- Publikační typ
- abstrakt z konference MeSH
Many lepidopteran larvae produce silk secretions to build feeding tubes and cocoons that play important protective roles in their lives. Recent research on the silk of bombycoid and pyralid moths has shown that it contains several highly abundant silk components with remarkable mechanical properties. It was also found to contain a number of other proteins of which the functions have yet to be identified. To gain an overview of the silk composition in more primitive lepidopteran species and to identify the core silk components common to most species, we analyzed the cocoon proteins of Tineola bisselliella, which belongs to the basal ditrysian moth line. Using de novo transcriptome sequencing combined with mass spectrometry (MS)-based proteomics, we detected more than 100 secretory proteins in the silk cocoons. Fibroin, sericins, and protease inhibitors were found to be the most abundant proteins, along with several novel candidate silk components. We also verified the tissue and developmental stage specificity of the silk protein expression and characterized the morphology of both the silk glands and silk in T. bisselliella. Our study provides a detailed analysis of silk in the primitive moth, expands the known set of silk-specific genes in Lepidoptera, and helps to elucidate their evolutionary relationships.
- MeSH
- biologická evoluce * MeSH
- fibroiny metabolismus MeSH
- hedvábí * chemie genetika metabolismus MeSH
- hmyzí proteiny genetika metabolismus MeSH
- inhibitory proteas metabolismus MeSH
- larva genetika metabolismus fyziologie MeSH
- můry * genetika metabolismus fyziologie MeSH
- proteomika metody MeSH
- sericiny metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb), each year causing millions of deaths. In this article, we present the synthesis and biological evaluations of new potential antimycobacterial compounds containing a fragment of the first-line antitubercular drug pyrazinamide (PZA), coupled with methyl or ethyl esters of selected amino acids. The antimicrobial activity was evaluated on a variety of (myco)bacterial strains, including Mtb H37Ra, M. smegmatis, M. aurum, Staphylococcus aureus, Pseudomonas aeruginosa, and fungal strains, including Candida albicans and Aspergillus flavus. Emphasis was placed on the comparison of enantiomer activities. None of the synthesized compounds showed any significant activity against fungal strains, and their antibacterial activities were also low, the best minimum inhibitory concentration (MIC) value was 31.25 μM. However, several compounds presented high activity against Mtb. Overall, higher activity was seen in derivatives containing ʟ-amino acids. Similarly, the activity seems tied to the more lipophilic compounds. The most active derivative contained phenylglycine moiety (PC-ᴅ/ʟ-Pgl-Me, MIC < 1.95 μg/mL). All active compounds possessed low cytotoxicity and good selectivity towards Mtb. To the best of our knowledge, this is the first study comparing the activities of the ᴅ- and ʟ-amino acid derivatives of pyrazinamide as potential antimycobacterial compounds.
- MeSH
- aminokyseliny chemie farmakologie MeSH
- antibakteriální látky farmakologie MeSH
- antituberkulotika farmakologie MeSH
- Aspergillus flavus účinky léků MeSH
- buňky Hep G2 MeSH
- Candida albicans účinky léků MeSH
- chromatografie kapalinová MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium smegmatis účinky léků MeSH
- Mycobacterium tuberculosis účinky léků MeSH
- optická otáčivost MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- Pseudomonas aeruginosa účinky léků MeSH
- pyrazinamid chemie farmakologie MeSH
- Staphylococcus aureus účinky léků MeSH
- tuberkulóza farmakoterapie MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Understanding the tradeoffs that result from successful infection responses is central to understanding how life histories evolve. Gaining such insights, however, can be challenging, as they may be pathogen specific and confounded with experimental design. Here, we investigated whether infection from gram positive or negative bacteria results in different physiological tradeoffs, and whether these infections impact life history later in life (post-diapause development), in the butterfly Pieris napi. During the first 24 h after infection (3, 6, 12, and 24 h), after removing effects due to injection, larvae infected with Micrococcus luteus showed a strong suppression of all non-immunity related processes while several types of immune responses were upregulated. In contrast, this tradeoff between homeostasis and immune response was much less pronounced in Escherichia coli infections. These differences were also visible long after infection, via weight loss and slower development, as well as an increased mortality at higher infection levels during later stages of development. Individuals infected with M. luteus, compared to E. coli, had a higher mortality rate, and a lower pupal weight, developmental rate and adult weight. Further, males exhibited a more negative impact of infection than females. Thus, immune responses come at a cost even when the initial infection has been overcome, and these costs are likely to affect later life history parameters with fitness consequences.
