• Title/Summary/Keyword: pathogenic infection

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Codon Optimization, Soluble Expression and Purification of PE_PGRS45 Gene from Mycobacterium tuberculosis and Preparation of Its Polyclonal Antibody Protein

  • Xu, Tao;Li, Minying;Wang, Chutong;Yuan, Meili;Chang, Xianyou;Qian, Zhongqing;Li, Baiqing;Sun, Meiqun;Wang, Hongtao
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1583-1590
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    • 2021
  • Studies have demonstrated that PE_PGRS45 is constitutively expressed under various environmental conditions (such as nutrient depletion, hypoxia, and low pH) of the in vitro growth conditions examined, indicating that PE_PGRS45 protein is critical to the basic functions of Mycobacterium tuberculosis. However, there are few reports about the biochemical function and pathogenic mechanism of PE_PGRS45 protein. The fact that this M. tuberculosis gene is not easily expressed in E. coli may be mainly due to the high content of G+C and the use of unique codons. Fusion tags are indispensable tools used to improve the soluble expression of recombinant proteins and accelerate the characterization of protein structure and function. In the present study, His6, Trx, and His6-MBP were used as fusion tags, but only MBP-PE_PGRS45 was expressed solubly. The purification using His6-MBP tag-specific binding to the Ni column was easy to separate after the tag cleavage. We used the purified PE_PGRS45 to immunize New Zealand rabbits and obtained anti-PE_PGRS45 serum. We found that the titer of polyclonal antibodies against PE_PGR45 was higher than 1:256000. The result shows that purified PE_PGRS45 can induce New Zealand rabbits to produce high-titer antibodies. In conclusion, the recombinant protein PE_PGRS45 was successfully expressed in E. coli and specific antiserum was prepared, which will be followed by further evaluation of these specific antigens to develop highly sensitive and specific diagnostic tests for tuberculosis.

A Comparative Study on Hydrolase Activities in Acanthamoeba culbeytconi and A. roureba (Acanthamoebaculbertsoni와 A. royreba의 가수분해 효소 활성도의 비교 연구)

  • 김용규;김태우
    • Parasites, Hosts and Diseases
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    • v.26 no.2
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    • pp.95-106
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    • 1988
  • Specific or non-specific cytolytic processes of free-living amoebae causing meningoencephalitls have been emphasized and the cytolytic ability related to hydrolases in Entantoeba sp. and Naegleria sp. has also been reported since the latter half of 1970's. However, no information on hydrolase activities in Acanthamoeba sp. is available. Hydrolases in Acanthamoeba culbertsoni, a pathogenic species of free-living amoebae, were assayed and compared with those in a non-pathogenic species, A. royreba. Pathogenicity of these two species was confirmed through experimental infection to BALB/c mice. Hydrolase activities and cytotoxic effects between pathogenic and non.pathogenic species were compared in the trophozoites cultured in CGV media and in CHO cell line, respectively. The results are summarized as follows: 1. The mice infected with A. culbertseni were all dead 15 days after nasal inoculation, and the mean survival time was 8.5 days. Also the mice infected with this pathogenic species manifested typical meningoencephalitis, whereas the mice infected with A. royreba did not. 2. Hydrolases detected both in the cell extracts and culture media were acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$, neutral proteinase and acid proteinase, all of which were detected with remarkably higher rate in A. culbertsoni than in A. royreba. 3. A. cuzbertsoni revealed strong cytotoxicity for the target CHO cells, whereas A. royreba did not show any specific cytotoxicity. About 80% of the target cells mixed with A. culbertsoni were dead 48 hours after cultivation, and more than 95% of the target cells were dead 72 hours after cultivation. 4. Hydrolase activities in A. culbertsoni cultured with the target cell line were assayed according to the culture time. The activities of acid phosphatase, ${\beta}-N-acetyl$ galactosaminidase, ${\beta}-N-acetyl$ glucosaminidase, ${\alpha}-mannosidase$ and acid proteinase in this pathogenic amoeba were detected higher in amoeba extracts than in culture media up to 120 hours after cultivation, but after 120 hours of cultivation those activities were detected higher in culture media than in the amoeba Iysates. Neutral proteinase activity in A. culbertsoni increased more in EBSS medium than in the Iysate specimens although the activity in the extracts was generally steady according to the cultivation time. Summarizing the above results, it is concluded that there were differences in hydrolase activities between Pathogenic A. culbertsoni and non-pathogenic A. royreba, and that some hydrolase activities were detected remarkably higher in A. culbertsoni which revealed strong cytotoxicity to the target CHO cell line.

