• Title/Summary/Keyword: enzyme resource

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ADA Levels in Body Fluids as the Preferred Test to Rule Out Tuberculosis in Limited-resource Settings: Data from a Tertiary Care Hospital in Northern India

  • Rasool, Rafia;Rashid, Gowhar;Mir, Shafat Ahmad;Rather, Tahseen Bilal;Mudassar, Syed
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.3
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    • pp.167-172
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    • 2022
  • In clinical practice, the diagnosis of tuberculosis (TB) continues to be a challenge. The goal of this study was to evaluate the reliability and impact of adenosine deaminase (ADA) enzyme testing as a biochemical marker in the continued management of suspected tuberculosis in a limited resource setting hospital. The retrospective data were collected from 2018 to 2021 and comprised the results of all ADA test assays done in the laboratory. All types of body fluids received for ADA testing were analyzed. Over the course of two years, 1461 samples for ADA assay testing were received. The average age of the study population was 56.69±11.7 years, with males accounting for the majority of the subjects (55.72%). Pleural fluid (N=817, 55.92%) was the most common type of sample received for the ADA assay. 114 (13.95%) of the 817 pleural fluid samples were found to be positive. A survey was conducted to obtain physician's response regarding reliability on ADA testing. 100% of them reported the supportive role of ADA levels in the workup of patients with suspected tuberculosis. In a limited resource setting, the ADA test, in conjunction with clinical and other laboratory findings, can help physicians to initiate early treatment in hospitals for the benefit of patients.

Feeding broiler chicks with Schizosaccharomyces pombe-expressed phytase-containing diet improves growth performance, phosphorus digestibility, toe ash, and footpad lesions

  • Dang, De Xin;Chun, Seong Guk;Kim, In Ho
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1390-1399
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    • 2022
  • Objective: The objective of this study was to evaluate the effects of dietary supplementation of Schizosaccharomyces pombe (S. pombe) -expressed phytase on growth performance, apparent ileal digestibility, organ indexes, meat quality, toe ash, and footpad lesions score in broiler chicks. Methods: A total of 390 one-day-old broiler chicks were randomly assigned to 5 groups based on the initial body weight (42.15±0.17 g), there were 6 replicate cages per treatment and 13 birds (mixed sex) per cage. The experimental period was 45 days, including 4 periods (starter, days 1 to 10; grower, days 11 to 24; finisher 1, days 25 to 38; finisher 2, days 39 to 45). Dietary treatments were based on a corn-soybean meal-basal diet and supplemented with 500, 750, 1,000, and 1,500 FTU/kg S. pombe-expressed phytase. One phytase unit (FTU) was defined as the amount of enzyme that catalyzes the release of one micromole phosphate from phytate per minute at 37℃ and pH 5.5. Results: The inclusion of increasing levels of phytase in the diet linearly increased the body weight gain during days 1 to 10 (p = 0.001), 25 to 38 (p = 0.016), 39 to 45 (p = 0.018), and 1 to 45 (p = 0.004), feed intake during days 25 to 38 (p = 0.032), feed conversion ratio during days 1 to 10 (p = 0.001), 39 to 45 (p = 0.038), and 1 to 45 (p = 0.012), carcass weight (p = 0.035), toe ash (p<0.001), and apparent ileal phosphorus digestibility (p = 0.049). However, the footpad lesions score (p = 0.040) decreased linearly with the increase in phytase levels in the diet. Conclusion: Dietary supplementation of S. pombe-expressed phytase was beneficial to the growth performance, toe ash, apparent ileal phosphorus digestibility, and footpad lesions of broiler chicks in a dose-dependent manner.

Processing Condition of Seasoning Material of the Mixture of Laminaria and Enzyme-Treated Mackerel Meat (다시마와 효소처리 고등어육을 이용한 조미소재의 제조 조건)

  • Lee, Kang-Ho;Song, Byung-Kwon;Jeong, In-Hak;Hong, Byeong-Il;Jung, Byung-Chun;Lee, Dong-Ho
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.77-81
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    • 1997
  • In order to develop a new type of natural seasoning material combining fish meat with seaweed, a processing method of the mixture of enzyme treated mackerel meat and Laminaria powder was studied. Mackerel meat previously boiled and deboned was treated with proteolytic enzyme to enhance taste of meat by proper hydrolysis. The enzyme-treated meat was dried at $100{\pm}2^{\circ}C$ for 4 hrs, and finally mixed with kelp power, moistened in advance, plus binding agents (0.02% calcium carbonate) to aid the formation of pellets by extrusion. Boiled mackerel meat of enzyme treated (0.03% Protease-A) at $50^{\circ}C$ for 90 min was adequate to result an increase in 6 times of total free amino acid content and about 10% increase of taste-enhancing amino acids such as glutamic acid, glycine, arginine, lysine.

