• Title/Summary/Keyword: Protein stain

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Complete genome sequence of Spirosoma aerolatum KACC 17939T, a bacterium related to the DNA repair (DNA 복원에 관련된 박테리아 Spirosoma aerolatum KACC 17939T의 완전한 게놈 서열)

  • Kim, Dong-Uk;Kim, Ju-Young;Kim, Su Jeong;Kim, Min Ji;Lee, Ju Yeon;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.230-232
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    • 2017
  • A Gram-stain-negative, yellow-pigmented bacterial strain, designated Spirosoma aerolatum KACC $17939^T$, was isolated from a biofilm of car air conditioner collected in Republic of Korea. In this study, we report the complete genome sequence of a bacterium Spirosoma aerolatum KACC $17939^T$ obtained using the PacBio RS II platform. The genome comprised of 7,959,595 bp with the G + C content of 48.3%, the genome included 6,640 genes were predicted, among them, 6,471 genes are protein-coding genes.

A Comparative Study of Eejin-tang, Hyangsaeejin-tang and Naeso-san Extracts on Indomethacin-induced Gastric Mucosal Lesions in Mice (Indomethacin으로 유발된 생쥐의 위점막 손상에 대한 이진탕(二陳湯), 향사이진탕(香砂二陳湯) 및 내소산(內消散)의 비교연구)

  • Park, Jung-Han;Baik, Tai-Hyeun
    • The Journal of Internal Korean Medicine
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    • v.34 no.4
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    • pp.412-427
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    • 2013
  • Objectives : This study was carried out to compare the effects of Eejin-tang, Hyangsaeejin-tang and Naeso-san extracts on indomethacin-induced gastric mucosal lesions in mice. Methods : Experimental mice were divided into six groups. The normal group had no gastro-inflammation. In the control group, gastro-inflammation was elicited by indomethacin. Misoprostol, Eejin-tang, Hyangsaeejin-tang and Naeso-san group were those in which misoprostol, Eejin-tang extract, Hyangsaeejin-tang extract and Naeso-san extract were administered after gastro-inflammation is elicited. This study examined the anti-inflammation effects and distribution of mucus secreting cells, zonula occludin-1 (ZO-1), heat shock protein (HSP) 70, periodic acid-schiff reaction stain (PAS), peanut agglutinin (PNA), cyclooxygenase-1 (COX-1), 5-bromo-2'-deoxyuridine (BrdU), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) p65, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Results : 1. The hemorrhagic erosion and damaged mucus secreting cell, the positive reaction HSP70 increased in the control group, but decreased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. 2. The positive reaction of ZO-1, PAS, PNA, COX-1 and BrdU decreased in the control group, but increased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. 3. The positive reaction of NF-${\kappa}B$ p65, iNOS and COX-2 increased in the control group, but decreased in the Eejin-tang, Hyangsaeejin-tang and Naeso-san groups. Conclusions : Among the three extracts, the effects were in the order of Naeso-san, Hyangsaeejin-tang and Eejin-tang group, Naeso-san being the most effective.

A Case of Secondary Renal Amyloidosis in Children with Juvenile Rheumatoid Arthritis (소아기 류마티스 관절염 환아에서 발생한 이차성 신유전분증 1 례)

  • Moon Jae-Hoon;Lee Suk-Jin;Kang Mi-Seon;Chung Woo-Yeong
    • Childhood Kidney Diseases
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    • v.6 no.2
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    • pp.243-250
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    • 2002
  • Amyloidosis comprises a diverse group of systemic and local diseases characterized by organ involvement by the extracellular deposition of fibrils composed of subunits of a variety of normal serum proteins. Secondary amyloidosis is caused by the deposition of amyloid A(AA) protein in chronic inflammatory disease. Juvenile rheumatoid arthritis(JRA) has been known to be the most common cause of secondary amyloidosis. We experienced one case of secondary renal amyloidosis in a 12-year-old girl who had suffered from JRA for several years who had visited our renal clinic to evaluate the proteinuria with microscopic hematuria which was detected by chance at school urine screening examination. Apple green birefringence was observed under polarized light with Congo red stain at)d characteristic electron microscopic findings was also noted in renal tissues which was obtained by percutaneous renal biopsy. In our knowledge, this is the first case report of secondary renal amyloidosis developed in pediatric age in Korea.

