• Title/Summary/Keyword: toxin A

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Detection and characterization of potential virulence determinants in Staphylococcus pseudintermedius and S. schleiferi strains isolated from canine otitis externa in Korea

  • Gi Yong Lee;Soo In Lee;Ji Heon Park;Sun Do Kim;Geun-Bae Kim;Soo-Jin Yang
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.85.1-85.13
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    • 2023
  • Background: A recent increase in the occurrence of canine skin and soft tissue infections, including otitis externa and pyoderma, caused by antimicrobial-resistant Staphylococcus pseudintermedius and S. schleiferi has become a significant public and veterinary health issues. Objective: We investigated the virulence potentials associated with the occurrence of canine otitis externa in S. pseudintermedius and S. schleiferi. Methods: In this study, the prevalence of genes encoding leukocidins, exfoliative toxins, and staphylococcal enterotoxins (SEs) was investigated using previously characterized S. pseudintermedius (n = 26) and S. schleiferi (n = 19) isolates derived from canine otitis externa. Susceptibility to cathelicidins (K9CATH and PMAP-36) and hydrogen peroxide (H2O2) was also examined in both staphylococcal species. Results: A high prevalence of genes encoding leukocidins (lukS/F-I, lukS1/F1-S, and lukS2/F2-S), exfoliative toxins (siet, expB, and sset), and SEs was identified in both S. pseudintermedius and S. schleiferi isolates. Notably, S. pseudintermedius isolates possessed higher number of SE genes, especially newer SE genes, than S. schleiferi isolates harboring egc clusters. Although no significant differences in susceptibility to K9CATH and H2O2 were observed between the two isolate groups, S. pseudintermedius isolates exhibited enhanced resistance to PMAP-36 compared to S. schleiferi isolates. Conclusions: These findings suggest that high a prevalence of various toxin genes together with enhanced resistance to cathelicidins may contribute to the pathogenicity of S. pseudintermedius and S. schleiferi in canine cutaneous infections.

Characteristics of Harmful Cyanobacteria Occurrence and Toxin Residual in Agricultural Reservoirs of Southern Gyeonggi (경기남부권 농업용 저수지의 유해 남세균 발생 및 독소물질 잔류 특성)

  • Kim Minju;Kim Young-Kee
    • Journal of Korean Society on Water Environment
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    • v.40 no.1
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    • pp.1-10
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    • 2024
  • Algal blooms occur seasonally in the eutrophicated rivers or reservoirs, and some harmful cyanobacteria species produce toxic substances, which are directly or indirectly harmful to the ecosystem and terrestrial animals. So, the monitoring and control of harmful cyanobacteria occurrence and toxins residual in the aquasystem are important to preserve the water environment and secure public health. In this study, the four harmful cyanobacteria occurrences and toxic substance concentrations of two agricultural reservoirs in the southern part of Gyeonggi Province were investigated from August to October 2022. Among four harmful cyanobacteria (Microcystis sp., Anabaena sp., Oscillatoria sp., Aphanizomenon sp.), three kinds of cyanobacteria except Oscillatoria sp. were observed, and Microcystis sp. was the dominant cyanobacteria except for Anabaena sp. dominant result of a sample collected on October at reservoir B. The cell density of cyanobacteria was influenced by season and weather due to the length of daytime and concentrations of organic carbon and nitrogen. Three kinds of microcystin and anatoxin-a were quantitatively analyzed as total (in the cell body and water) and extracellular (in water) concentrations. The maximum total concentrations of anatoxin-a, microcystin-LR, microcystin-RR, and microcystin-YR were 0.1291 ㎍/L, 0.2776 ㎍/L, 0.3721 ㎍/L, and 0.0306 ㎍/L, respectively, in reservoir A and 0.3274 ㎍/L, 0.1495 ㎍/L, 0.2037 ㎍/L, and 0.0153 ㎍/L, respectively, in reservoir B.

