• Title/Summary/Keyword: Novel pathogen

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Computational Identification of Essential Enzymes as Potential Drug Targets in Shigella flexneri Pathogenesis Using Metabolic Pathway Analysis and Epitope Mapping

  • Narad, Priyanka;Himanshu, Himanshu;Bansal, Hina
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.621-629
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    • 2021
  • Shigella flexneri is a facultative intracellular pathogen that causes bacillary dysentery in humans. Infection with S. flexneri can result in more than a million deaths yearly and most of the victims are children in developing countries. Therefore, identifying novel and unique drug targets against this pathogen is instrumental to overcome the problem of drug resistance to the antibiotics given to patients as the current therapy. In this study, a comparative analysis of the metabolic pathways of the host and pathogen was performed to identify this pathogen's essential enzymes for the survival and propose potential drug targets. First, we extracted the metabolic pathways of the host, Homo sapiens, and pathogen, S. flexneri, from the KEGG database. Next, we manually compared the pathways to categorize those that were exclusive to the pathogen. Further, all enzymes for the 26 unique pathways were extracted and submitted to the Geptop tool to identify essential enzymes for further screening in determining the feasibility of the therapeutic targets that were predicted and analyzed using PPI network analysis, subcellular localization, druggability testing, gene ontology and epitope mapping. Using these various criteria, we narrowed it down to prioritize 5 novel drug targets against S. flexneri and one vaccine drug targets against all strains of Shigella. Hence, we suggest the identified enzymes as the best putative drug targets for the effective treatment of S. flexneri.

Current Status of Plasmodiophora brassicae Researches in Korea

  • Kim, Hong Gi;Lim, Yong Pyo
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.29-29
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    • 2015
  • Clubroot disease is caused by the soil-born obligate plant pathogen Plasmodiophora brassicae. This pathogen can infect all cruciferous vegetables and oil crops, including Brassica rapa, B. oleracea, B. napus, and other Brassica species. Clubroot disease is now considered to be a major problem in Chinese cabbage production in China, Korea, and Japan. We collected several hundreds of P. brassicae infected galls from Korea, and isolated the single spore from the collection. For establishment of novel isolation, and mass-propagation methods for singe spore isolates of P. brassicae pathogen, we developed new filtration method using both cellulose nitrate filter and syringe filter. Accurate detection of P. brassicae pathogen in the field was done by using real-time PCR in the potential infested soil. When we tested the different pathogenicity on commercial Chinese cabbage varieties, P. brassicae from collected galls showed various morphological patterns about clubroot symptom on roots. To date, 8 CR loci have been identified in the B. rapa genome using the quantitative trait loci (QTL) mapping approach, with different resistant sources and isolates. We are trying to develop the molecular marker systems for detect all 8 CR resistant genes. Especially for the study on the interaction between pathogens and CR loci which are not well understood until now, genome wide association studies are doing using the sequenced inbred lines of Chinese cabbage to detect the novel CR genes.

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An integrated DNA barcode assay microdevice for rapid, highly sensitive and multiplex pathogen detection at the single-cell level

  • Jung, Jae Hwan;Cho, Min Kyung;Chung, So Yi;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.276-276
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    • 2013
  • Here we report an integrated microdevice consisting of an efficient passive mixer, a magnetic separation chamber, and a capillary electrophoretic microchannel in which DNA barcode assay, target pathogen separation, and barcode DNA capillary electrophoretic analysis were performed sequentially within 30 min for multiplex pathogen detection at the single-cell level. The intestine-shaped serpentine 3D micromixer provides a high mixing rate to generate magnetic particle-pathogenic bacteria-DNA barcode labelled AuNP complexes quantitatively. After magnetic separation and purification of those complexes, the barcode DNA strands were released and analyzed by the microfluidic capillary electrophoresis within 5 min. The size of the barcode DNA strand was controlled depending on the target bacteria (Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium), and the different elution time of the barcode DNA peak in the electropherogram allows us to recognize the target pathogen with ease in the monoplex as well as in the multiplex analysis. In addition, the quantity of the DNA barcode strand (~104) per AuNP is enough to be observed in the laser-induced confocal fluorescence detector, thereby making single-cell analysis possible. This novel integrated microdevice enables us to perform rapid, sensitive, and multiplex pathogen detection with sample-in-answer-out capability to be applied for biosafety testing, environmental screening, and clinical trials.