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Rapid detection of Theileria sergenti by polymerase chain reaction (중합효소연쇄반응을 이용한 Theileria sergenti의 신속한 검출)

  • 최은진;강승원
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.111-118
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    • 1997
  • Four separate pairs of oligonucleotide primers within the coding region in a T sergenti 33-kDa surface protein gene were selected to detect T. sergenti by PCR. The specificity of PCR-amplified DNA was examined by digestion with restriction enzyme 3nd Southern blot hybridization using T. sergenti p33 DNA probe. PCR appears to be specific for T. sergenti, without detectable signals from uninfected erythrocytes, uninfected bovine leukocytes and other hemoparasites, including A. morginnle and 3. ouata. Although 46 of 71 specimens (64.8%) from grazing cattle were microscopically positive. PCR in this study showed that 64 specimens (88.7%) were positive. Therefore, PCR proves a useful diagnostic tool for detecting T sergenti-infected cattle. In addition, it is also revealed that PCR was significantly more sensitive than traditional microscopic examination using Giemsa's stain.

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Resveratrol-Enriched Rice Down-Regulates Melanin Synthesis in UVB-Induced Guinea Pigs Epidermal Skin Tissue

  • Lee, Taek Hwan;Seo, Jae Ok;Do, Moon Ho;Ji, Eunhee;Baek, So-Hyeon;Kim, Sun Yeou
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.431-437
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    • 2014
  • Synthetic compounds that are used in the clinic to regulate skin hyperpigmentation, such as arbutin, hydroquinone, and kojic acid, are only moderately effective. But, their use is limited by side effects. As part of an effort to overcome the limitations, we developed resveratrol-enriched rice (RR) using genetic engineering technique. Each of resveratrol and rice has been reported to produce anti-melanogenic effects. Therefore, we hypothesized that RR would show more anti-melanogenic effects than those of resveratrol or rice alone. Anti-melanogenic effect of RR was done by using melan-a mouse melanocytes. The depigmenting efficacy was then observed following topical application of the RR to UVB-stimulated hyperpigmented dorsal skin of guinea pigs. Treatment with RR extract resulted a $21.4{\pm}0.7%$ decrease in tyrosinase expression at melan-a cells. Colorimetric analysis showed a significantly lower depigmenting value by day 9 following treatment with RR in UVB-irradiated guinea pigs the dorsal skin (p<0.01), indicating that RR produced a depigmentation effect. By staining with Fontana-Masson stain, we found that the RR-treated group had more effect histopathologically in epidermal melanin production than resveratrol or rice alone-treated group. RR was associated with reduction in the levels of microphthalmia-associated transcription factor (MITF), and downregulation of tyrosinase and tyrosinase-related protein (TRP-2) expression, leading to inhibit epidermal melanin production by western blot analysis. This study suggests that the resveratrol-enriched rice may be a promising candidate in regulating skin pigmentation with UVB exposure.

A STUDY ON THE HISTOLOGICAL CHANGES AND THE DISTRIBUTION OF LANGERHANS CELLS OF THE ALVEOLAR MUCOSA IN DENTURE AND NONDENTURE WEARERS (무치악 환자에서 의치장착에 따른 치조점막의 조직학적 변화 및 Langerhans세포의 분포에 관한 연구)

