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Purification and Characterization of a Thrombolytic Enzyme Produced by a New Strain of Bacillus subtilis

  • Frias, Jorge;Toubarro, Duarte;Fraga, Alexandra;Botelho, Claudia;Teixeira, Jose;Pedrosa, Jorge;Simoes, Nelson
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.327-337
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    • 2021
  • Fibrinolytic enzymes with a direct mechanism of action and safer properties are currently requested for thrombolytic therapy. This paper reports on a new enzyme capable of degrading blood clots directly without impairing blood coagulation. This enzyme is also non-cytotoxic and constitutes an alternative to other thrombolytic enzymes known to cause undesired side effects. Twenty-four Bacillus isolates were screened for production of fibrinolytic enzymes using a fibrin agar plate. Based on produced activity, isolate S127e was selected and identified as B. subtilis using the 16S rDNA gene sequence. This strain is of biotechnological interest for producing high fibrinolytic yield and consequently has potential in the industrial field. The purified fibrinolytic enzyme has a molecular mass of 27.3 kDa, a predicted pI of 6.6, and a maximal affinity for Ala-Ala-Pro-Phe. This enzyme was almost completely inhibited by chymostatin with optimal activity at 48℃ and pH 7. Specific subtilisin features were found in the gene sequence, indicating that this enzyme belongs to the BPN group of the S8 subtilisin family and was assigned as AprE127. This subtilisin increased thromboplastin time by 3.7% (37.6 to 39 s) and prothrombin time by 3.2% (12.6 to 13 s), both within normal ranges. In a whole blood euglobulin assay, this enzyme did not impair coagulation but reduced lysis time significantly. Moreover, in an in vitro assay, AprE127 completely dissolved a thrombus of about 1 cc within 50 min and, in vivo, reduced a thrombus prompted in a rat tail by 11.4% in 24 h compared to non-treated animals.

Effects of Natural Herb Mixture on Anti-atopic Dermatitis and Skin Regeneration (천연 소재 복합물이 항아토피 피부염 및 피부재생에 미치는 영향)

  • Kim, Won-Sik;Sim, Boo-Yong;Kim, Dong-Hee
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.55-62
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    • 2017
  • Objectives : This study aims to evaluate the effects of natural herb mixutre (NHM) on atopic dermatitis and skin regeneration using in vivo test. Methods : NHM was prepared with DW. 25% of NHM was applied to skin lesion, where atopic dermatitis was induced by DNCB in NC/Nga mice. The levels of cytokines (IL-4, IL-5, IL-6, IL-13, TNF-a, and $IFN-{\gamma}$), and IgE in serum were measured by Luminex. Immune cells (WBC, eosinophil, lymphocyte, and monocyte) in blood were counted by coulter counter. The gross investigation of atopic dermatitis index score test were performed during the NHM treatment period. Also, the histopathological change of dorsal skin was observed by H&E and M&T staining. Results : NHM showed the levels of IL-4, IL-5, IL-6, IL-13, IgE, WBC, eosinophil, lymphocyte, and monocyte in serum or blood were significantly decreased. On the contrary, the productions of FGF, and VEGF were increased in the serum. Also, atopic dermatitis index score in NHM-treated mice were observed in the similar levels to those of normal group. Histological examination demonstrated that NHM suppressed immune cell infiltration and thickening of epidermis, meanwhile the extraction induced collagen production in the dorsal skin. Conclusion : This study demonstrated that NHM is appeared to be effective on atopic dermatitis and skin regeneration efficacy based on the observations with hematologic, gross, and histologic examinations. Therefore, we suggest that NHM could be effectively used as an external therapeutics against atopic dermatitis and a consequence skin damage.

