• Restorative Dentistry and Endodontics
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• v.19 no.2
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• pp.485-498
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• 1994

Porphyromonas endodontalis is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections and this microorganism possesses a potential for pathogenicity. The purpose of this study was to compare the membrane components of Porphyromonas endodontalis and Porphyromonas gingivalis and to study the immune reaction patterns of Porphyromonas endodontalis with patients with periapical lesion. Porphyromonas endodontalis (ATCC 35406), Porphyromonas gingivals serotypea (381), serotype b(W50), serotype c(A7A1-28) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells and human sera were obtained from patients and dental students. Indirect immunofluorescence method was used to study on the cross reaction between Porphyromonas endodontalis and Porphyromonas gingivalis serotype a, b, c antigen. Total membrane protein profiles of Porphyromonas endodontalis antigen were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the reactivity of antigenic components of Porphyromonas endodontalis against sera of patients and rabbit anti-Porphyromonas endodontalis antisera were assessed by Immunoblotting method. The following results were obtained : 1. Antigens of Porphyromonas endodontalis has multiple antigenic components, and both patients with periapical lesion and normal healthy individual showed immune response to this. 2. Patients group and healthy individual group showed a diversity of immune reaction pattern but they showed immune response against 43kd protein. 3. Patients with periapical lesion showed more diverse immune response than healthy individual and in some patients, much more bands appeared to lower molecular weight protein. 4. According to indirect immunofluorescence and Immunoblotting study, Porphyromonas endodontalis did not share common antigen with Porphyromonas gingivalis serotype a, b, c.

• KSBB Journal
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• v.5 no.3
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• pp.279-291
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• 1990

Partial hydrolysed and deacetylated chitin, CHITA and CHITB as supports of immobilized enzyme were obtained by treatment of acid and base respectively. Glutaraldehyde, bifunctional reagent, was employed for crosslinking between $\beta$-glucosidase and support. Immobilized enzyme activities of CHITA-Gase and CHITB-Gase were determined with the reaction of p-nitrophenol-$\beta$-D-glucopyranoside(PNG) in batch reactor, CSTR and PFR. Their optimum temperature, pH and enzymatic characteristics including Km and Vmax values were observed with variation of the flow rates. Mass transfer coefficient(h), effectiveness factor(η), deactivation rate(kd ) of two immobilized enzymes were also examined to compare efficiency of reactors.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.953-964
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• 1999

본 연구는 세 종류의 Porphyromonas gingivalis(Pg) antigen의 IgG subclass associated recognition을 평가하기 위해 수행했었다. 총 35명의 조기발병형치주질환자중, Pg381에 대한 IgG2항체의 증가를 보이는 5명이 급속진 행형 치주질환자, IgG4의 증가를 보이는 6명의 환자(국소유년형 치주질환자 2명과 급속진행 형 치주질환자 4명), IgG2+4의 증가를 보이는 2명의 급속진행형 치주질환자 그리고 IgG1+2+4의 증가를 보이는 8명의 환자(국소 유년형 치주질환자 2명과 급속진행형 치주질환자 6명)으로 구성된 21명의 환자를 dot immunoblot analysis를 위해 선택했다. 실험에 사용된 정제된 항원은 Pg381에서 추출한 43-kd fimbrilin protein과 lipoplysaccharide(LPS), Pg A7A1-28(ATCC 53977)에서 추출한 capsularpolysaccharide(CPS)였다. Immunoblotting pattern은 IgG4 antibody가 fimbrial antigen에 강력히 반응함을 보여주었다. Fimbriae에 잘 반응하는 몇몇의 IgG4 antibody역시 antigen에 대해 양성반응을 보였다. 대조적으로 IgG2는 CPS antigen을 일차적으로 인식했다. 전부는 아니지만 대부분의 경우, single이나 group화된 IgG subclass는 모두 LPS antigen을 인식하지 못했다. 같은 group에서 염색강도의 개인적인 차이는 증명되었다. 이런 결과는 조기발 병형 치주질환에서 Pg의 fimbriae와 CPS가 immunodominant antigen이 될 수 있음을 제시한다. 더욱이 IgG subclass antibody가 이런 Pg의 immunoglobulin antigen을 선택적으로 인식함을 알았고, 이는 조기발병형치주질환의 병리에 immunodominant antigen과 함께 IgG의 기능적인 역할을 고려해야 함을 제시한다.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.4
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• pp.377-384
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• 1997

