysis in vitro and blood glucose response in healthy subjects. The rate of corn starch hydrolysis in the presence or absence of chicory extract was determined in an in vitro enzyme/dialysis system for 2hr. Additions of dried or roasted chicory extract (5%, w/w) to corn starch solution reduced the starch hydrolysis and significantly (p<0.05) decreased the area under hydrolysis curve by 16% and 18%, respectively. Groups of five to nine volunteers underwent 60 g glucose tolerance tests (GTT) with 2.5, 5, 10, 20% (w/w) dried or roasted chicory extracts. The addition of chicory extracts significantly (p<0.05) reduced blood glucose concentration during the GTT and reduced the mean peak rise and area under blood glucose curve. The glycemic indices of all dried chicory extract, $5{\sim}20%$ roasted chicory extract groups were significantly decreased compared with glucose control. Chicory extract is therefore likely to be useful in modifying postprandial hyperglycemia.

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The Relationship between Quality of Life and Depression, Anxiety in Hemodialysis Patients (혈액투석 환자의 삶의 질과 우울, 불안과의 관계)

  • Jeon, Yeong Ae
    • Journal of Internet of Things and Convergence
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    • v.4 no.1
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    • pp.7-22
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    • 2018
  • This descriptive investigation aims to identify a relationship between QOL, depression, and anxiety in hemodialysis patients. I conducted this research on 130 subjects aged 19 or above who have received dialysis for at least one month in the hemodialysis unit at N hospital in the city of S. However, it came to 112 as I took out those who either quit, died, or filled out a questionnaire in the incorrect manner. In regards to average points of QOL on a 100-point scale, it is 57.77, demonstrating quite low QOL. Concerning average points of depression and anxiety, it is 12.25 and 10.52, respectively, showing quite a high figure. With regard to a correlation, there is a significant correlation between the three factors in hemodialysis patients: there is a negative correlation between QOL and depression (r=-.782, p<.001); a negative correlation between QOL and anxiety (r=-.719, p<.001); and a positive correlation between depression and anxiety (r=.873, p<.001). The outcome of multiple regression analysis on factors affecting QOL shows that depression is the element influencing QOL (β=-.585). Therefore, this study is significant in providing basic data for establishing intervention strategies aiming to reduce levels of depression which has the strongest impact on QOL in the patients and for developing a nursing intervention program intending to improve their QOL.

Cloning, Expression, and Polymerization Assay of FtsZ Protein from Staphylococcus aureus (Staphylococcus aureus FtsZ의 클로닝, 발현 및 폴리머 형성 활성 분석)

  • Son, Sang Hyeon;Lee, Dong Yun;Kim, Ye Jun;Ko, Sooho;Cho, Seong Jun;Jung, Hyo Cheol;Lee, Hyung Ho
    • Microbiology and Biotechnology Letters
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    • v.40 no.3
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    • pp.274-277
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    • 2012
  • Cytokinesis is the final stage of cell division, dividing one mother cell into two daughter cells. For the cutting of a plasma membrane during bacterial cytokinesis, a tubulin homolog FtsZ protein is recruited from the cytoplasm to the division site. FtsZ protein polymerizes in a GTP-dependent manner and its N-terminal domain has a GTPase activity. In this study, we have begun to characterize FtsZ from Staphylococcus aureus (SA). Full-length SA FtsZ was cloned into pRSFDuet-1 vector and the clone was transformed into a BL21 (DE3) star cell. The recombinant SA FtsZ protein was purified using Ni-NTA affinity chromatography and dialysis. Using a spectrofluorometer, we showed that SA FtsZ undergoes a GTP-dependant polymerization in vitro. The polymer of the SA FtsZ protein disappeared after a few minutes, suggesting that the polymer is degraded as the GTP is consumed. This assay system may well be applied for inhibitor screening targeting S. aureus FtsZ.

Studies on the ${\beta}-Tyrosinase$ -Part 1. On the Enzymological Characteristics of ${\beta}-Tyrosinase$- (${\beta}-Tyrosinase$에 관한 연구 -제1보, ${\beta}-Tyrosinase$의 효소학적(酵素學的) 성질(性質)에 대하여-)

  • Kim, Chan-Jo;Nagasawa, Toru;Tani, Yoshiki;Yamada, Hideaki
    • Applied Biological Chemistry
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    • v.22 no.4
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    • pp.191-197
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    • 1979
  • ${\beta}-Tyrosinase$ was purified and crystallized from cells of Escherichia intermedia A-21 grown in a medium supplemented with 0.2% L-tyrosine. Molecular weight of its subunit, Km value and absorption spectra were determined. Crystallization methods were also studied to eliminate any unnecessary procedures. The results obtained were as follows: 1. The purification procedure included ammonium sulfate fractionation, dialysis against potassium phosphate buffer, pH 6.0 and pH 7.0, and DEAE-Sephadex column chromatography. In the column chromatography, 11 mg of protein was applied per ml of DEAE-Sephadex for efficiency. 2. Steps of protamine sulfate treatment and Sephadex G-150 gel filtration could be eliminated for this enzyme from the known procedures. 3. The purified enzyme was dissolved in 0.01M potassium phosphate buffer containing 2-mercaptoethanol, with a concentration of 20mg/ml. Crystalline enzyme, which appears as hexagonal rods, was obtained by adding solid fine powdered ammonium sulfate to the enzyme solution. 4. Absorption maxima of the enzyme appeared at 340 and 430nm when associated with pyridoxal phosphate. 5. Km value of the enzyme for L-tyrosine was $2.31{\times}10^{-4}M$ and the molecular weight of its subunit was determined by SDS-polyacrylamide electrophoresis to be approximately 50,000.

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