Temperature and magnetic field dependent optical properties of superconducting $MgB_2$ thin film (초전도 $MgB_2$ 박막의 온도와 장기장의 변화에 따른 광학적 성질)

  • Jung, J. H.;Lee, H. J.;Kim, K. W.;Kim, M. W.;Noh, T. W.;Wang, Y. J.;Kang, W. N.;Jung, C. U.;Lee, Sung-Ik
    • Progress in Superconductivity
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    • v.3 no.1
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    • pp.31-35
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    • 2001
  • We investigated the temperature and magnetic field dependent optical properties of a$ MgB_2$ thin film in the far-infrared region. In the superconducting state, i.e. 5 K, we obtained the values of superconducting gap $2\Delta$ ~ 5.2 meV and $2\Delta$ $_{k}$ $B/T_{c}$ ~1.8. Although the value of$ 2\Delta$$B/T_{c}$ was nearly half of the BCS value, the $2\Delta$ seemed to follow the temperature dependence of the BCS formula. Under the magnetic field (H), the superconducting state became suppressed. Interestingly, we found that the normal state area fraction abruptly increased at low field but slowly increased at high field. It did not follow the H-dependences predicted for a s-wave superconductor (i.e. a linear dependence) nor for a s-wave one (i.e. $H^{1}$2/ dependence). We discussed the complex gap nature of $MgB_2$ in comparison with two gap and anisotropic s-wave scenarios.ios.

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Effect of ages and season temperatures on bi-surface shear behavior of HESUHPC-NSC composite

  • Yang Zhang;Yanping Zhu;Pengfei Ma;Shuilong He;Xudong Shao
    • Advances in concrete construction
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    • v.15 no.6
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    • pp.359-376
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    • 2023
  • Ultra-high-performance concrete (UHPC) has become an attractive cast-in-place repairing material for existing engineering structures. The present study aims to investigate age-dependent high-early-strength UHPC (HESUHPC) material properties (i.e., compressive strength, elastic modulus, flexural strength, and tensile strength) as well as interfacial shear properties of HESUHPC-normal strength concrete (NSC) composites cured at different season temperatures (i.e., summer, autumn, and winter). The typical temperatures were kept for at least seven days in different seasons from weather forecasting to guarantee an approximately consistent curing and testing condition (i.e., temperature and relative humidity) for specimens at different ages. The HESUHPC material properties are tested through standardized testing methods, and the interfacial bond performance is tested through a bi-surface shear testing method. The test results quantify the positive development of HESUHPC material properties at the early age, and the increasing amplitude decreases from summer to winter. Three-day mechanical properties in winter (with the lowest curing temperature) still gain more than 60% of the 28-day mechanical properties, and the impact of season temperatures becomes small at the later age. The HESUHPC shrinkage mainly occurs at the early age, and the final shrinkage value is not significant. The HESUHPC-NSC interface exhibits sound shear performance, the interface in most specimens does not fail, and most interfacial shear strengths are higher than the NSC-NSC composite. The HESUHPC-NSC composites at the shear failure do not exhibit a large relative slip and present a significant brittleness at the failure. The typical failures are characterized by thin-layer NSC debonding near the interface, and NSC pure shear failure. Two load-slip development patterns, and two types of main crack location are identified for the HESUHPC-NSC composites tested in different ages and seasons. In addition, shear capacity of the HESUHPC-NSC composite develops rapidly at the early age, and the increasing amplitude decreases as the season temperature decreases. This study will promote the HESUHPC application in practical engineering as a cast-in-place repairing material subjected to different natural environments.

The effect of typhoon translation speed and landfall angle on the maximum surge height along the coastline

  • Qian, Xiaojuan;Son, Sangyoung
    • Proceedings of the Korea Water Resources Association Conference
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    • 2021.06a
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    • pp.153-153
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    • 2021
  • Storm Storm event is one of major issues in South Korea due to devastating damage at its landfall. A series of statistical study on the historical typhoon records consistently insist that the typhoon translation speed (TS) is on slowdown trend annually, and thus provides an urgent topic in assessing the extreme storm surge under future climate change. Even though TS has been regarded as a principal contributor in storm surge dynamics, only a few studies have considered its impact on the storm surge. The landfall angle (LA), another key physical factor of storm surge also needs to be further investigated along with TS. This study aims to elucidate the interaction mechanism among TS, LA, coastal geometry, and storm surge synthetically by performing a series of simulations on the idealized geometries using Delft3D FM. In the simulation, various typhoons are set up according to different combinations of TS and LA, while their trajectories are assumed to be straight with the constant wind speed and the central pressure. Then, typhoons are subjected to make landfall over a set of idealized geometries that have different depth profiles and layouts (i.e., open coasts or bays). The simulation results show that: (i) For the open coasts, the maximum surge height (MSH) increases with increasing TS. (ii) For the constant bed level, a typhoon normal to the coastline resulted in peak MSH due to the lowest effect of the coastal wave. (iii) For the continental shelf with different widths, the slow-moving typhoon will generate the peak MSH around a small LA as the shelf width becomes narrow. (iv) For the bay, MSH enlarges with the ratio of L/E (the length of main-bay axis /gate size) dropping, while the greatest MSH is at L/E=1. These findings suggest that a fast-moving typhoon perpendicular to the coastline over a broad continental shelf will likely generate the extreme storm surge hazard in the future, as well as the slow-moving typhoon will make an acute landfall over a narrow continental shelf.