Modulation of ATP-Induced Activation of the Muscarinic $K^+$ Channel Activity by Protein Kinase C

  • Kim, Yang-Mi;Park, Hong-Ki;Han, Jae-Hee;Park, Choon-Ok;Hong, Seong-Geun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.6
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    • pp.743-752
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    • 1998
  • The atrial acetylcholine-activated $K^+\;(K_{ACh})$ channel is gated by the pertussis toxin-sensitive inhibitory G $(G_K)$ protein. Earlier studies revealed that ATP alone can activate the $K_{ACh}$ channel via transphosphorylation mediated by nucleoside-diphosphate kinase (NDPK) in atrial cells of rabbit and guinea pig. This channel can be activated by various agonists and also modulated its function by phosphorylation. ATP-induced $K_{ACh}$ channel activation (AIKA) was maintained in the presence of the NDPK inhibitor, suggesting the existence of a mechanism other than NDPK-mediated process. Here we hypothesized the phosphorylation process as another mechanism underlying AIKA and was undertaken to examine what kinase is involved in atrial cells isolated from the rat heart. Single application of 1 mM ATP gradually increased the activity of $K_{ACh}$ channels and reached its maximum $40{\sim}50$ sec later following adding ATP. AIKA was not completely reduced but maintained by half even in the presence of NDPK inhibitor. Neither ADP nor a non-hydrolyzable ATP analogue, AMP-PNP can cause AIKA, while a non-specific phosphatase, alkaline phosphatase blocked completely AIKA. PKC antagonists such as sphingosine or tamoxifen, completely blocked AIKA, whereas PKC catalytic domain increased AIKA. Taken together, it is suggested that the PKC-mediated phosphorylation is partly involved in AIKA.

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Effect of Artemisiae Capillaris Herba on Anti-inflammatory Properties in RAW264.7 Cell Line

  • Kang Seong Yeob;Kim Sung Hoon;Kim Su Myung;Namgoong Uk;Kim Dong Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1832-1842
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    • 2004
  • The present study was performed to examine the possible anti-inflammatory effects of a herbal drug ASCH in RAW264.7 cell line. Inflammation was induced by LPS toxin treatment to RAW264.7 macrophage cell line. Increases in cytokine production such as IL-1β, IL-6. and IL-18, COX-2, NOS-Ⅱ (iNOS), and TNF-alpha were observed at mRNA level in the LPS-treated RAW264.7 cells. Measurement of IL-6, nitric oxide and the reactive oxygen species (ROS) showed increased production of these inflammation mediators. Treatment of ASCH effectively decreased IL-1β protein in a dose-dependent manner, and IL-6 and IL-18 were reduced at 100㎍/㎖ of ASCH concentration. NO production was also decreased by ASCH treatment. A slight inhibition for TNF-alpha in terms of protein, but not mRNA level was obtained by 100㎍/㎖ of ASCH treatment. ASCH treatment to normal RAW264.7 cells did not produce any cytotoxicity, indicting that the action of ASCH was selective to inflammatory cells. Thus, the present data suggest that ASCH may act as an important regulator to alleviate the inflammatory symptoms.

Redesign of an Interhelical Loop of the Bacillus thuringiensis Cry4B delta-endotoxin for Proteolytic Cleavage

  • Krittanai, Chartchai;Lungchukiet, Panida;Ruangwetdee, Sarinthip;Tuntitippawan, Tipparut;Panyim, Sakol;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.34 no.2
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    • pp.150-155
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    • 2001
  • The mosquito-larvicidal Cry4B protein from Bacillus thuringiensis subsp. israelensds was expressed in Escherichia coli. Upon activation by trypsin, the 130-kDa protoxin was processed into the 65-kDa active toxin containing two polypeptide fragments of ca. 47 and ca. 20 kDa. These two polypeptides are products of internal cleavages on the exposed loop connecting helices 5 and 6 in the seven-helical bundle domain. PCR-based mutagenesis was employed to introduce an additional cleavage site into the loop connecting helices 3 and 4. A series of amino acid changes were introduced into the targeted loop, resulting in seven mutant protoxins. Upon digestion with trypsin, a group of mutants with arginine introduced into the loop (EPRNQ, EPNRNQ, EPRNP, ESRNP and SSRNP) produced polypeptide products similar to those of the wild type (EPNNQ). When the loop, SSRNP, was expanded by an insertion of either asparagine (NSSRNP) or valine (VSSRNP), an additional cleavage was detected with proteolytic products of 47,12 and 6 kDa. This cleavage was confirmed to be at the introduced arginine residue by N-terminal sequencing. The mosquito larvicidal assay against Aedes aegypti demonstrated a relatively unchanged toxicity for the mutants without cleavage and reduced toxicity for those with an additional cleavage.