To investigate the properties of ryanodine binding sites of the bird skeletal SR vesicles, SDS PAGE, purification of RyR, and $[^3H]ryanodine$ binding study were carried out in the SR vesicles prepared from the chicken pectoral muscle. The chicken SR vesicles have two high molecular weight (HMW) protein bands as in eel SR vesicles on SDS PAGE. The HMW bands on SDS PAGE were found in the $[^3H]ryanodine$ peak fraction $(Fr_{3-5})$ obtained from the purification step of the ryanodine receptor protein. Bmax and KD of the chicken $[^3H]ryanodine$ binding sites were 12.52 pmol/mg protein and 14.53 nM, respectively. Specific $[^3H]ryanodine$ binding was almost maximal at $50{\sim}100$ ${\mu}M$ $Ca^{2+}$, but was not increased by 5 mM AMP and not inhibited by high $Ca^{2+}$. Binding was significantly inhibited by $20{\sim}100$ ${\mu}M$ ruthenium red and 1 mM tetracaine, but slightly inhibited by $Mg^{2+}$. From the above results, it is suggested that chicken SR vesicles have the ryanodine binding sites to which the binding of ryanodine is almost maximal at $50{\sim}10$ ${\mu}M$ $Ca^{2+}$, is significantly inhibited by ruthenium red and tetracaine, slightly inhibited by $Mg^{2+}$, but not affected by AMP and not inhibited by high $Ca^{2+}$.

• Korean Journal Plant Pathology
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• v.12 no.1
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• pp.99-108
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• 1996

3종의 빙핵세균 Peudomonas syringae 8401, Pseudomonas fuorescens 8701, Erwinia herbicola 8701의 세포 외막으로부터 아무런 변성제도 사용치 않고 sucrose density gradient centrifugation, Sephacryl gel filtration chromatography, DEAE-cellulose ion exchange chromatography, non-denaturing buffer를 이용한 PAGE, electroelution, SDS-PAGE를 통해 빙핵활성 단백질을 고도로 정제할 수 있었다. P. suringae와 P. fluorescens에서는 각각 3종류(155 kD, 75 kD, 50 kD)의 빙핵활성 단백질이, E. herbicola에서는 155 kD를 제외한 2종류(75 kD, 50 kD)의 빙핵활성 단백질은 이 연구를 통해 처음 밝혀진 것으로 , 지금까지 보고된 빙핵활성 단백질(150 KD 이상)보다는 훨씬 작은 것이다. 이는 빙핵활성을 나타내는 단백질의 기본단위는 이 실험의 결과만에 의하면 최대 50 kD임을 시사한다. 이들 단백질은 그 유래된 세균의 종류나 또는 단백질 분자량의 크기에 관계없이 모두 -5.5~7.5$^{\circ}C$에서 물을 동결시키는 높은 빙핵활성을 갖고 있었다. 이는 지금까지 보고된 어느 정제단백질의 빙핵활성보다 높은 것이다. 정제된 단백질의 빙핵활성은 trypsin 처리에 의해 상실되었고, pH6~8범위에서는 안정하였으며, pH5이하, pH9이상에서는 활성을 상실하였다. 보존온도에 대한 영향은 3$0^{\circ}C$이상이 되면 점차 활성이 감소하는 경향을 보이다 37$^{\circ}C$이상에서는 활성이 완전히 상실되었다. 금속이온으로서 Hg\ulcorner이온과 SDS에 의해 활성이 상실되었으나 phosphatidylinositol의 첨가에 의해서는 활성이 약간 증가(-1$^{\circ}C$)하였다.

• Applied Biological Chemistry
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• v.43 no.2
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• pp.81-85
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• 2000

Screening was performed to isolate biopolymer-producing microorganisms from natural sources. The bacteriological characteristics of this strain and physicochemical properties of the biopolymer produced were investigated. The bacterial strain was identified as a Bacillus coagulans. Crude biopolymer treated with ethanol and actone was purified to fraction I and II by ion exchange chromatography and gel chromatography (Shephadex G-100 and Shepharose CL-2B). Analysis of chemical composition and various color reaction revealed that the polymer is composed of amino acids. It was confirmed that fraction II is a homopolymer of glutamic acid and fraction I is a homopolymer of lysine by analysis data of amino acid analyzer, GC and IR.

• Archives of Plastic Surgery
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• v.41 no.5
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• pp.562-570
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• 2014

Background Knee disarticulations (KD) are most commonly employed following trauma or tumor resection but represent less than 2% of all lower extremity amputations performed in the United States annually. KDs provide enhanced proprioception, a long lever arm, preservation of adductor muscle insertion, decreased metabolic cost of ambulation, and an end weight-bearing stump. The role for KDs in the setting of arterial insufficiency or overwhelming infection is less clear. The purpose of this study is to describe technique modifications and report surgical outcomes following KDs at a high-volume Limb Salvage Center. Methods A retrospective study of medical records for all patients who underwent a through-knee amputation performed by the senior author (C.E.A.) between 2004 and 2012 was completed. Medical records were reviewed to collect demographic, operative, and postoperative information for each of the patients identified. Results Between 2004 and 2012, 46 through-knee amputations for 41 patients were performed. The mean patient age was 68 and indications for surgery included infection (56%), arterial thrombosis (35%), and trauma (9%). Postoperative complications included superficial cellulitis (13%), soft tissue infection (4%), and flap ischemia (4%) necessitating one case of surgical debridement (4%) and four transfemoral amputations (9%). 9 (22%) patients went on to ambulate. Postoperative ambulation was greatest in the traumatic cohort and for patients less than 50 years of age, P<0.05. Alternatively, diabetes mellitus and infection reduced the likelihood of postoperative ambulation, P<0.01. Conclusions Knee disarticulations are a safe and effective alternative to other lower extremity amputations when clinically feasible. For patient unlikely to ambulate, a through-knee amputation maximizes ease of transfers, promotes mobility by providing a counterbalance, and eliminates the potential for knee flexion contracture with subsequent skin breakdown.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.327-337
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• 1999