- Publikační typ
- časopisecké články MeSH
Lepidopteran silk is a complex assembly of proteins produced by a pair of highly specialized labial glands called silk glands. Silk composition has been examined only in a handful of species. Here we report on the analysis of silk gland-specific transcriptomes from three developmental stages of the greater wax moth, Galleria mellonella, combined with proteomics, Edman microsequencing and northern blot analysis. In addition to the genes known earlier, we identified twenty seven candidate cDNAs predicted to encode secretory proteins, which may represent novel silk components. Eight were verified by proteomic analysis or microsequencing, and several others were confirmed by similarity with known silk genes and their expression patterns. Our results revealed that most candidates encode abundant secreted proteins produced by middle silk glands including ten sericins, two seroins, one or more mucins, and several sequences without apparent similarity to known proteins. We did not detect any novel PSG-specific protein, confirming that there are only three fibroin subunits. Our data not only show that the number of sericin genes in the greater wax moth is higher than in other species thus far examined, but also the total content of soluble proteins in silk is twice as high in G. mellonella than in B. mori or A. yamamai. Our data will serve as a foundation for future identification and evolutionary analysis of silk proteins in the Lepidoptera.
- MeSH
- fylogeneze MeSH
- glykoproteiny chemie genetika metabolismus MeSH
- hedvábí genetika metabolismus MeSH
- hmyzí proteiny chemie genetika metabolismus MeSH
- larva genetika růst a vývoj metabolismus MeSH
- muciny chemie genetika metabolismus MeSH
- můry genetika růst a vývoj metabolismus MeSH
- proteom * MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- sericiny chemie genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The pathogenic effect of mutant HTT (mHTT) which causes Huntington disease (HD) are not restricted to nervous system. Such phenotypes include aberrant immune responses observed in the HD models. However, it is still unclear how this immune dysregulation influences the innate immune response against pathogenic infection. In the present study, we used transgenic Drosophila melanogaster expressing mutant HTT protein (mHTT) with hemocyte-specific drivers and examined the immune responses and hemocyte function. We found that mHTT expression in the hemocytes did not affect fly viability, but the numbers of circulating hemocytes were significantly decreased. Consequently, we observed that the expression of mHTT in the hemocytes compromised the immune responses including clot formation and encapsulation which lead to the increased susceptibility to entomopathogenic nematode and parasitoid wasp infections. In addition, mHTT expression in Drosophila macrophage-like S2 cells in vitro reduced ATP levels, phagocytic activity and the induction of antimicrobial peptides. Further effects observed in mHTT-expressing cells included the altered production of cytokines and activation of JAK/STAT signaling. The present study shows that the expression of mHTT in Drosophila hemocytes causes deficient cellular and humoral immune responses against invading pathogens. Our findings provide the insight into the pathogenic effects of mHTT in the immune cells.
- MeSH
- buněčné linie MeSH
- Drosophila melanogaster MeSH
- exprese genu * MeSH
- geneticky modifikovaná zvířata MeSH
- hemocyty imunologie MeSH
- humorální imunita * MeSH
- lidé MeSH
- protein huntingtin genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- lidé MeSH
- osobní vzpomínky jako téma MeSH
- psychoanalýza metody MeSH
- psychoterapie metody výchova MeSH
- terapeutická aliance MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- rozhovory MeSH
Seroins are small lepidopteran silk proteins known to possess antimicrobial activities. Several seroin paralogs and isoforms were identified in studied lepidopteran species and their classification required detailed phylogenetic analysis based on complete and verified cDNA sequences. We sequenced silk gland-specific cDNA libraries from ten species and identified 52 novel seroin cDNAs. The results of this targeted research, combined with data retrieved from available databases, form a dataset representing the major clades of Lepidoptera. The analysis of deduced seroin proteins distinguished three seroin classes (sn1-sn3), which are composed of modules: A (includes the signal peptide), B (rich in charged amino acids) and C (highly variable linker containing proline). The similarities within and between the classes were 31-50% and 22.5-25%, respectively. All species express one, and in exceptional cases two, genes per class, and alternative splicing further enhances seroin diversity. Seroins occur in long versions with the full set of modules (AB1C1B2C2B3) and/or in short versions that lack parts or the entire B and C modules. The classes and the modular structure of seroins probably evolved prior to the split between Trichoptera and Lepidoptera. The diversity of seroins is reflected in proposed nomenclature.