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Characteristics of Microbial Pretense far Application to Abolished Protein Resource (폐단백자원에 이용하기 위한 미생물 Protease의 특성)

  • Chun, Sung-Sook;Cho, Young-Je;Sung, Tae-Soo;Son, Jun-Ho;Choi, Cheong
    • Applied Biological Chemistry
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    • v.41 no.1
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    • pp.6-12
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    • 1998
  • To extract insoluble proteins and to improve functional properties of abolished proteins, a protease producing Aspergillus sp. MS-18 was isolated from soil. The enzyme was purified and its enzymological characteristics were investigated. It was found that production of protease reached to the maximum when the wheat brae medium containing, 3% arabinose, 0.5% polypepton, 0.1% $(NH_4)_2SO_4$ and 0.2% magnesium chloride was cultured for 3 days. Protease was purified 16.9 folds after ion exchange chromatography and gel filtration and the specific activity was 340.4 unit/mg. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. The molecular weight of protease was estimated to be 30,000. Crystalization form of purified protease was a stick shape with rounding edges. The optimum pH and temperature for the protease activity were 9.0 and $60^{\circ}C$, respectively. The enzyme was stable in pH 7.0-12.0 at $50^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$ and $Pb^{2+}$, whereas it was activited by $Na^+$, $Mg^{2+}$ and $Mn^{2+}$. The activity of the protease was inhibited by the treatment with ethylenediaminetetraacetic acid and phenylmethane sulfonyl fluoride. The result suggests that the purified enzyme is a serine protease with metal ion at active site. Km and Vmax of purified protease were $29.33\;{\mu}mole/L$ and $5.13\;{\mu}g/min$, respectively.

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Collagen-Induced Arthritis Analysis in Rhbdf2 Knockout Mouse

  • Lee, Min-Young;Kang, Ju-Seong;Go, Ryeo-Eun;Byun, Yong-Sub;Wi, Young Jin;Hwang, Kyung-A;Choi, Jae-Hoon;Kim, Hyoung-Chin;Choi, Kyung-Chul;Nam, Ki-Hoan
    • Biomolecules & Therapeutics
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    • v.26 no.3
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    • pp.298-305
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    • 2018
  • Rhomboid family member 2 gene (Rhbdf2) is an inactive homologue lacking essential catalytic residues of rhomboid intramembrane serine proteases. The protein is necessary for maturation of tumor necrosis factor-alpha ($TNF-{\alpha}$) converting enzyme, which is the molecule responsible for the release of $TNF-{\alpha}$. In this study, Rhbdf2 knockout (KO) mice were produced by CRISPR/CAS9. To see the effects of the failure of $TNF-{\alpha}$ release induced by Rhbdf2 gene KO, collagen-induced arthritis (CIA), which is the representative $TNF-{\alpha}$ related disease, was induced in the Rhbdf2 mutant mouse using chicken collagen type II. The severity of the CIA was measured by traditional clinical scores and histopathological analysis of hind limb joints. A rota-rod test and grip strength test were employed to evaluate the severity of CIA based on losses of physical functions. The results indicated that Rhbdf2 mutant mice showed clear alleviation of the clinical severity of CIA as demonstrated by the significantly lower severity indexes. Moreover, a grip strength test was shown to be useful for the evaluation of physical functional losses by CIA. Overall, the results showed that the Rhbdf2 gene has a significant effect on the induction of CIA, which is related to $TNF-{\alpha}$.

Effect of Polysaccharides from Astragalus membranaceus on Exercise-Induced Fatigue and Oxidative Damage in Skeletal Muscle in Exhaustive Exercise Animal Models (과도 운동에 의해 유발되는 피로 및 골격근 산화적 손상에 대한 황기 다당체의 효과)

  • Go, Eun Ji;Lee, Hannah;Park, Hyun Su;Kim, Soo Jin;Park, Yeong Chul;Seong, Eun Soo;Yu, Chang Yeon;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.271-280
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    • 2018
  • Background: Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.

Purification and Charactedrization of Cysteine Desulfhydrase from Streptomyces albidoflavus SMF301

  • Ryu, Jae-Gon;Kang, Sung-Gyun;Kim, In-Seop;Rho, Young-Taik;Lee, Sang-Hee;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.97-102
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    • 1997
  • Cysteine desulfhydrase (EC 4.4.1.1.) was purified from the culture supernatant of Streptomyces albidoflavus SMF301 by hydroxyapatite, gel filtration and Resource Q ion-exchange chromatography with a purification fold of six identical subunits. The enzyme was stabilized by dithiothreitol and pyridoxal 5'-phosphate during the purification procedures. The optimum pH and temperature were pH 8.6 and 35$^{\circ}C$, respectively. The N-terminal amino acid sequence was identified as A-P-L-P-T-A-D-V-R-S-D-P-G-Y-R-E-W-L-G-E-A-V. The purified cystein desulfhydrase had a high substrate specificity toward cysteine, and exhibited no cystahionine $\gamma$-lyase activity. The $K_m$ value for cysteine was determined to be 0.37 mM.

