• Title/Summary/Keyword: cell infection

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Susceptibility to FAS induced apoptosis in mycoplasma contaminated HCE cells (Mycoplasma가 오염된 배양 각막상피 세포의 FAS 유도 세포고사의 민감성)

  • Kim, Jai-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.12 no.2
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    • pp.79-86
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    • 2007
  • The aim of the present study was to determine mechanisms of corneal epithelial cell apoptosis in vitro following exposure to anti-FAS and anti-FAS ligand antibody and during infection with mycoplasma sp.. A cultured human corneal epithelial(HCE) cell line was treated with anti-FAS antibody or anti-FAS ligand antibody for 2 and 4 days. The original cell line was found to be contaminated by mycoplasma removal agent(MRA) was used to eliminate the bacterium from the cell line. MRA($0.5{\mu}{\ell}$ tissue culture medium) was added to the cell line and incubated for 1 week. The cell line underwent multiple passages in media not contaminating MRA and cells were grown to 50-80% confluency on coverslips and stained using the Hoechst stain provided in the kit to ensure mycoplasma removal. Apoptosis experiments were performed before and after mycoplasma removal. The apoptotic index of anti-FAS and anti-FAS ligand antibody on mycoplasma contaminated cell line was studied using Hoechst 33342 staining and Annexin V-FITC and Propidium Iodide Staining. In conclusion, anti-FAS antibody induces apoptosis in HCE cells in a time and concentration-dependent mechanism. Cell lines contaminated with mycoplasma have an incresed susceptibility to FAS induced apoptosis.

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Temporal Transcriptome Analysis of SARS-CoV-2-Infected Lung and Spleen in Human ACE2-Transgenic Mice

  • Jung Ah, Kim;Sung-Hee, Kim;Jung Seon, Seo;Hyuna, Noh;Haengdueng, Jeong;Jiseon, Kim;Donghun, Jeon;Jeong Jin, Kim;Dain, On;Suhyeon, Yoon;Sang Gyu, Lee;Youn Woo, Lee;Hui Jeong, Jang;In Ho, Park;Jooyeon, Oh;Sang-Hyuk, Seok;Yu Jin, Lee;Seung-Min, Hong;Se-Hee, An;Joon-Yong, Bae;Jung-ah, Choi;Seo Yeon, Kim;Young Been, Kim;Ji-Yeon, Hwang;Hyo-Jung, Lee;Hong Bin, Kim;Dae Gwin, Jeong;Daesub, Song;Manki, Song;Man-Seong, Park;Kang-Seuk, Choi;Jun Won, Park;Jun-Won, Yun;Jeon-Soo, Shin;Ho-Young, Lee;Jun-Young, Seo;Ki Taek, Nam;Heon Yung, Gee;Je Kyung, Seong
    • Molecules and Cells
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    • v.45 no.12
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    • pp.896-910
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    • 2022
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.

Effect of Postpneumonectomy Empyema on Survival of Patients with Bronchogenic Carcinoma -4 Cases Report- (폐암환자의 전폐절제술후 발생한 농흉 치험 4예)

  • 김종호
    • Journal of Chest Surgery
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    • v.13 no.3
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    • pp.285-291
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    • 1980
  • Post pneumonectomy empyema either with bronchopleural fistula or without bronchopleural fistula is an infrequent postoperative complication, but very serious and critical problem. But it is of some interest that the development of a postoperative empyema following resection for carcinoma of the lung might have a favorable effect on the survival of patients in recent speculation of the literature. We have experienced 4 cases of postoperative empyema following pneumonectomy for carcinoma of the lung at department of chest surgery, Yon Sei University, medical college during 11 years from Jan. 1968 to June 1980. Histologically, 3 cases were demonstrated squamous cell carcinoma except one oat cell carcinoma. Onset of postoperative empyema occurred over a wide range of time, from as early as the 5th postoperative day to insidious onset 6 months after pneumonectomy. The most common organisms isolated from the empyema cavities were staphylococcus aureus, pseudomonas aeruginosa and gram negative bacilli. All cases had a large number of organisms and more infections but not single infection. 2 out of 4 cases are treated with open pleural window drainage and irrigation with antibiotic`s solution 2 or 3 times per week by this time and postoperative general course is not eventful. One is alive to 2 years 3 months, another is alive to 8 years 11 months until now. And 2 out of 4 patients is survived over 4 years 10 months. Analysis of postoperative empyema complicating pneumonectomy for bronchogenic carcinoma revealed an increase in 4 year 10 months survival [50%].