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Seed-borne Pathogenic Bacterium Interact with Air-borne Plant Pathogenic Fungus in Rice Fields

  • Jung, Boknam;Park, Jungwook;Kim, Namgyu;Li, Taiying;Kim, Soyeon;Bartley, Laura E.;Kim, Jinnyun;Kim, Inyoung;Kang, Yoonhee;Yun, Ki-Hoon;Choi, Younghae;Lee, Hyun-Hee;Lee, Kwang Sik;Kim, Bo Yeon;Shon, Jong Cheol;Kim, Won Cheol;Liu, Kwang-Hyeon;Yoon, Dahye;Kim, Suhkman;Ji, Sungyeon;Seo, Young Su;Lee, Jungkwan
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.33-33
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    • 2018
  • Air-borne plant pathogenic fungus Fusarium graminearum and seed-borne plant pathogenic bacterium Burkholderia glumae are cause similar disease symptoms in rice heads. Here we showed that two pathogens frequently co-isolated in rice heads and F. graminearum is resistant to toxoflavin produced by B. glumae while other fungal genera are sensitive to the toxin. We have tried to clarify the resistant mechanism of F. graminearum against toxoflavin and the ecological reason of co-existence of the two pathogens in rice. We found that F. graminearum carries resistance to toxoflavin as accumulating lipid in fungal cells. Co-cultivation of two pathogens resulted in increased conidia and enhanced chemical attraction and attachment of the bacterial cells to the fungal conidia. Bacteria physically attached to fungal conidia, which protected bacterium cells from UV light and allowed disease dispersal. Chemotaxis analysis showed that bacterial cells moved toward the fungal exudation compared to a control. Even enhanced the production of phytotoxic trichothecene by the fungal under presence of toxoflavin and disease severity on rice heads was significantly increased by co-inoculation rather than single inoculation. This study suggested that the undisclosed potentiality of air-born infection of bacteria using the fungal spores for survival and dispersal.

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A Black Mold of Pepper Fruites Caused by Alternaria alternata (Alternaria alternata에 의한 고추 열매의 검은곰팡이병)

  • Kim Wan Gyu;Yu Seung Hun
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.67-71
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    • 1985
  • A severe infection on pepper fruits by a black mold fungus was observed in the fields, Suweon. Conidia of Alternaria sp. were isolated from the infected fruits. Based on the morphological characters and size of the conidia, the fungus was identified as Alrernaria alternata (Fries) Keissler. The single spore isolates of A. alternata were weakly or not pathogenic on non-wounded pepper fruits, but highly virulent on fruits wounded with sandpaper or heated-glass rod. The infection on the wounded fruits were more severe on red fruits than on immature green ones. No visible infection was found on leaves and stems when 60-day-old plants of pepper were inoculated with the fungus. In July of 1984, the field losses of pepper fruits caused by black mold were $2.0\~22.5\%$ in Suweon.

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A Nematode Infection in the Epithelial Tissue of Cultured Rockfish Sebastes schlegeli in Cheonsu Bay, Western Korea (서해안 천수만 일대 양식 조피볼락(Sebastes schlegeli)의 상피 선충 감염현황)