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Identification of Toxin Gene and Antibiotic Resistance of Staphylococcus Aureus Isolated from Agricultural Product Cultivation Environments (농산물 생산 환경에서 분리된 Staphylococcus aureus의 항생제 내성 및 독소 유전자 확인)

  • Park, Su-Hee;Kim, Jeong-Sook;Kim, Kyeong-Yeol;Chung, Duck-Hwa;Shim, Won-Bo
    • Journal of Environmental Health Sciences
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    • v.39 no.5
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    • pp.465-473
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    • 2013
  • Objectives: This study was undertaken to analyze Staphylococcus aureus from cultivation environments for agricultural products and to confirm antibiotic resistance and enterotoxin genes for the isolated S. aureus. Methods: A total of 648 samples were collected from apple, peach, ginseng and balloon flower farms. S. aureus was isolated from soil, agricultural water, personal hygiene elements (hands, gloves and clothes) and work utensils (boxes). Results: S. aureus was detected in a total of 25 samples and 72 strains were isolated. The resistance rate of the isolated S. aureus strains was confirmed at 33.3%, with 24 resistant strains among the total of 72. Fourteen different patterns types were found, and three pattern types (NV, OX, VA) were confirmed most frequently. As result of the detection of enterotoxin gene type, four gene types (sea: 1, sed: 4, seg: all isolated S. aureus, sei: all isolated S. aureus) were analyzed among a total of nine types. Conclusions: This study demonstrates that personal hygiene techniques should be properly managed, such as washing and sterilization before or after work, because agricultural contamination by S. aureus frequently developed through improper management.

Crotamine stimulates phagocytic activity by inducing nitric oxide and TNF-α via p38 and NFκ-B signaling in RAW 264.7 macrophages

  • Lee, Kyung Jin;Kim, Yun Kyu;Krupa, Martin;Nguyen, Anh Ngoc;Do, Bich Hang;Chung, Boram;Vu, Thi Thu Trang;Kim, Song Cheol;Choe, Han
    • BMB Reports
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    • v.49 no.3
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    • pp.185-190
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    • 2016
  • Crotamine is a peptide toxin found in the venom of the rattlesnake Crotalus durissus terrificus and has antiproliferative, antimicrobial, and antifungal activities. Herein, we show that crotamine dose-dependently induced macrophage phagocytic and cytostatic activity by the induction of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α). Moreover, the crotamineinduced expression of iNOS and TNF-α is mediated through the phosphorylation of p38 and the NF-κB signaling cascade in macrophages. Notably, pretreatment with SB203580 (a p38-specific inhibitor) or BAY 11-7082 (an NF-κB inhibitor) inhibited crotamine-induced NO production and macrophage phagocytic and cytotoxic activity. Our results show for the first time that crotamine stimulates macrophage phagocytic and cytostatic activity by induction of NO and TNF-α via the p38 and NF-κB signaling pathways and suggest that crotamine may be a useful therapeutic agent for the treatment of inflammatory disease.

Molecular Classification of Commercial Spirulina Strains and Identification of Their Sulfolipid Biosynthesis Genes

  • Kwei, Chee Kuan;Lewis, David;King, Keith;Donohue, William;Neilan, Brett A.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.359-365
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    • 2011
  • Cyanobacterial strains of the genus Spirulina have recently been identified as an excellent source of sulfolipids, some of which possess anti-HIV properties. Thus, to investigate the distribution of sufolipid biosynthesis pathways in Spirulina, a genetic screening/phylogentic study was performed. Five different strains of Spirulina [Spirulina (Jiangmen), Spirulina sp., S. platensis, S. maxima, and Spirulina seawater] sourced from different locations were initially classified via 16S rDNA sequencing, and then screened for the presence of the sulfolipid biosynthesis genes sqdB and sqdX via a PCR. To assess the suitability of these strains for human consumption and safe therapeutic use, the strains were also screened for the presence of genes encoding nonribosomal peptide synthases (NRPSs) and polyketide synthases (PKSs), which are often associated with toxin pathways in cyanobacteria. The results of the 16S rDNA analysis and phylogenetic study indicated that Spirulina sp. is closely related to Halospirulina, whereas the other four Spirulina strains are closely related to Arthrospira. Homologs of sqdB and sqdX were identified in Spirulina (Jiangmen), Spirulina sp., S. platensis, and the Spirulina seawater. None of the Spirulina strains screened in this study tested positive for NRPS or PKS genes, suggesting that these strains do not produce NRP or PK toxins.