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Novel Approaches to Clubroot Management in Western Canada

  • Hwang, Sheau-Fang;Strelkov, Stephen E.
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.49-49
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    • 2015
  • Over the past decade, clubroot has emerged as a major constraint to canola (Brassica napus) production in central Alberta, Canada. The number of fields with confirmed P. brassicae infestations in Alberta has increased steadily from 12 in 2003 to nearly 2,000 in 2014. Management of clubroot on canola has focused on sanitization of field equipment, soil amendments to reduce viable pathogen populations, long rotations out of susceptible crops and cropping of resistant cultivars. Clubroot resistance is the most effective and economical method of disease mitigation, but the recent identification of isolated P. brassicae populations with novel virulence phenotypes capable of overcoming resistance in most canola cultivars highlights the variable nature and adaptability of the pathogen. Recent studies have shown slight reductions in pathogen populations through crop rotations, but much more substantial reductions in spore populations in heavily infested areas near field entrances using fumigants such as Vapam (metam-sodium) or Basamid (dazomet). Greenhouse trials showed that seedling emergence, plant height and root weight increased, while primary and secondary infection and disease severity decreased with increased Basamid dosage. However, field trials showed some phytotoxicity. Application of Vapam at rates of 0.4 to $1.6mL\;L^{-1}$ soil resulted in 12-16 fold reductions in clubroot severity and primary and secondary infection. Vapam also was effective in reducing clubroot severity and improving canola seed yield under field conditions. These studies underscore the need for good resistance stewardship and for the integration of multiple products and practices for successful management of clubroot on canola.

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Phylogenetic and Morphological Identification of the Novel Pathogen of Rheum palmatum Leaf Spot in Gansu, China

  • Wang, Yan;Charkowski, Amy O.;Zeng, Cuiyun;Zhu, Tiantian;Wang, Huizhen;Chen, Honggang
    • Mycobiology
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    • v.44 no.2
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    • pp.93-98
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    • 2016
  • A new leaf spot disease was observed on leaves of Rheum palmatum (Chinese rhubarb) in Northwest China (Gansu Province) starting in 2005. A Septoria-like fungus was isolated and completion of Koch's postulates confirmed that the fungus was the casual agent of the leaf spot disease. Morphology and molecular methods were combined to identify the pathogen. The fungus produced conidiomata pycnidia and the conidia were 2~5 septate, $61.2{\sim}134.1{\mu}m$ in length and $3.53{\sim}5.3{\mu}m$ in width, which is much larger than the known Spetoria species that infects Polygonaceae species. Phylogenic analysis of the internal transcribed spacer region confirmed that this Septoria-like fungus is within the Septoria genus but distinct from known Septoria species. Together, these morphological and phylogenetic data support that the R. palmatum infecting Septoria strain is a newly-described plant pathogenic species.

Characterization of Plasmodium berghei Homologues of T-cell Immunomodulatory Protein as a New Potential Candidate for Protecting against Experimental Cerebral Malaria

  • Cui, Ai;Li, Yucen;Zhou, Xia;Wang, Lin;Luo, Enjie
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.101-115
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    • 2019
  • The pathogenesis of cerebral malaria is biologically complex and involves multi-factorial mechanisms such as microvascular congestion, immunopathology by the pro-inflammatory cytokine and endothelial dysfunction. Recent data have suggested that a pleiotropic T-cell immunomodulatory protein (TIP) could effectively mediate inflammatory cytokines of mammalian immune response against acute graft-versus-host disease in animal models. In this study, we identified a conserved homologue of TIP in Plasmodium berghei (PbTIP) as a membrane protein in Plasmodium asexual stage. Compared with PBS control group, the pathology of experimental cerebral malaria (ECM) in rPbTIP intravenous injection (i.v.) group was alleviated by the downregulation of pro-inflammatory responses, and rPbTIP i.v. group elicited an expansion of regulatory T-cell response. Therefore, rPbTIP i.v. group displayed less severe brain pathology and feverish mice in rPbTIP i.v. group died from ECM. This study suggested that PbTIP may be a novel promising target to alleviate the severity of ECM.