  • Seo, Han-Gill;Seo, Jung Soo;Ryu, Min-Kyung;Lee, Eun Hye;Kwon, Se Ryun;Kang, Jong Soon;No, Yun-San;Choi, Hye-Sung;Jung, Sung Hee;Han, Hyun-Ja
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.5
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    • pp.603-610
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    • 2014
  • We investigated a nematode infection in the epithelial tissue of rockfish Sebastes schlegeli cultured in Cheonsu Bay, western Korea, from May 2013 to April 2014. Nematodes infected the epithelial tissues of various external organs, including the fins, operculum, nares, mouth, and head. Over a 1-year period, the overall nematode infection rate in rockfish was 55% (n=89/163): 56.7% (n=17/30) in May 2013, 88% (n=29/33) in August 2013, 0% (n=0/30) in October 2013, 70% (n=21/30) in January 2014, and 55% (n=22/40) in April 2014. During this study, the only mass mortality of cultured rockfish in Cheonsu Bay was in August 2013, when we identified skin wounds on the rockfish caused by the nematodes escaping from their hosts. During this period, the accumulated mortality for 2 weeks was 1.4-22.4% in different farms. In addition, several pathogenic bacteria (Photobacterium damselae, Vibrio spp., and Streptococcus iniae) were isolated from the moribund rockfish; these were thought to be transmitted through the skin wounds caused by the nematodes.

Different Clinical Outcomes of Entamoeba histolytica in Malaysia: Does Genetic Diversity Exist?

  • Anuar, Tengku Shahrul;Al-Mekhlafi, Hesham M.;Abdul Ghani, Mohamed Kamel;Azreen, Siti Nor;Salleh, Fatmah Md.;Ghazali, Nuraffini;Bernadus, Mekadina;Moktar, Norhayati
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.231-236
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    • 2013
  • The present study was conducted to investigate the clinical outcomes of Entamoeba histolytica infection in symptomatic and asymptomatic Orang Asli (aborigine) communities in Malaysia. Examination was performed on 500 stool samples obtained from Orang Asli communities in 3 different states using formalin-ether concentration, trichrome staining, and single-round PCR techniques. Out of 500 stool samples, single infection of E. histolytica, Entamoeba dispar, and Entamoeba moshkovskii was identified in 3.2%, 13.4%, and 1%, respectively. In addition, 10 samples had mixed infections with E. histolytica and E. dispar. Six samples containing E. dispar were also positive for E. moshkovskii, and only 2 samples had E. histolytica in association with E. dispar and E. moshkovskii. Seventeen E. histolytica-positive samples were from symptomatic subjects, whereas the remaining 11 samples came from asymptomatic subjects. These findings suggest a predominant distribution of pathogenic potential of E. histolytica strains in this community. Therefore, further studies on genotyping of E. histolytica is required, to find out association between E. histolytica genotype and the outcome of the infection.

Microbiological Safety of Infant Formula and Baby Food (조제분유와 이유식의 미생물학적 안전성)

  • Lee, Seung-Bae;Choe, Jae-Won;Choe, Seok-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.23 no.1
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    • pp.65-71
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    • 2005
  • Powered infant formula and baby food contaminated with Enterobacter sakazakii were reported to cause infection among infants and to be associated with sporadic cases and outbreaks of sepsis, menigitis, cerebritis, and necrotizing enterocolitis. Salmonella contamination of infant formula has also been responsible for multiple outbreaks. Other species of Enterobacteriaceae in powdered infant formula may be causative agents, about which there has been no report. Other pathogenic bacteria have been isolated from powdered infant formula but they were not associated with outbreaks among infant. While Enterobacter sakazakii caused disease in all age groups, premature infants under 28 days old and with birth weight are most sensitive to its infection. Even if low contamination level of the bacteria in powdered infant formula and baby food may not cause infection, the possibility to multiplicate during preparation and storage of reconstituted formula may increase. The etiological factors and pathogenecity of S. sakazakii have not been elucidated. There were wide variability in phenotype and genotype between its strains. S. sakazakii has been isolated from factory facility and surroundings more frequently than Salmonella and thus factory environment should be the source for post-processing contamination of the formula with S. sakazakii. Considering current technology to manufacture power infant formula and baby food it is impossible to sterilize powdered formula but the frequency of outbreak hazard by S. sakazakii can be reduced by pasteurizing the formula base before drying and shortening storage time of the reconstituted formula.

