• Anatomy and Cell Biology
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• 제56권4호
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• pp.552-561
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• 2023

The endocrinology of type 2 diabetes (T2D) and its predisposing factors have been studied extensively while its skeletal effects have received negligible research despite this being a global disease. The cellular and molecular association between proximal humeral fractures and T2D has not been fully elucidated. We aimed to study bone cell quantities and immunolabel osteogenic and antiosteogenic cytokines. The study used 12-week-old rats (23 males) consisting of 8 Sprague Dawley (SD) and 15 Zucker Diabetic Sprague Dawley (ZDSD). Weekly mass measurements were taken while fasting blood glucose levels were recorded every 2 weeks with oral glucose tolerance tests conducted once every 4 weeks. Upon termination at the age of 28 weeks, humeri were fixed in 10% buffered formalin, prior to decalcification in ethylenediaminetetraacetic acid. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 ㎛ thickness in a series for Haematoxylin and Eosin stain, and immunohistochemistry was performed with the anti-tartrate-resistant acid phosphatase (TRAP), anti-alkaline phosphatase (ALP), anti-bone morphogenetic protein 3 (BMP3), anti-transforming growth factor beta 1 (TGFβ1), anti-aged glycation end product (AGE) antibodies in the sequence. ZDSD rats had more adipocytes, BMP3 and AGEs expression with higher numbers of TRAP positive osteocytes and fewer ALP cells although no differences were found in TGFβ1 immunopositivity. We also found that T2D increases the number of AGEs immuno-positive cells, as well as its extracellular expression, thus providing a conducive environment for the interaction of the osteogenic cytokine and its antagonist to suppress osteoblastogenesis. ZDSD groups had higher adipocyte numbers therefore increased marrow adiposity in T2D.

• 한국미생물·생명공학회지
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• 제20권3호
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• pp.295-301
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• 1992

토양으로부터 alkaline protease 생성능이 강한 Streptomyces griseus HC-1141을 분리하였으며, 정제 효소의 최적작용 pH 와 온도는 8.0, 60'C 였으며, pH 7.0-9.0의 범위와 $60^{\circ}C$이하에서 안정하였다. 금속 이온중 $Mn^{2+}$, $Ca^{2+}$, 등에 의해 활성이 증대되었으나 $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Ba^{2+}$, $Fe^{2+}$ 등에 의해 효소 활성이 저해되었고, 효소활성 저해제 중 Epsilon-aminocaproic acid, 2,4-dinitrophenol, iodine 등에 의해서는 현전한 효소활성저해가 관찰되지 않았으나 ethylenediaminetetraacetic acid와 p-chloromercuribenzoic acid에 의해 활성저해가 관찰되어 효소분자 중 SH기가 활성에 어느정도 관여하는 metallo enzyme으로 추정되었다. 정제효소의 $K_m$, $V_{max}$ 및 활성화에너지는 $2.229{\times}10^{-4}$M, $46.08 {\mu}$g/min, 3.643 kcal/mol 이었으며 hemoglobin과 egg albumin보다 casein을 더 잘 가수분해하였다. 또한 여러 가지 detergent에 대하여 강한 저항성을 가지고 있었다.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.

• 분석과학
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• 제29권5호
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• pp.234-241
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• 2016

비글견 혈장 중 arsenite (As(III)), arsenate (As(V)), dimethylarsinic acid (DMA)와 monomethylarsonic acid (MMA)를 정량하기 위한 분석법의 유효성을 검증하기 위하여 본 연구를 수행하였다. 비소를 종 분리하기 위하여 액체크로마토그래피 (HPLC) 와 결합된 유도결합 플라즈마 질량분석기 (ICP/MS) 를 사용하였으며, 비소를 정량하는 검출기에서 스펙트럼 간섭을 최소화하기 위하여 산소(O₂)를 반응기체로 하는 DRC (dynamic reaction cell)모드를 이용하였다. 분석법의 유효성을 검증하는 항목으로 선택성, 직선성, 정확성, 정밀성, 생체시료효과, 회수율, 시스템 적합성, 희석의 타당성과 안정성 실험을 실시하였다. 선택성의 결과 정량에 방해되는 피크는 없는 것으로 확인 되었으며, 정량범위에서 평균 상관계수가 0.999 이상의 좋은 직선성을 보였다. 최저정량한계는 As(III), As(V)와 DMA는 5 ng/mL이였고, MMA는 20 ng/mL이였다. 생체시료의 영향, 시스템 적합성 과 회수율 항목을 통해 정량성에 대한 영향이 없음을 확인하여 비글견 혈장 중 비소 종을 분리하여 정량하는 분석법의 유효성을 검증하였다. 따라서 본 연구에서 제시한 분석법은 건강에 유해한 비소의 농도를 정량 및 평가하는데 적용될 것이다.