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HYPERSURFACES WITH PRESCRIBED MEAN CURVATURE IN MEASURE METRIC SPACE

  • Zhengmao Chen
    • Bulletin of the Korean Mathematical Society
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    • v.60 no.4
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    • pp.1085-1100
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    • 2023
  • For any given function f, we focus on the so-called prescribed mean curvature problem for the measure e-f(|x|2)dx provided thate-f(|x|2) ∈ L1(ℝn+1). More precisely, we prove that there exists a smooth hypersurface M whose metric is ds2 = dρ2 + ρ2d𝜉2 and whose mean curvature function is ${\frac{1}{n}}(\frac{u^p}{{\rho}^{\beta}})e^{f({\rho}^2)}{\psi}(\xi)$ for any given real constants p, β and functions f and ψ where u and ρ are the support function and radial function of M, respectively. Equivalently, we get the existence of a smooth solution to the following quasilinear equation on the unit sphere 𝕊n, $${\sum_{i,j}}({{\delta}_{ij}-{\frac{{\rho}_i{\rho}_j}{{\rho}^2+|{\nabla}{\rho}|^2}})(-{\rho}ji+{\frac{2}{{\rho}}}{\rho}j{\rho}i+{\rho}{\delta}_{ji})={\psi}{\frac{{\rho}^{2p+2-n-{\beta}}e^{f({\rho}^2)}}{({\rho}^2+|{\nabla}{\rho}|^2)^{\frac{p}{2}}}}$$ under some conditions. Our proof is based on the powerful method of continuity. In particular, if we take $f(t)={\frac{t}{2}}$, this may be prescribed mean curvature problem in Gauss measure space and it can be seen as an embedded result in Gauss measure space which will be needed in our forthcoming papers on the differential geometric analysis in Gauss measure space, such as Gauss-Bonnet-Chern theorem and its application on positive mass theorem and the Steiner-Weyl type formula, the Plateau problem and so on.

The Novel Label Free Staining Algorithm in Digital Pathology (차세대 디지털 병리를 위한 Label Free 디지털염색 알고리즘 비교 연구)

  • Seok-Min Hwang;Yeun-Woo Jung;Dong-Bum Kim;Seung Ah Lee;Nam Hoon Cho;Jong-Ha Lee
    • Journal of the Institute of Convergence Signal Processing
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    • v.24 no.1
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    • pp.76-81
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    • 2023
  • To distinguish cancer cells from normal cells, H&E (Hematoxylin & Eosin) staining is required. Pathological staining requires a lot of money and time. Recently, a digital dyeing method has been introduced to reduce such cost and time. In this paper, we propose a novel digital pathology algorithms. The first algorithm is the Pair method. This method learns the dyed phase image and unstained amplitude image taken by FPM (Fourier Ptychographic Microscopy) and converts it into a dyed amplitude image. The second algorithm is the unpair method. This method use the stained and unstained fluorescence microscopic images for modeling. In this study, digital staining was performed using a generative adversarial network (GAN). From the experimental results, we noticed that both the pair and unpair algorithms shows the excellent performance.

Hematological characteristics and endocrine profiles of cloned dromedary camels (Camelus dromedaries)

