Polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), together with polycyclic aromatic hydrocarbons (PAHs), represent highly toxic and persistent organic environmental pollutants, especially due to their capability for bioaccumulation in fatty tissues. To observe the environmentally relevant effect of these compounds on earthworms, two soils naturally contaminated with PCDD/Fs and PAHs were used in our experiments. We focused on the role of CuZn- and Mn-superoxide dismutases. We assembled a full-length sequences of these molecules from Eisenia andrei earthworm and confirmed their activity. We demonstrated the significant reduction of CuZn-SOD on both mRNA and enzyme activity levels and increased levels of reactive oxygen species in earthworms kept in PCDD/F-polluted soil, which corresponds to the observed histopathologies of the earthworm intestinal wall and adjacent chloragogenous tissue. The results show an important role of CuZn-SOD in earthworm tissue damage caused by PCDD/Fs present in soil. We did not detect any significant changes in the mRNA expression or activity of Mn-SOD in these earthworms. In earthworms maintained in PAH-polluted soil the activity of both CuZn-SOD and Mn-SOD significantly increased. No histopathological changes were detected in these worms, however significant decrease of coelomocyte viability was observed. This reduced viability was most likely independent of oxidative stress.
- MeSH
- látky znečišťující půdu toxicita MeSH
- Oligochaeta anatomie a histologie účinky léků enzymologie MeSH
- polychlorované dibenzodioxiny toxicita MeSH
- polychlorované dibenzofurany toxicita MeSH
- polycyklické aromatické uhlovodíky toxicita MeSH
- reaktivní formy kyslíku metabolismus MeSH
- superoxid dismutáza 1 genetika metabolismus MeSH
- superoxiddismutasa genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Amyloidosis is a group of diseases caused by the accumulation of insoluble protein aggregates in different parts of the body. Repeated subcutaneous injection of insulin hormones in diabetic patients leads to localized amyloidosis that is found to be cytotoxic. Thus, agents that can dissociate these aggregates are critically needed. In the present study, insulin amyloid dissociation was demonstrated by the treatment of an enzyme lumbrokinase (LK) isolated from earthworm. Thioflavin T (ThT) fluorescence, solution turbidity, particle size analysis, FTIR, CD, atomic force microscopy and cell viability assay were employed to support the dissociation of insulin amyloid in vitro. The small animal optical imaging was used to explore the dissociation of amyloid fibrils in vivo using zebrafish model. The activity of LK towards amyloid dissociation was compared with the standard amyloid fibril degrading agent nattokinase (NK). Our results indicated that LK can be a probable fibril degrading agent for the dissociation of amyloids.
- Klíčová slova
- Lumbrokinasa,
- MeSH
- amyloid biosyntéza metabolismus MeSH
- amyloidní plaky metabolismus MeSH
- amyloidóza etiologie patologie MeSH
- dánio pruhované MeSH
- experimenty na zvířatech MeSH
- fluorescenční spektrometrie metody MeSH
- inzulin škodlivé účinky terapeutické užití MeSH
- Oligochaeta enzymologie MeSH
- proteolýza MeSH
- serinové endopeptidasy farmakologie izolace a purifikace MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Earthworms Eisenia andrei, similarly to other invertebrates, rely on innate defense mechanisms based on the capability to recognize and respond to nonself. Here, we show a correlation between the expression of CCF, a crucial pattern-recognition receptor, and lysozyme, with enzyme activities in the gut of E. andrei earthworms following a microbial challenge. These data suggest that enzyme activities important for the release and recognition of molecular patterns by pattern-recognition molecules, as well as enzymes involved in effector pathways, are modulated during the microbial challenge. In particular, protease, laminarinase, and glucosaminidase activities were increased in parallel to up-regulated CCF and lysozyme expression.
- MeSH
- Bacillus subtilis imunologie MeSH
- celulasy metabolismus MeSH
- Escherichia coli imunologie MeSH
- hexosaminidasy metabolismus MeSH
- messenger RNA metabolismus MeSH
- muramidasa metabolismus fyziologie MeSH
- Oligochaeta enzymologie imunologie mikrobiologie MeSH
- přirozená imunita MeSH
- proteasy metabolismus MeSH
- Saccharomyces cerevisiae imunologie MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Lysozyme is a widely distributed antimicrobial protein having specificity for cleaving the beta-(1,4)-glycosidic bond between N-acetylmuramic acid (NAM) and N-acetylglucosamine (GlcNAc) of peptidoglycan of the bacterial cell walls and thus efficiently contributes to protection against infections caused mainly by Gram-positive bacteria. In the present study, we assembled a full-length cDNA of a novel invertebrate-type lysozyme from Eisenia andrei earthworm (EALys) by RT-PCR and RACE system. The primary structure of EALys shares high homology with other invertebrate lysozymes; however the highest, 72% identity, was shown for the destabilase I isolated from medicinal leech. Recombinant EALys expressed in Escherichia coli exhibited the lysozyme and isopeptidase activity. Moreover, real-time PCR revealed increased levels of lysozyme mRNA in coelomocytes of E. andrei after the challenge with both Gram-positive and Gram-negative bacteria.
- MeSH
- Bacillus subtilis imunologie patogenita MeSH
- bakteriální adheze MeSH
- chitinasy metabolismus MeSH
- Echinodermata genetika MeSH
- endopeptidasy metabolismus MeSH
- Escherichia coli genetika imunologie patogenita MeSH
- glukosamin analogy a deriváty imunologie metabolismus MeSH
- grampozitivní bakteriální infekce imunologie MeSH
- hydrolýza MeSH
- infekce vyvolané Escherichia coli imunologie MeSH
- interakce hostitele a patogenu MeSH
- klonování DNA MeSH
- kyseliny muramové imunologie metabolismus MeSH
- lyasy štěpící vazby C-N metabolismus MeSH
- muramidasa genetika imunologie metabolismus MeSH
- Oligochaeta enzymologie genetika imunologie MeSH
- pijavka lékařská genetika MeSH
- sekvenční homologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