  • Lee, Hyeog;Lee, Ho-Yong
    • The Journal of Korean Academy of Prosthodontics
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    • v.29 no.2
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    • pp.211-223
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    • 1991
  • This study was aimed to observe the histological changes of the edentulous and denture wearing alveolar ridge mucosa. The distribution of Langerhans cells was also observed to investigate the mucosal immune response by denture wearing. The mucosal tissues were obtained from of 12 cases of edentulous nondenture wearers(NDW), 7 cases of denture wearers(DW), and 12 cases of flabby tissues(FT). For the identification of Langerhans cells of the mucosal epithelia, the immunohistochemical stain for S-100 protein was applied. The results were as follows : 1. 7 cases among 12 cases of NDW showed hyperkeratosis, and 5 cases were covered by parakeratosis, whereas 3 cases among 7 cases of DW showed hyperkeratosis, and 4 cases showed parakeratosis on the mucosal epithelium. All cases of both DW and NDW demonstrated epithelial hyperplasia, except. 2 cases of DW, which showed epithelial atrophy. The content of glycogen in the epithelial layer showed the decrease in the group of DW. 2. Both NDW and DW showed the infiltration of chronic inflammatory cells. The collagen fibers tended to be arranged densely and irregularly in cases of denture wearing period more than 10 years. 3. FT showed variable epithelial changes from epithelial atrophy to marked hyperplasia, and the pattern of keratinization was also variable. The collagen fibers tended to be arranged irregularly. 4. The distribution of Langerhans cells showed the increase of 1.84-1.96 times in the group of DW compared with NDW group.

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Characteristics of contact and distance osteogenesis around modified implant surfaces in rabbit tibiae

  • Choi, Jung-Yoo;Sim, Jae-Hyuk;Yeo, In-Sung Luke
    • Journal of Periodontal and Implant Science
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    • v.47 no.3
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    • pp.182-192
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    • 2017
  • Purpose: Contact and distance osteogenesis occur around all endosseous dental implants. However, the mechanisms underlying these processes have not been fully elucidated. We hypothesized that these processes occur independently of each other. To test this, we used titanium (Ti) tubes to physically separate contact and distance osteogenesis, thus allowing contact osteogenesis to be measured in the absence of possible triggers from distance osteogenesis. Methods: Sandblasted and acid-etched (SLA) and modified SLA (modSLA) implants were used. Both types had been sandblasted with large grit and then etched with acid. The modSLA implants then underwent additional treatment to increase hydrophilicity. The implants were implanted into rabbit tibiae, and half were implanted within Ti tubes. The bone-to-implant contact (BIC) ratio was calculated for each implant. Immunohistochemical analyses of bone morphogenetic protein (BMP)-2 expression and new bone formation (Masson trichrome stain) were performed. Results: The implants outside of Ti tubes were associated with good bone formation along the implant surface. Implantation within a Ti tube significantly reduced the BIC ratio (P<0.001). Compared with the modSLA implants, the SLA implants were associated with significantly higher BIC ratios, regardless of the presence or absence of Ti tubes (P=0.043). In the absence of Ti tubes, the bone adjacent to the implant had areas of new bone formation that expressed BMP-2 at high levels. Conclusions: This study disproved the null hypothesis and suggested that contact osteogenesis is initiated by signals from the old bone that undergoes distance osteogenesis after drilling. This signal may be BMP-2.

Secretion of Ferritin Protein of Periserrula leucophyryna in Bacillus subtilis and Its Feed Efficiency (고초균에서 흰이빨참갯지렁이 페리틴 단백질의 분비 및 사료 효율성)

