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Crystal Structure and Biochemical Analysis of a Cytochrome P450 Steroid Hydroxylase (BaCYP106A6) from Bacillus Species

  • Ki-Hwa Kim;Hackwon Do;Chang Woo Lee;Pradeep Subedi;Mieyoung Choi;Yewon Nam;Jun Hyuck Lee;Tae-Jin Oh
    • Journal of Microbiology and Biotechnology
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    • 제33권3호
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    • pp.387-397
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    • 2023
  • Cytochrome P450 (CYP) is a heme-containing enzyme that catalyzes hydroxylation reactions with various substrate molecules. Steroid hydroxylases are particularly useful for effectively introducing hydroxyl groups into a wide range of steroids in the pharmaceutical industry. This study reports a newly identified CYP steroid hydroxylase (BaCYP106A6) from the bacterium Bacillus sp. and characterizes it using an in vitro enzyme assay and structural investigation. Bioconversion assays indicated that BaCYP106A1 catalyzes the hydroxylation of progesterone and androstenedione, whereas no or low conversion was observed with 11β-hydroxysteroids such as cortisol, corticosterone, dexamethasone, and prednisolone. In addition, the crystal structure of BaCYP106A6 was determined at a resolution of 2.8 Å to investigate the configuration of the substrate-binding site and understand substrate preference. This structural characterization and comparison with other bacterial steroid hydroxylase CYPs allowed us to identify a unique Arg295 residue that may serve as the key residue for substrate specificity and regioselectivity in BaCYP106A6. This observation provides valuable background for further protein engineering to design commercially useful CYP steroid hydroxylases with different substrate specificities.

ISFRNet: A Deep Three-stage Identity and Structure Feature Refinement Network for Facial Image Inpainting

  • Yan Wang;Jitae Shin
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • 제17권3호
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    • pp.881-895
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    • 2023
  • Modern image inpainting techniques based on deep learning have achieved remarkable performance, and more and more people are working on repairing more complex and larger missing areas, although this is still challenging, especially for facial image inpainting. For a face image with a huge missing area, there are very few valid pixels available; however, people have an ability to imagine the complete picture in their mind according to their subjective will. It is important to simulate this capability while maintaining the identity features of the face as much as possible. To achieve this goal, we propose a three-stage network model, which we refer to as the identity and structure feature refinement network (ISFRNet). ISFRNet is based on 1) a pre-trained pSp-styleGAN model that generates an extremely realistic face image with rich structural features; 2) a shallow structured network with a small receptive field; and 3) a modified U-net with two encoders and a decoder, which has a large receptive field. We choose structural similarity index (SSIM), peak signal-to-noise ratio (PSNR), L1 Loss and learned perceptual image patch similarity (LPIPS) to evaluate our model. When the missing region is 20%-40%, the above four metric scores of our model are 28.12, 0.942, 0.015 and 0.090, respectively. When the lost area is between 40% and 60%, the metric scores are 23.31, 0.840, 0.053 and 0.177, respectively. Our inpainting network not only guarantees excellent face identity feature recovery but also exhibits state-of-the-art performance compared to other multi-stage refinement models.

Comparison of Ventral Midline and Right Flank Approaches of Ovariohysterectomy in Bitches