Forskolin Changes the Relationship between Cytosolic $Ca^{2+}$ and Contraction in Guinea Pig Ileum

  • Han, Koon-Hee;Cheon, Gap-Jin;Yeon, Dong-Soo;Kwon, Seong-Chun
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.3
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    • pp.189-194
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    • 2009
  • This study was designed to clarify the mechanism of the inhibitory effect of forskolin on contraction, cytosolic $Ca^{2+}$ level $([Ca^{2+}]_i)$, and $Ca^{2+}$ sensitivity in guinea pig ileum. Forskolin (0.1 nM ${\sim}$ 10 ${\mu}M$) inhibited high $K^+$ (25 mM and 40 mM)- or histamine (3 ${\mu}M$)-evoked contractions in a concentration-dependent manner. Histamine-evoked contractions were more sensitive to forskolin than high $K^+$-evoked contractions. Spontaneous changes in $[Ca^{2+}]_i$ and contractions were inhibited by forskolin (1 ${\mu}M$) without changing the resting $[Ca^{2+}]_i$. Forskoln (10 ${\mu}M$ ) inhibited muscle tension more strongly than $[Ca^{2+}]_i$ stimulated by high $K^+$, and thus shifted the $[Ca^{2+}]_i$-tension relationship to the lower-right. In histamine-stimulated contractions, forskolin (1 ${\mu}M$) inhibited both $[Ca^{2+}]_i$ and muscle tension without changing the $[Ca^{2+}]_i$-tension relationship. In ${\alpha}$-toxin-permeabilized tissues, forskolin (10 ${\mu}M$) inhibited the 0.3 ${\mu}M$ $Ca^{2+}$-evoked contractions in the presence of 0.1 mM GTP, but showed no effect on the $Ca^{2+}$-tension relationship. We conclude that forskolin inhibits smooth muscle contractions by the following two mechanisms: a decrease in $Ca^{2+}$ sensitivity of contractile elements in high $K^+$-stimulated muscle and a decrease in $[Ca^{2+}]_i$ in histamine-stimulated muscle.

A Study on the Kinds(種類), Causes(病因) and Mechanisms(病機) of Malaria(瘧疾) (학질(瘧疾)의 종류(種類)와 병인(病因).병기(病機)에 대한 고찰(考察))

  • Kang, Hyo-Jin;Jeong, Chang-Hyung;Jang, Woo-Chang;Lyu, Jeong-Ah;Baik, You-Sang
    • Journal of Korean Medical classics
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    • v.26 no.2
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    • pp.133-174
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    • 2013
  • Objective : Malaria(瘧疾) is a disease that's main symptom is paroxysm - a cyclical occurrence of sudden coldness followed by rigor and then fever. Since the introduction of the cause and mechanism of malaria(瘧疾) in the "Suwen(素問)", including Cold malaria(寒瘧), Warm malaria(溫瘧), Heat malaria(癉瘧) and Wind malaria(風瘧), there has been over 20 different kinds of malaria, each of which are introduced in multiple medical texts. Method : Through comparison between "Suwen(素問)" and other medical texts, the categories, causes and mechanisms of malaria can be analysed and organized to overview the whole feature of it. Results & Conclusion : External pathogens of malaria(瘧疾) are wind(風), cold(寒), summerheat (暑), dampness(濕), miasmic toxin(瘴), pestilence(疫), ghost(鬼). Internal pathogens of malaria(瘧疾) are dietary irregularities(飮食不節), overexertion and fatigue(勞倦), phlegm(痰), seven emotion(七情). Malaria can be categorized into four groups according to the pathological mechanism that leads to paroxysm. They are latency of disease(伏氣), external contraction(外感), internal damage(內傷), and combination of disease(合病). Malaria-Paroxysm(瘧疾發作) occurs when the three following factors collide strongly : defense qi(衛氣), latent qi(伏邪) and external pathogen(新邪). When collision of the three factors takes place in the interior(裏), the body experiences chills. When it takes place in the exterior(表), the body experiences fever. The cyclical occurrence of Malaria-Paroxysm follows the circulation of defense qi.