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Incidence and Causes of Sub-Clinical Mastitis in Dairy Cows on Smallholder and Large Scale Farms in Tropical Areas of Tanzania

  • Shem, M.N.;Malole, J.M.L.;Machangu, R.;Kurwijila, L.R.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.372-377
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    • 2001
  • A study on the prevalence and causes of sub-clinical mastitis was conducted on ten smallholder and large-scale dairy farms in Morogoro urban and peri-urban areas. A total of 65 lactating cows were screened using the California Mastitis Test (CMT). Confirmatory tests used included; the direct microscopic somatic cell count (DMSCC), culture, bacteriological and biochemical tests. Structured questionnaires were used to collect information on management aspects. Results showed 62% and 4% cows as sub- clinical and clinical mastitis cases respectively. Levels of infection were higher on smallholder farms (75%) than on large-scale farms (25%). All tested cows had high cell counts (>500,000) per ml of milk. Incidences of mastitis were significantly (p<0.05) related to milking practices. The dominant bacterial isolates in the same order were Staphylococcus aureus, Streptococcus spp, and Escherichia coli. Other organisms isolated included Pseudomonas spp and Klebsiella spp. It was concluded that the high rates of sub-clinical mastitis in the research area were mainly due to poor management and unhygienic milking practices.

Characterizing of Rice Blast Lesion Mimic

  • Lee, Joo-Hee;Jaw, Nam-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.68.1-68
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    • 2003
  • When plants are infected by plant pathogens, typical disease symptom termed lesion, appears in compatible interaction. Whereas, in incompatible interactions, only small speck of lesions are visible on the leaf surfaces. Hypersensitive response (HR) of plant which is the result of infection by incompatible pathogens, is a well known defense response inducing rapid cell death resulting in complete resistance. However, some rice mutants show spontaneous disease symptoms during the growth stages without interaction with pathogens. We investigated the spontaneous cell death mutant called Blast Lesion Mimic(BLM) generated by EMS mutation, on the relationship with the hypersensitive response as well as resistant characteristics. Accumulation of phenolic compounds were detected around the lesions as lesions develop on leaf surface. Activation of PR gene was detected before the lesion appeared, and that result indicates the defense-related response are started earlier than lesion formation. The BLM mutant showed resistant response to inoculation of Magnaporthe grisea KJ201 with which the wild type Hwacheong is totally susceptible. Informations on the formation of spontaneous lesions and detail analysis of lesion mimic mutants and related genes are very limited to date. It is really important to understand the phenomenon of the defense-related lesion formation for developing resistant cultivar for rice blast pathogens

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Immunocytochemical Staining for p16 of Atypical Squamous Cells in Cervicovaginal Smear (자궁목질 바른표본에서 비정형 편평세포의 p16에 대한 면역세포화학염색)