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Study on the Status on Intestinal Protozoan Infections in the children of Specific Groups (특수집단아동(特殊集團兒童)에 있어서의 원충류감염(原虫類感染) 실태조사(實態調査))

  • Choung, Ji-Tae;Uh, Kwang-Bon;Lee, Joon-Sang
    • Journal of agricultural medicine and community health
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    • v.6 no.1
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    • pp.68-72
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    • 1981
  • This study was undertaken to evaluate the status of intestinal protozoan infections in the children of specific groups. A total number of 441(male 250; female 191) consisting of 224 children of national feeble minded children institute, 59 Han-Bit blindness students, 158 children of local mental retardation children institute(Kang-Dong Ku) of specific groups were examined. The method employed was formalin-ether sedimentation technique with Lugol's iodine stain. The results were as follows: 1) The positive rates for any kinds of protozoan cysts were 13.8%(72 positives out of 441 samples) with sexual difference of 14.0% in male and 12.6% in female. 2) In the institute differences of positive rates of protozoan cysts were 15.2% in national feeble minded children institute, 16.9% in Han-Bit blindness school, and local mental retardation children institute(Kang-Dong Ku). 3) Cysts of Entamoeba coli were recovered in 8.6% and cysts of pathogenic amoeba, Entamoeba histolytica were found in 1.4%. Cysts of Endolimax nana (3.4%). Giardia lamblia (2.9%) were found in this study. 4) The degree of multiple infections of protozoan cysts were; single infection 81.4%, double infection 16.9% and quadriple infection 1.7% respectively.

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Reovirus and Tumor Oncolysis

  • Kim, Man-Bok;Chung, Young-Hwa;Johnston, Randal N.
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.187-192
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    • 2007
  • REOviruses (Respiratory Enteric Orphan viruses) are ubiquitous, non-enveloped viruses containing 10 segments of double-stranded RNA (dsRNA) as their genome. They are common isolates of the respiratory and gastrointestinal tract of humans but are not associated with severe disease and are therefore considered relatively benign. An intriguing characteristic of reovirus is its innate oncolytic potential, which is linked to the transformed state of the cell. When immortalized cells are transfected in vitro with activated oncogenes such as Ras, Sos, v-erbB, or c-myc, they became susceptible to reovirus infection and subsequent cellular lysis, indicating that oncogene signaling pathways are exploited by reovirus. This observation has led to the use of the virus in clinical trials as an anti-cancer agent against oncogenic tumors. In addition to the exploitation of oncogene signaling, reovirus may further utilize host immune responses to enhance its antitumor activity in vivo due to its innate interferon induction ability. Reovirus is, however, not entirely benign to immunocompromised animal models. Reovirus causes so-called "black feet syndrome" in immunodeficient mice and can also harm neonatal animals. Because cancer patients often undergo immunosuppression due to heavy chemo/radiation-treatments or advanced tumor progression, this pathogenic response may be a hurdle in virus-based anticancer therapies. However, a genetically attenuated reovirus variant derived from persistent reovirus infection of cells in vitro is able to exert potent anti-tumor activity with significantly reduced viral pathogenesis in immunocompromised animals. Importantly, in this instance the attenuated, reovirus maintains its oncolytic potential while significantly reducing viral pathogenesis in vivo.

Draft genome sequence of lytic bacteriophage KP1 infecting bacterial pathogen Klebsiella pneumoniae (병원균 Klebsiella pneumoniae를 감염시키는 용균 박테리오파지 KP1의 유전체 염기서열 초안)

  • Kim, Youngju;Bang, Ina;Yeon, Young Eun;Park, Joon Young;Han, Beom Ku;Kim, Hyunil;Kim, Donghyuk
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.152-154
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    • 2018
  • Klebsiella pneumoniae is a Gram-negative, rod-shape bacterium causing disease in human and animal lungs. K. pneumoniae has been often found to gain antimicrobial resistance, thus it has been difficult to treat K. pneumoniae infection with antibiotics. For such infection, bacteriophage can provide an alternative approach for pathogenic bacterial infection with antimicrobial resistance, because of its sensitivity and specificity to the host bacteria. Bacteriophage KP1 was isolated in sewage and showed specific infectivity to K. pneumoniae. Here, we report the draft genome sequence of Klebsiella pneumoniae phage KP1. The draft genome of KP1 is 167,989 bp long, and the G + C content is 39.6%. The genome has 295 predicted ORFs and 14 tRNA genes. In addition, it encodes various enzymes which involve in lysis of the host cell such as lysozyme and holin.