Proteomic Analysis of Shigella Virulence Effectors Secreted under Different Conditions

  • Liu, Xingming;Lu, Lilan;Liu, Xinrui;Liu, Xiankai;Pan, Chao;Feng, Erling;Wang, Dongshu;Niu, Chang;Zhu, Li;Wang, Hengliang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.171-178
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    • 2017
  • A series of novel effector molecules secreted by the type three secretion system (T3SS) of Shigella spp. have been reported in recent years. In this study, a proteomic approach was applied to study T3SS effectors systematically. First, proteins secreted by the S. flexneri wild-type strain after Congo Red induction were separated and identified using two-dimensional electrophoresis to display the relative abundance of all kinds of early effectors for the first time. Then, a gene deletion mutant of known virulence repressor (OspD1) and a gene overexpressed mutant of two known virulence activators (MxiE and IpgC) were constructed and analyzed to discover potential late effectors. Furthermore, the supernatant proteins of gene deletion mutants of two known translocators (IpaB and IpaD), which would constantly secrete effectors, were also analyzed. Among all of the secreted proteins identified in our study, IpaH1.4, IpaH_5, and IpaH_7 have not been reported before. These proteomics data of the secreted effectors will be valuable to understand the pathogenesis of S. flexneri.

Oral Pathogens and Their Antibiotics from Marine Organisms: A Systematic Review of New Drugs for Novel Drug Targets

  • Sehyeok Im;Jun Hyuck Lee;Youn-Soo Shim
    • Journal of dental hygiene science
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    • v.24 no.2
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    • pp.84-96
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    • 2024
  • Background: Recent studies have elucidated the quorum-sensing mechanisms, biofilm formation, inter-pathogen interactions, and genes related to oral pathogens. This review aims to explore the recent expansion of drug targets against oral pathogens and summarize the current research on novel antibiotic substances derived from marine organisms that target oral pathogens. Methods: A comprehensive literature review summarized the novel mechanisms pertaining to quorum-sensing signal transmission systems, biofilm formation, and metabolite exchange in oral pathogens. The amino acid sequences of the 16 proteins identified as potential drug targets were systematically classified and compared across various oral microorganisms. Results: Through a literature review, we identified nine studies researching quorum sensing signaling inhibitors targeting oral pathogens. A comparison of the amino acid sequences of 16 potential drug targets in oral microorganisms revealed significant differences between oral pathogens and beneficial oral symbiotic microorganisms. These findings imply that it is possible to design drugs that can bind more selectively to oral pathogens. Conclusion: By summarizing the results of recent research on the signaling mechanisms that cause pathogenicity, new drug targets against oral pathogens were proposed. Additionally, the current status of developing new antibiotics for oral pathogens using recently developed quorum sensing inhibitors and natural products derived from marine organisms was introduced. Consequently, marine natural products can be used to develop drugs targeting new proteins in oral pathogens.

Novel Sources of Resistance to Phytophthora capsici on Pepper (Capsicum sp.) Landraces from Mexico

  • Retes-Manjarrez, Jesus Enrique;Rubio-Aragon, Walter Arturo;Marques-Zequera, Isidro;Cruz-Lachica, Isabel;Garcia-Estrada, Raymundo Saul;Sy, Ousmane
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.600-607
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    • 2020
  • Phytophthora capsici Leonian is a major pathogen of pepper worldwide and few resistance sources to this pathogen have been identified so far. The goals of this study were to identify new sources of resistance against P. capsici in Capsicum landraces and analyze the relationship between the resistance indicator of plant symptoms and some plant phenotype parameters of plant height, stem width, leaf length and leaf width. Thirty-two landraces of pepper were collected from fourteen states in Mexico. From each population, 36 plants were inoculated with 10,000 zoospores of P. capsici under controlled conditions. This experiment was repeated twice. Out of the 32 landraces, six showed high level of resistance, four showed intermediate resistance and five showed low level of resistance when compared with the susceptible control 'Bravo' and the resistant control 'CM334', indicating that these landraces are promising novel sources of resistance to P. capsici. There was no correlation between the symptoms and plant phenotype parameters. However, these parameters were not affected in the group classified as highly resistant, indicating that P. capsici does not affect the growing of these resistant pepper landraces. The other resistant groups were significantly affected in a differently manner regarding their phenotype, indicating that this pathogen reduce their growth in different ways. This study reports novel resistance sources with great potential that could be used in breeding programs to develop new pepper cultivars with durable resistance to P. capsici.