  • Ishwor Dhakal;Bharata Regmi;Bablu Thakur;Ishwari Tiwari;Shraddha Tiwari;Yeonsu Oh;Manoj K. Shah
    • 한국임상수의학회지
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    • 제40권1호
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    • pp.25-30
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    • 2023
  • The ventral midline approach (VMA) and right flank approach (RFA) are common procedures for the sterilization of bitches. This study compared the different parameters viz. total duration of surgery, recovery time, and length of the incision as well as body temperature, heart rate, respiration rate, and SpO2 in each approach. Twenty (20) bitches were divided randomly for the RFA and VMA. Meloxicam (0.2 mg/kg) was administered subcutaneously half an hour before the induction to provide preemptive analgesia. Diazepam and ketamine were administered intravenously at dose rates of 0.25 mg/kg and 2.5 mg/kg, and 0.17 mg/kg and 3.33 mg/kg, respectively to produce and maintain anesthesia. Each parameter was recorded at the pre-operative, operative and post-operative times. The average duration of surgery and length of incision of RFA (16.1 ± 5.13 min and 2.44 ± 0.83 cm) were significantly lower (p < 0.05) than the VMA (21.3 ± 5.48 min and 3.53 ± 0.7 cm). The operated bitches showed hypothermia (p < 0.05) at 1 hour compared to baseline and 24 hours of surgery. Heart and respiration rates increased significantly (p < 0.05) during traction and severing of ovarian ligaments in bitches within the RFA group, but there was no significant difference within VMA approaches. The sedation score was significantly higher (p < 0.05) at 1 hour after surgery in both approaches. Based on the duration of surgery and length of incision RFA approach was quick and minimal skin wound. Further studies on bitches considering molecular investigations of surgical stress are imperative.

서비스 사용자의 능동적 피싱 사이트 탐지를 위한 트레이스 백 기반 인증 모델 (A Traceback-Based Authentication Model for Active Phishing Site Detection for Service Users)

  • 백용진;김현주
    • 융합보안논문지
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    • 제23권1호
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    • pp.19-25
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    • 2023
  • 현재 네트워크 환경은 초기의 단방향 정보제공 서비스에서 실시간 양방향 서비스를 제공하고 있다. 이에 따라 웹 기반의 정보 공유 형태는 사용자 상호간 다양한 지식 제공과 서비스가 가능하다. 그렇지만 이러한 웹 기반의 실시간 정보 공유 환경은 네트워크 취약점을 악용한 불법적인 공격자들에 의해 그 피해 사례를 빠르게 증가시키고 있다. 특히 피싱 공격을 시도하는 공격자들의 경우 특정 웹 페이지 서비스가 필요한 사용자들에게 위/변조된 웹 페이지를 생성시킨 다음 해당 웹 페이지에 대한 링크를 유도한다. 본 논문은 사이트 위/변조 여부를 기존의 수동적인 서버 기반 탐지 방식이 아닌 사용자가 직접 능동적으로 특정 사이트에 대한 위/변조 여부를 분석할 수 있도록 하였다. 이를 위해 트레이스 백 정보를 이용하여 불법적인 웹 페이지 접속을 유도하는 공격자의 위장된 웹 페이지를 탐지하여 정상 사용자들의 중요한 개인 정보 유출을 방지할 수 있도록 하였다.

Investigation of Enzymatic Activities in Marine Algae-Derived Fungi

  • Dawoon Chung;Woon-Jong Yu;Hyeong Seok Jang;Yong-Min Kwon;Seung Seob Bae;Grace Choi
    • 한국해양생명과학회지
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    • 제8권1호
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    • pp.56-67
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    • 2023
  • Marine macroalgae are important in coastal ecosystems and interact with marine microorganisms. In this study, we isolated fungi from seven types of marine macroalgae including Cladophora sp., Gloiopeltis furcate, Gracilariopsis chorda, Hydroclathrus clathratus, Prionitis crispata, Sargassum micracanthum, and Ulva lactuca collected in Korea. Morphological and phylogenetic analyses identified the isolates as four Aspergillus spp. (A. fumigatus, A. sydowii, A. tamarii, and A. terreus), three Penicillium spp. (P. crustosum, P. jejuense, and P. rubens), and Cladosporium tenuissimum. Among them, A. fumigatus TOP-U2, A. tamarii SH-Sw5, and A. terreus GJ-Gf2 strains showed the activities of all enzymes examined (amylase, chitinase, lipase, and protease). Based on the enzymatic index (EI) values in solid media, A. terreus GJ-Gf2 and C. tenuissimum UL-Pr1 exhibited the highest amylase and lipase activities, respectively. Chitinolytic activity was only observed in A. terreus GJ-Gf2, A. tamarii SH-Sw5, and A. fumigatus TOP-U2. Penicillium crustosum UL-Cl2 and C. tenuissimum UL-Pr1 showed the highest protease activities. To the best of our knowledge, this is the first report of lipolytic and proteolytic activities in a marine-derived C. tenuissimum strain. Overall, the fungal strains isolated from the marine macroalgae in this study actively produced industrially important enzymes.