• 생약학회지
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• 제47권2호
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• pp.122-127
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• 2016

Methicillin-Resistant Staphylococcus aureus(MRSA) is a multidrug-resistant(MDR) strain. (+)-Usnic acid(UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against 17 different strains of the bacterium. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid(EDTA) was used. In the other hand, Sodium azide($NaN_3$) was used as inhibitors of ATPase. Against the 17 strains, the minimum inhibitory concentrations(MICs) of UA were in the range of $7.81-31.25{\mu}g/ml$. EDTA or $NaN_3$ cooperation against MRSA showed synergistic activity on cell wall. UA and in combination with EDTA and $NaN_3$ could lead to the development of new combination antibiotics against MRSA infection.

• Biomolecules & Therapeutics
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• 제10권2호
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• pp.104-113
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• 2002

The effects of enzyme inhibitors and penetration enhancers on the permeation of leucine enkephalin (Leu-Enk) and its synthetic analog, [${D-ala}^2$]-leucine enkephalinamide (YAGFL) across the nasal, rectal and vaginal mucosae were evaluated. Enzyme inhibitors and penetration enhancers employed for Leu-Enk permeation study were amastatin(AM), thimerosal(TM) and ethylenediaminetetraacetic acid disodium salt(EDTA), and sodium taurodihydrofusidate (STDHF). Those for YAGFL permeation study were TM, benzalkonium chloride(BC) and EDTA, and STDHF, sodium deoxycholate(SDC), sodium glycholate(SGC), glycyrrhizic acid ammonium salt (GAA), L-$\alpha$-Iysophosphatidylcholine(LPC) and mixed micelle (MM, STDHF: linoleic acid = 15 mM : 5 mM). The addition of TM alone on the donor and receptor solutions for Leu-Enk permeation study across all the three kinds of mucosae failed to inhibit the degradation; it completely degraded in 6 hrs, and no permeation occurred. However, with addition of three kinds of inhibitors together, the fluxes across nasal, rectal and vaginal mucosae were $\20.7{pm}2.5$>/TEX>,$\0.3{pm}0.05$>/TEX> and $\1.4{pm}0.5$ $\mu$\mid$textrm{m}$/$\textrm{cm}^2$/hr, respectively. Moreover, the addition of STDHF in the presence of the above three inhibitors enhanced permeation across nasal, rectal and vaginal mucosae 1.3, 15 and 1.3 times, respectively. YhGFL also degraded in the donor and receptor solutions rapidly as time went. With mixed inhibitors of TM and EDTA, the percents of YAGFL remaining in the donor solutions facing nasal, rectal and vaginal mucosae were 69.7, 69.8 and 79.8%, respectively; the percent permeated increased to 10, 2.1 and 5.7%, respectively. The addition of STDHF in the presence of either BC/EDTA or TW/EDTA increased the permeation 2.2, 11.0 and 2.9 times, and 2.21, 14.0 and 2.7 times for nasal, rectal and vaginal mucosae, respectively. With SDC, SGC, GAA, LPC ud MM in the presence of TM/EDTA increased permeation; especially, they increased permeation across vaginal mucosae effectively, and the enhancement factors were 12.5, 7.6, 8.7, 5.7 and 5.5, respectively. The degradation extent of YAGFL was correlated with protein concentrations in the epidermal and serosal extracts. The flux of YAGFL across nasal mucosa increased dose-dependently.

• 한국미생물·생명공학회지
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• 제44권4호
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• pp.470-478
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• 2016

지렁이의 장내로부터 분리한 미생물 중에서 키틴 가수분해 활성이 우수한 미생물을 선발하였으며, 이를 동정하여 Bacillus licheniformis GA9으로 명명하였다. B. licheniformis GA9이 생산하는 키틴 분해효소의 정제는 배양상등액 40-60% 황산암모늄 침전, 음이온교환 크로마토그래피, 겔 크로마토그래피를 사용하여 정제하였다. 최종적으로 정제한 키틴 분해효소는 45.2배로 정제되었고 효소단백질 회수율은 20.0%를 나타내었다. 정제한 키틴 분해효소의 분자량은 약 52.1 kDa으로 나타났으며, N-terminal amino acid sequencing 분석결과, 아미노산 서열은 D-S-G-K-N-G-K-I-I-R-Y-Y-P-IR로 확인되었다. 키틴 분해효소의 최적반응 pH와 pH 안정성을 측정한 결과, pH 5.0에서 최대 활성을 나타내었으며 pH 5.0-6.0에서 안정성을 나타내었다. 키틴 분해효소의 최적반응 온도와 온도안정성의 경우, $40^{\circ}C$에서 최대 활성을 나타내었으며, $60^{\circ}C$까지 60%의 잔존 활성을 나타내었다. 정제한 키틴 분해효소는 10 mM $Co^{2+}$ 금속이온에 의해 효소활성이 증가하였으며, $Fe^{2+}$과 $Cu^{2+}$ 금속이온에 의해 효소활성이 감소하였으나, EDTA 첨가시 감소한 효소활성이 일부 회복되었다. 정제한 효소의 $K_m$ 및 $V_{max}$는 각각 4.02 mg/ml와 0.52 mg/min이었다. 또한 키틴 분해효소는 생명공학, 생물의약, 농업, 식품영양 등 다양한 산업분야에서 응용이 가능하다.