The transposon TnphoA was used to generate avirulent mutants from a type A Pasteurella multocida. A suicide vector plasmid pRT733 carrying TnphoA, having the kanamycin resistant gene and harbored in Escherichia coli K-12 strain SM10(${\lambda}pir$), was mated with streptomycin resistant P. multocida P-1059 strain as recipient. This resulted in the generation of two TnphoA insertion mutants (transconjugants, tc95-a and tc95-b) which were resistant both to kanamycin ($Km^{R}$) and streptomycin ($Sm^{R}$), secreted alkaline phosphatase, and were avirulent to turkeys. Southern blot hybridization using two probes derived from internal fragments of TnphoA, confirmed the insertion of TnphoA into 12.9kb or 13.7kb DNA fragment from the EcoRV digested genomic fragments of transconjugants. The two transconjugants, tc95-a and tc95-b, were distinguishable from their parent strains by differences in ribotypes, and outer membrane protein profiles. TnphoA insertion in both transconjugants also resulted in constitutive expression of a 33Kd iron regulated outer membrane protein (IROMP). The gene encoding $Sm^{R}$ was also located within the same 12.9kb EcoRV genomic fragment from both transconjugants. Furthermore, our finding that the recipient P. multocida P-1059 $Sm^{R}$ strain and both transconjugants were avirulent to turkeys suggest that the either 12.9kb or 13.7kb genomic DNA contains the virulence gene and speculate that the presence of $Sm^{R}$ gene or TnphoA insertion may be responsible for regulating and inactivating the gene(s) encoding virulence in P. multocida.

• Korean Journal of Veterinary Research
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• v.33 no.3
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• pp.393-406
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• 1993

Antibacterial substances produced by Lactobacillus casei subsp. and Streptococcus faecium were examined for its antibacterial effects against some pathogenic bacteria. They were partially purified with ammonium sulfate precipitation, methanol-acetone extraction, G-50 gel filtration and examined its characteristics. When L casei subsp. and Str faecium were cultivated in MRS broth, stationary phase of L casei is until 24 hours and Str faecium is 20 hours. pH change of the cultured medium was both decreased after 12 hours and then constant at pH 4.5~4.6 after 28 hours. MRS broth culture fluids of L casei subsp. and Str faecium appeared the antibacterial effects by the spot-on-the-lawn method against ETEC, Sal pullorum and Sta aureus. Culture filtrates of L casei subsp. and Str faecium also appeared the antibacterial effects by the disc diffusion method. Culture filtrates of L casei sub. rhamnosus 7469 produced 0.032M of lactic acid and 0.01M of acetic acid. Str faecium 27273 also produced 0.027M of lactic acid and 0.01M of acetic acid. Protein concentrations of culture filtrates produced by L casei sub rhamnosus 7469 and Str faecium 27273 was $495{\mu}g/m{\ell}$ and $594{\mu}g/m{\ell}$, respectively. Antibacterial substances which are partially purified by ammonum sulfate precipitation, methanol-acetone extraction and G-50 gel filtration inhibit the growth of ETEC, Sal pullorum and Sta aureus. Characteristics of purified antibacterial substances was examined. Its molecular weight was about 31Kd, stabilized at $100^{\circ}C/20min.$ and some of proteolytic enzyme treatment.

• The Journal of the Korean Society for Microbiology
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• v.35 no.4
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• pp.299-308
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• 2000

Recently developed cDNA microarray or DNA chip technology allows expression monitoring of expression of hundreds and thousands of genes simultaneously and provides a format for identifying genes as well as changes in their activity. In order to search for changes in gene expression after X irradiation in HL60 cells, cDNA microarray technique was done. In this study, expression of 588 human genes (including oncogenes, tumor suppressor genes, cell cycle regulator genes, intracellular signal transduction modulator genes, apoptosis related genes, transcription factor genes, growth factors and receptor genes, cytokine genes, etc) were analyzed. For cDNA microarray analysis mRNAs were extracted from control and 8 Gy-irradiated HL60 cells. As a result the changes in expression of several genes were observed. This alteration of gene expression was confirmed by reverse transcription-polymerase chain reaction. The expression of heat shock 60 KD protein, c-jun, erythroid differentiation factor, CPP32, myeloid cell nuclear differentiation antigen, MAP kinase-activated protein kinase, interleukin-8, monocyte chemotactic peptide 1 and RANTES genes was increased, but the expression of p55CDC gene was decreased after X irradiation.