  • Choi, Jang Won
    • KSBB Journal
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    • v.31 no.2
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    • pp.105-112
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    • 2016
  • Ferritin is known to regulate iron metabolism and maintain iron in a variety of the eukaryotic organisms. The region encoding the mature ferritin (0.47 kb, H-type) of Periserrula leucophryna was amplified using the designed primers including restriction enzyme site and termination codon and subcloned in frame to the pRBAS secretion vector containing the signal sequence, RBS, and promoter of amylase gene (E. coli-Bacillus shuttle vector), resulting in recombinant pRBAS-PLF vector. Recombinant ferritin (18 kDa) was correctly processed and secreted from Bacillus subtilis LKS strain harboring the pRBAS-PLF vector and quantitatively analyzed by SDS-PAGE and western blot, respectively. Secretion of the ferritin was optimized by culture conditions (host, medium, temperature, nitrogen source) in 3 L batch culture and 5 L jar fermenter. Finally. the ferritin was largely produced using 50 L fermenter as the following conditions; at $30^{\circ}C$, 150 rpm, 1 vvm in Bacillus subtilis LKS using PY medium. The secreted ferritin was maximally measured (approximately 177.6 ug/ml) when the cell density reached to 14.4 at $OD_{600}$ (20 h incubation). The iron binding activity was confirmed by Perls' staining in 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in the culture broth after secretion. Biologically, the culture broth and powder type containing ferritin were tested for possibility as feed additive in chicken broiler. As a result, the ferritin stimulated the growth of chick broil and improved feed efficiency and production index.

High-Level Expression of an Aspergillus niger Endo-$\beta$-1,4-Glucanase in Pichia pastoris Through Gene Codon Optimization and Synthesis

  • Zhao, Shumiao;Huang, Jun;Zhang, Changyi;Deng, Ling;Hu, Nan;Liang, Yunxiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.467-473
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    • 2010
  • To improve the expression efficiency of recombinant endo-$\beta$-1,4-glucanase in P. pastoris, the endo-$\beta$-1,4-glucanase (egI) gene from Aspergillus niger was synthesized using optimized codons. Fourteen pairs of oligonucleotides with 15 bp overlap were designed and the full-length syn-egI gene was generated by two-step PCR-based DNA synthesis. In the synthesized endo-$\beta$-1,4-glucanase gene syn-egI, 193 nucleotides were changed, and the G+C content was decreased from 54% to 44.2%. The syn-egI gene was inserted into pPIC9K and transformed into P. pastoris GS115 by electroporation. The enzyme activity of recombinant P. pastoris stain 2-7# reached 20.3 U/ml with 1% barley $\beta$-glucan and 3.3 U/ml with 1% carboxymethylcellulose (CMC) as substrates in shake flasks versus 1,270.3 U/ml and 220.7 U/ml for the same substrates in 50-1 fermentors. The molecular mass of the recombinant protein was approximately 40 kDa as determined by SDS-PAGE analysis, the optimal temperature for recombinant enzyme activity was $70^{\circ}C$, and the optimal pH was 5.0 when CMC was used as the substrate.

Induction of Apoptosis by Tosyl-JM3 in HL-60 cells

  • Kim Kun-Jung;Ju Sung-Min;Lee Chai-Ho;Kim Won-Sin;Yun Yong-Gab;Jeong Han-Sol;Kim Sung-Hoon;Park Sung-Joo;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1370-1374
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    • 2005
  • The Tosyl-JM3 (TJM3) is a modified compound from one of 1,2,3,4-Tetra- hydroisoquinoline (THIQ) derivatives. The THIQs include potent cytotoxic agents that display a range of anti-tumor activities, antimicrobial activity, and other biological properties. In this study, we investigated the effect of TJM3 on the cytotoxicity, induction of apoptosis in human promyelocytic leukemia cells (HL-60 cells). TJM3 showed a significant cytotoxic activity in HL-60 cells (IC50 = approximately $60{\mu}g/m{\ell}$) after a 24 hr incubation. Treatment of HL-60 cells with TJM3 exhibited several features of apoptosis, including formation of DNA ladders in agarose gel electrophoresis, morphological changes of HL-60 cells with DAPI stain. Here we observed that TJM3 caused a decrease of procaspase-3 protein. Further molecular analysis demonstrated that TJM3 led to cleavage of poly(ADP-ribose) polymerase (PARP) by western blot and increase of hypodiploid (Sub-G1) population in the flow cytometric analysis. In conclusion, these above results indicate that TJM3 dramatically suppresses HL-60 cell growth and induces apoptosis. These data may support a possibility for the use of TJM3 in the prevention and treatment of leukemia.