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Agaricus blazei Mycelial Liquid Culture Extract Containing Eritadenine Improves Benign Prostatic Hyperplasia-related Biochemical Markers in RWPE-1 Cells through Anti-inflammatory and Anti-oxidative Actions (RWPE-1 전립선세포에서 eritadenine을 함유한 신령버섯균사체 액체배양물의 항염증효과 및 항산화효과에 의한 전립선비대증 관련 biochemical marker 개선 효과)

  • Ha, Yeong Lae;Moon, Yun-Gu;Kim, Na-Hyun;Heo, Jeong Doo;Cho, Min Jung;Kim, Ye Ra;Kim, Young Suk;Kim, Jeong OK
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1147-1155
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    • 2018
  • Agaricus blazei mycelial liquid culture extract (ABMLCE) promoted the production of testosterone (TS) in TM-3 mouse Leydig testis cells. Now, we report that ABMLCE containing eritadenine (EA) as a minor constituent (15.3 mg/100 g) reduced $5{\alpha}-reductase$ 2 ($5{\alpha}-R2$) enzyme activity and dihydrotestosterone (DHT) content which are key constituents for the benign prostatic hyperplasia (BPH) inductions. RWPE-1 prostate cells were grown in a Keratinocyte serum-free medium (K-SFM) containing ABMLCE (0~50 ppm), EA (0~10 ppm,), and finasteride (FS $10{\mu}M$: a positive control) in a 24-well plate for 24 hr. Supernatants collected from cell-cultured media were used for the assay of $5{\alpha}-R2$, superoxide dismutase (SOD), catalase (CAT) and cyclooxygenase-2 (COX-2) enzyme activities, and for TS, DHT, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and $interleukin-1{\beta}$ ($IL-1{\beta}$) contents by their assay kits. The $5{\alpha}-R2$ activity and DHT content were proportionally reduced (p<0.05) to concentrations of ABMLCE. The SOD and CAT enzyme activities were significantly (p<0.05) elevated concomitant with ABMLCE concentrations, while COX-2, $TNF-{\alpha}$ and $IL-1{\beta}$ showed reverse results (p<0.05). Similarly, the effects of EA were similar to those of ABMLCE. Efficacies of ABMLCE 50 ppm and EA 10 ppm in $5{\alpha}-R2$ and DHT reduction were similar to those of $10{\mu}M$ FS. These results suggest that ABMLCE and EA reduced $5{\alpha}-R2$ and DHT through their anti-inflammatory and anti-oxidative actions. This implies that ABMLCE containing EA could be a beneficial material in the cure of BPH in humans.

Effect of varying ethanol concentrations on the extraction properties and physiological activity of Artemisia annua L. (에탄올 농도 조건이 개똥쑥 추출 및 생리활성에 미치는 영향)

  • Kim, Kyeoung Cheol;Kim, Ju-Sung
    • Korean Journal of Food Science and Technology
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    • v.52 no.2
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    • pp.130-137
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    • 2020
  • The present study was undertaken to compare the effects exerted by different extraction solvents on the extraction of active components, such as polyphenols and flavonoids, from the dried leaves of Artemisia annua L. Different extracts were prepared using a heating mantle. The extraction solvents used were distilled water, and 20, 40, 60, 80, and 99.5% ethanol solution. It was observed that the 40% ethanol solution yielded the most significant results in the extraction of various phytochemicals with phenol concentration of 154.8±0.28 mg of gallic acid equivalent/g and flavonoid content of 25.28±0.01 mg quercetin equivalent/g. However, based on the extraction solvent used, varying trends were observed in the antioxidant, enzyme inhibition, and bacterial inhibition analyses. It was concluded that the extraction solvent should be selected based on the purpose of use of the dried leaves of A. annua L.

Comparison of Extraction Methods for Determination of Vitamin K1 in Vegetables (채소류의 비타민 K1 분석을 위한 추출방법의 비교)

  • Kim, Hyeongi;Choi, Youngmin;Cho, Young-Sook;Sung, Jeehye;Ham, Hyeonmi;Lee, Junsoo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1791-1795
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    • 2014
  • The objective of this study was to compare two extraction methods for determination of vitamin K1 (phylloquinone) in vegetables. In addition, analytical method validation parameters such as accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and linearity were calculated to ensure the method's validity. Vitamin K1 was quantified by reversed-phase HPLC using post-column derivatization and fluorescence detection ($Ex{\lambda}=243nm$, $Ex{\lambda}=430nm$). Higher analytical values were observed using solvent extraction compared to those from the enzyme extraction method. The results from the method validation showed high linearity in the calibration curve with a coefficient of correlation ($R^2$) of 0.9994. The LOD and LOQ were 0.1335 and 0.2784 ng/injection volume ($50{\mu}L$), respectively. The inter-day precision and inter-day precision were 2.0% and 2.1%, respectively. Overall recovery was close to 100% (n=5). The phylloquinone contents ranged from 9.42 to $1,212.57{\mu}g/100g$. Our study provides reliable data on the phylloquinone contents in commonly consumed vegetables in Korea.