The comparison of the accuracy of temporary crowns fabricated with several 3D printers and a milling machine

  • Junsik Lee;Sungwon Ju;Jihyung Kim;Sion Hwang;Jinsoo Ahn
    • The Journal of Advanced Prosthodontics
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    • 제15권2호
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    • pp.72-79
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    • 2023
  • PURPOSE. The purpose of this in vitro study was to compare the accuracy of various 3D printers and a milling machine. MATERIALS AND METHODS. The die model was designed using CAD (Autodesk Inventor 2018 sp3). The 30 ㎛ cement space was given to the die and the ideal crown of the mandibular left first molar was designed using CAD (ExoCAD). The crowns were produced using the milling machine (Imes-icore 250i) and the 3D printers (Zenith U, Zenith D, W11) and they were divided into four groups. In all groups, the interior of each crown was scanned (Identica blue) and superimposed (Geomagic Control X) with the previously designed die. The difference between the die and the actual crown was measured at specific points. The Kruskal-Wallis test, the Mann-Whitney test, and Bonferroni's method were performed with a statistical analysis software (P < .008 in inter-group comparison P < .001 in intra-group comparison). RESULTS. In all groups, the center of the occlusal area and the anti-rotational dimple area showed significantly greater difference and the marginal area showed the smallest difference comparatively. The mean value of the difference in each area and the sum of the differences were higher in order of W11, Imes-icore 250i, Zenith D, and Zenith U. CONCLUSION. The digital light processing (DLP) method shows higher accuracy compared to the sereolithography (SLA) method using the same resin material.

고유전율 필드 플레이트를 적용한 β-Ga2O3 쇼트키 장벽 다이오드 (Vertical β-Ga2O3 Schottky Barrier Diodes with High-κ Dielectric Field Plate)

  • 박세림;이태희;김희철;김민영;문수영;이희재;변동욱;이건희;구상모
    • 한국전기전자재료학회논문지
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    • 제36권3호
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    • pp.298-302
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    • 2023
  • In this paper, we discussed the effect of field plate dielectric materials such as silicon dioxide (SiO2), aluminum oxide (Al2O3), and hafnium oxide (HfO2) on the breakdown characteristics of β-Ga2O3 Schottky barrier diodes (SBDs). The breakdown voltage (BV) of the SBDs with a field plate was higher than that of SBDs without a field plate. The higher dielectric constant of HfO2 contributed to the superior reduction in electric field concentration at the Schottky junction edge from 5.4 to 2.4 MV/cm. The SBDs with HfO2 field plate showed the highest BV of 720 V, and constant specific on-resistance (Ron,sp) of 5.6 mΩ·cm2, resulting in the highest Baliga's figure-of-merit (BFOM) of 92.0 MW/cm2. We also investigated the effect of dielectric thickness and field plate length on BV.

Phytochemical and Anti-Termite Efficiency Study of Guibourtia tessmanii (harms) J. Léonard (Kévazingo) Bark Extracts from Gabon

  • NKOGO, Ley-Fleury ELLA;BOPENGA, Christ Stone Arnaud BOPENGA;NGOHANG, Franck Estime;MENGOME, Line Edwige;ANGONE, Sophie ABOUGHE;ENGONGA, Prosper EDOU
    • Journal of the Korean Wood Science and Technology
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    • 제50권2호
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    • pp.113-125
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    • 2022
  • This study aimed to explore the biodiversity of chemical compounds found in the bark of Guibourtia tessmannii from Gabon, commonly called Kévazingo, and evaluate their anti-termite activity to determine their potential values as a source of development of anti-termite products that can be valued in the fields of fine chemicals and wood preservation. Extraction of G. tessmannii bark powders was carried out using the cold maceration method with trichloroethylene, acetone, ethanol, and water. Phytochemical screening made it possible to highlight groups of chemical families present in the extracts. Anti-termite activity was tested on the wild termites "Cubitermes sp" of the genus Isoptera. The yield of the extracts were 17.11% for the buttress and 13.42% for the height at 6 m. Phytochemical tests revealed that alkaloids, polyphenols, sterols, tannins, reducing compounds, flavonoids, saponins, and anthraquinones were present in the extracts. Results of anti-termite activity indicated that anti-termite activity varied with the different parts of the bark studied, extraction solvent, and concentration (50/50) and (25/75) of the extracts used. The extracts at 50/50 concentration showed a slightly better anti-termite activity compared to the 25/75 concentration. In addition, the buttress Kévazingo or buttress showed the strongest anti-termite activity for the aqueous extract with a survival rate of 0% after 2 days.