• 한국지하수토양환경학회지:지하수토양환경
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• 제20권7호
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• pp.103-111
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• 2015

Soil washing with ethylenediaminetetraacetic acid (EDTA) is highly effective in the remediation of soils contaminated with heavy metals. The EDTA recycling process is a requisite for reducing the operating cost. The applicability of Na₂S addition on the precipitation of heavy metals from the spent soil washing solution and thereby recycling of EDTA was investigated. Addition of Na₂S into the single metal-EDTA and the mixed metal-EDTA solutions ([Na₂S]/[metal-EDTA] ratio = 30, reaction time = 30 min and pH = 7~9) was highly effective in the separation of Cu and Pb from metal-EDTA complexes, but not for Ni. The Zn removal efficiency varied with pH and slightly increased upto 40% as the reaction time increased from 0 to 240 min which was longer than those for Cu and Pb. Ca(OH)₂ was subsequently added to induce further precipitation of Zn and Ni and to reduce the Na₂S dose. At the [Na₂S]/[metal-EDTA] ratio of 10, the removal efficiencies of all heavy metals excluding Ni were above 98% with the dose of Ca(OH)₂ at 0.002, 0.006 and 0.008 g into 100 mL of Cu-, Pb- and Zn-EDTA solutions, respectively. However, Ca(OH)₂ addition was not effective for Ni-EDTA solution. A further research is needed to improve metal removal efficiency and subsequent EDTA recycling for the real application in field-contaminated soils.

• Clinical and Experimental Reproductive Medicine
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• 제41권2호
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• pp.92-95
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• 2014

Objective: This study sought to evaluate platelet volume indices (mean platelet volume [MPV], platelet distribution width [PDW], and platelet large cell ratio [P-LCR]) in varicocele patients, and compare it with platelet volume parameters in healthy controls. Methods: This cross-sectional study involved 2 groups: group 1 included 51 varicocele subjects and group 2 consisted of 50 healthy control subjects of similar ages. Peripheral venous blood samples were collected with ethylenediaminetetraacetic acid-K2 anticoagulant between 8:30 AM and 10 AM following an overnight fast. Platelet volume parameters (MPV, PDW, and P-LCR) were measured in both groups within 2 hours of sampling. Results: The mean PDW, MPV, and P-LCR were $13.9{\pm}2.5%$, $10.1{\pm}1.3fL$, and $27.3{\pm}7.8%$ in varicocele patients, respectively, and were $12.6{\pm}2.4%$, $9.3{\pm}1.1fL$, and $21.9{\pm}6.4%$ in the control group, respectively. The mean PDW, MPV, and P-LCR were significantly higher in the varicocele group than the control group. Conclusion: The results of the present study suggest that vascular components may play an important role in the pathophysiology of varicocele; therefore, there is a great need for prospective studies to confirm this relationship.

• 한국식품과학회지
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• 제31권4호
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• pp.1109-1114
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• 1999

식품에서 산화 방지제로 사용되고 있는 EDTA를 이용하여 Saccharomyces cerevisiae에 대한 항진균 활성을 조사하였다. 그 결과 일반적인 항진균 측정 조건에서 EDTA의 MIC(정균 농도) 및 MFC(살균 농도)값은 각각 200 ${\mu}g/mL$과 6,400 ${\mu}g/mL$로 나타났다. 또한 EDTA의 항진균 활성은 항세균 활성에서 알려진 바와 같이 높은 종균 농도, 산성 배지, 배지에 첨가된 금속 이온($Ca^{++},\;Mg^{++}$)에 의해 크게 감소하였다. 한편 EDTA를 천연 식품 양념으로부터 분리된 polygodial과 병용한 결과 Saccharomyces cerevisiae에 대한 그들의 항진균 활성은 상호 상승적으로 증가하였다. 또한 종균 농도, 배지 pH, 금속 이온($Ca^{++},\;Mg^{++}$)의 배지 첨가와 같은 측정 조건에 따른 EDTA의 항진균 활성 감소는 polygodial과 병용에 의해 크게 개선되었다.