  • Kim, Hwal-Woong;Lee, Jong-Sil;Lee, Jeong-Hee;Ko, Gyung-Hyuck
    • The Korean Journal of Cytopathology
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    • v.15 no.1
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    • pp.28-32
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    • 2004
  • It was reported that the main cause of intraepithelial neoplasm and squamous cell carcinoma of the uterine cervix is human papilloma virus infection, and that the expression of p16 is increased in cells infected by human papilloma virus. We performed an immunocytochemical staining for protein p16 in 17 cases of cervocovaginal smears initially diagnosed as atypical squamous cells of undetermined significance, to know whether the staining could help the differentiation of neoplastic cells from reactive atypical cells. Of 17 smears, 6 were diagnosed finally as high grade intraepithelial neoplasm or invasive squamous cell carcinoma by follow-up biopsy and smear, and 5 of the 6 were positive for p16. Three were diagnosed as koilocytosis, and one of them was weakly positive for p16. Eight were diagnosed as reactive atypical cells, and all of them were negative for p16. We thought that immunocytochemical staining of p16 in cervocovaginal smears could help the differentiation of neoplastic cells from reactive atypical cells.

Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok;Chung, Yong-Ju;Jeon, Yeong-Joong;Lee, Sung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.386-392
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    • 1999
  • A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95% of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40% of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

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Molecular Analysis of Geminigirus ORFs on Symptom Development

  • Park, Eulyong;Hyunsik Hwang;Lee, Sukchan
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.38-43
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    • 1999
  • Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

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Possible Role of Heme Oxygenase-1 and Prostaglandins in the Pathogenesis of Cerebral Malaria: Heme Oxygenase-1 Induction by Prostaglandin $D_2$ and Metabolite by a Human Astrocyte Cell Line

  • Kuesap, Jiraporn;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • v.48 no.1
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    • pp.15-21
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    • 2010
  • Astrocytes are the most abundant cells in the central nervous system that play roles in maintaining the blood-brain-barrier and in neural injury, including cerebral malaria, a severe complication of Plasmodium falciparum infection. Prostaglandin (PG) $D_2$ is abundantly produced in the brain and regulates the sleep response. Moreover, $PGD_2$ is a potential factor derived from P. falciparum within erythrocytes. Heme oxygenase-1 (HO-1) is catalyzing enzyme in heme breakdown process to release iron, carbon monoxide, and biliverdin/bilirubin, and may influence iron supply to the P. falciparum parasites. Here, we showed that treatment of a human astrocyte cell line, CCF-STTG1, with $PGD_2$ significantly increased the expression levels of HO-1 mRNA by RT-PCR. Western blot analysis showed that $PGD_2$ treatment increased the level of HO-1 protein, in a dose- and time-dependent manner. Thus, $PGD_2$ may be involved in the pathogenesis of cerebral malaria by inducing HO-1 expression in malaria patients.

New trends of vaccine development: Recombinant vaccinia viruses (expression vectors) as vaccines (Vaccine개발(開發)의 새로운 동향(動向) : Vaccinia virus를 발견(發見) vector로 이용하는 재조합(再組合) 생(生)vaccine의 작성(作成))

  • Kim, Uh-ho
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.407-416
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    • 1989
  • The prospect of live vaccines consisting of genetically modified vaccinia virus expressing foreign genes is exciting, but important issues concerning safety and efficacy need to resolved. Vaccinia virus (VV) is an efficient expression vector with broad host range infectivity and large DNA capacity. This vector has been particularly useful for identifying target antigens for humoral and cell-mediated immunity. The WHO smallpox eradication program, involving the extensive use of VV vaccines, resulted in the late 1970s in the elimination of one of the world's most feared diseases. This achievement is a triumph for preventive medicine and for international collaboration in public health. In 1980, WHO recommended that the routine use of smallpox vaccine should be stopped. Against this background, the prospect of li ve vaccines consisting of genetically modified VV expressing foreign antigens arising from the work of Moss, and Paoletti and their colleagues in 1982 has been greeted with enthusiasm. These investigators have shown that genes coding for immunogenic proteins can be inserted into VV DNA without impairing the ability of the virus to grow in cell culture. Moreover experimental animals infected with VV recombinants containing genes coding for a variety of immunizing proteins have been shown to be protected against challenge infection with the corresponding infectious agent. In this communication, I describe current progress in the construction of a novel plasmid vector that facilitate the insertion and expression of foreign genes in VV as well as the selection of recombinants.

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