Anti-inflammatory activity of 6-O-phospho-7-hydroxycoumarin in LPS-induced RAW 264.7 cells

  • Hong, Hyehyun;Park, Tae-Jin;Jang, Sungchan;Kim, Min-Seon;Park, Jin-Soo;Chi, Won-Jae;Kim, Seung-Young
    • Journal of Applied Biological Chemistry
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    • 제65권1호
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    • pp.33-41
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    • 2022
  • Esculetin (also known as 6, 7-dihydroxycoumarin) a type of coumarin, has been exhibited anti-inflammatory and anti-aging effects. Biorenovation is the microbe-mediated enhancement of biological efficacies and structurally diversified compounds relative to their substrate compounds. The production of different kinds of esculetin derivatives using Bacillus sp. JD3-7 and their effects on lipopolysaccharide (LPS)-triggered inflammatory response in RAW 26.7 cells were assessed. One of the biorenovation products, identified as esculetin 6-O-phosphate (ESP), at concentrations of 1.25, 2.5, and 5 μM inhibited the LPS-stimulated production of inflammation markers of nitric oxide synthase 2 and cyclooxygenase 2 as well as their respective enzymatic reaction products of nitric oxide and prostaglandin E2 in the order of increasing concentrations (1.25, 2.5, and 5 μM). Additionally, ESP treatment suppressed the LPS-stimulated secretion of pro-inflammatory cytokines of interleukin (IL)-1β, IL-6, and tumor necrosis factor- α. Furthermore, these anti-inflammatory effect of ESP was associated with the downregulation of mitogen-activated protein kinase signaling, that is, extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase signaling pathways. This study would therefore provide interesting insights into the biorenovation-assisted generation of a novel anti-inflammatory compound. ESP may be used to develop treatments for inflammatory disorders.

파킨슨병 동물 모델을 이용한 신정격 사암침법의 도파민성 신경세포 보호 효과 연구 (Effect of Kidney Tonification of Saam Acupuncture in Parkinson's Disease Mouse Model)

  • 김승태;이상협;김보경
    • Korean Journal of Acupuncture
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    • 제39권1호
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    • pp.8-15
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    • 2022
  • Objectives : Saam acupuncture is one of the indigenous therapeutic modalities in traditional Korean medicine. In this study, the neuroprotective effect of Saam acupuncture of kidney tonification was investigated using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. Methods : Twelve-week-old male C57BL/6 mice were intraperitoneally administered with 30 mg/kg of MPTP at 24-h intervals for 5 days and acupuncture stimulation at LU8, KI7, SP3 and KI3 was performed once a day for 12 days from the first MPTP injection. The pole test and the rotarod test were performed to evaluate motor function, and dopaminergic neuronal survival in the substantia nigra (SN) and striatum was evaluated using tyrosine-hydroxylase immunohistochemistry. Results : MPTP administration caused behavioral impairment and dopaminergic neuronal death in the nigrostriatal pathway. Whereas the Saam acupuncture treatment alleviated the MPTP-induced motor dysfunction and dopaminergic neuronal death in the SN and striatum. Conclusions : Saam acupuncture of kidney tonification can alleviate the MPTP-induced motor dysfunction and dopaminergic neuronal death in the nigrostriatal pathway, suggesting a possible role for acupuncture in the treatment of Parkinson's disease.