  • Young-Bum Son;Mohammad Shamim Hossein;Yeon Ik Jeong;Mina Kang;Huijeong Kim;Yura Bae;Kung Ik Hwang;Alex Tinson;Singh Rajesh;Al Shamsi Noura;Woo Suk Hwang
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.12-18
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    • 2024
  • Background: Somatic cell nuclear transfer (SCNT) is a prominent technology that can preserve superior genetic traits of animals and expand the population in a short time. Hematological characters and endocrine profiles are important elements that demonstrate the stability of the physiological state of cloned animals. To date, several studies regarding cloned camels with superior genes have been conducted. However, detailed hemato-physiological assessments to prove that cloned camels are physiologically normal are limited. In this study, We evaluated the hemato-physiological characteristics of cloned male and female dromedary camels (Camelus dromedaries). Methods: Therefore, we analyzed variations in hematological characteristics and endocrine profiles between cloned and non-cloned age-matched male and female dromedary camels (Camelus dromedaries). Two groups each of male and female cloned and non-cloned camels were monitored to investigate the differences in hemato-physiological characteristics. Results: All the animals were evaluated by performing complete blood count (CBC), serum chemistry, and endocrinological tests. We found no significant difference between the cloned and non-cloned camels. Furthermore, the blood chemistry and endocrine profiles in male and female camels before maturity were similar. Conclusions: These results suggest that cloned and non-cloned camels have similar hematological characteristics and endocrine parameters.

INHIBITORY EFFECT OF OCTYL-PHENOL AND BISPHENOL A ON CALCIUM SIGNALING IN CARDIOMYOCYTE DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS

  • J.-H. LEE;Y.-M. YOO;E.-M. JUNG;CH. AHN;E.-B. JEUNG
    • The Korean Journal of Physiology and Pharmacology
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    • v.70 no.3
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    • pp.435-442
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    • 2019
  • Endocrine-disrupting chemicals (EDCs) have structures similar to steroid hormones and can interfere with hormone synthesis and normal physiological functions of reproductive organs. For example, sex steroid hormones influence calcium signaling of the cardiac muscle in early embryo development. To confirm the effect of progesterone (P4), octyl-phenol (OP), and bisphenolA(BPA) on early differentiation of mouse embryonic stem cells(mESCs) into cardiomyocytes, mESCs were treated with P4, OP, and BPA two days after attachment and media were replaced every two days. In addition, cells were treated with mifepristone (RU486), a synthetic steroid that has an affinity for progesterone receptor (Pgr), for one day starting on day 11. Beating ratio was decreased with P4, OP, and BPA treatment. The Pgr mRNA level was significantly increased in the P4-, OP- and BPA-treated groups. However, the mRNA level of the calcium channel gene (Trpv2), contraction-related genes (Ryr2, Cam2, and Mylk3) and cardiac development and morphogenesis genes (Rbp4, Ly6e, and Gata4) were significantly decreased in the P4-, OP-, and BPA-treated groups. Interestingly, treatment with RU486 rescued the altered calcium channel gene, contraction-related genes, and cardiac development and morphogenesis genes. P4, OP, and BPA treatments reduced the intracellular calcium level. Taken together, these results indicate that EDCs (OP and BPA) has a structure similar to that of endogenous steroid hormones such as progesterone and estrogen, and OP and BPA act like progesterone to inhibit and disrupt cardiomyocyte differentiation of mESCs.

Geometric Analysis of Minor Faults and Paleostress Reconstruction around the Dongnae Fault (동래단층 주변 소단층의 분포 특성과 고응력장 복원)

  • 조용찬;장태우;이정모
    • Journal of the Earthquake Engineering Society of Korea
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    • v.2 no.4
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    • pp.41-52
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    • 1998
  • The Dongnae Fault in the southeastern part of the Korean Peninsular is not a single fault but a complex fault zone composed of numerous minor faults. In order to deduce the paleostress tensor evolving the Dongnae Fault, we measured 329 faults in outcrops around the fault zone and analyzed the geometries of them. Most of them are steeply dipping(>65˚) and fall into three groups striking N10E, N30E and N70E. More than one half of them show the rakes less than 30˚ Paleostress tensor analysis using the collected fault data has been conducted with the Angelier's direct inversion method and the Choi's method. As result, four different principal paleostress axes each of which subtends an independent tectonic event are found. They are; (1) NNE-SSW compression and ESE-WNW extension (Event I), (2) NNE-SSW extension (Event II), (3) ESE-WNW extension (Event III) and (4) ENE-WSW compression and NNW-SSE extension (Event IV) in chronology. Therefore, the tectonic movement around the Dongnae Fault was firstly governed by strike-slip faulting related to Event I. Afterward, normal faults were formed by Event II and Event III. Finally, the dextral strike-slip faults along the major trace of the Dongnae Fault were formed in NNE direction related to Event IV.

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