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Abstract: Background Cancer genome and transcriptome analyses advanced our understanding of cancer biology. We performed transcriptome analysis of all known genes of peptidases also called proteases and their endogenous inhibitors in glioblastoma multiforme (GBM), which is one of the most aggressive and deadly types of brain cancers, where unbalanced proteolysis is associated with tumor progression. Methods Comparisons were performed between the transcriptomics of primary GBM tumors and unmatched non-malignant brain tissue, and between GBM cell lines (U87-MG and U373) and a control human astrocyte cell line (NHA). Publicly-available data sets and our own datasets were integrated and normalized using bioinformatics tools to reveal protease and protease inhibitor genes with deregulated expression in both malignant versus non-malignant tissues and cells. Results Of the 311 protease genes identified to be differentially expressed in both GBM tissues and cells, 5 genes were highly overexpressed, 2 genes coding for non-peptidase homologues transferrin receptor (TFRC) and G protein-coupled receptor 56 (GPR56), as well as 3 genes coding for the proteases endoplasmic reticulum aminopeptidase 2 (ERAP2), glutamine-fructose-6-phosphate transaminase 2 (GFPT2) and cathepsin K (CTSK), whereas one gene, that of the serine protease carboxypeptidase E (CPE) was strongly reduced in expression. Seventy five protease inhibitor genes were differentially expressed, of which 3 genes were highly overexpressed, the genes coding for stefin B (CSTB), peptidase inhibitor 3 (PI3 also named elafin) and CD74. Seven out of 8 genes (except CSTB) were validated using RT-qPCR in GBM cell lines. CTSK overexpression was validated using RT-qPCR in GBM tissues as well. Cathepsin K immunohistochemical staining and western blotting showed that only proteolytically inactive proforms of cathepsin K were overexpressed in GBM tissues and cells. Conclusions The presence of high levels of inactive proforms of cathepsin K in GBM tissues and cells indicate that in GBM the proteolytic/collagenolytic role is not its primary function but it plays rather a different yet unknown role.
Keywords: glioblastoma multiforme, genes
Published in DiRROS: 02.08.2024; Views: 374; Downloads: 216
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Abstract: In the field, plants are challenged by more than one biotic stressor at the same time. In this study, the molecular interactions between potato (Solanum tuberosum L.), Colorado potato beetle (Leptinotarsa decemlineata Say; CPB) and Potato virus YNTN (PVYNTN) were investigated through analyses of gene expression in the potato leaves and the gut of the CPB larvae, and of the release of potato volatile compounds. CPB larval growth was enhanced when reared on secondary PVYNTN-infected plants, which was linked to decreased accumulation of transcripts associated with the antinutritional properties of potato. In PVYNTN-infected plants, ethylene signalling pathway induction and induction of auxin response transcription factors were attenuated, while no differences were observed in jasmonic acid (JA) signalling pathway. Similarly to rearing on virus-infected plants, CPB larvae gained more weight when reared on plants silenced in JA receptor gene (coi1). Although herbivore-induced defence mechanism is regulated predominantly by JA, response in coi1-silenced plants only partially corresponded to the one observed in PVYNTN-infected plants, confirming the role of other plant hormones in modulating this response. The release of β-barbatene and benzyl alcohol was different in healthy and PVYNTN-infected plants before CPB larvae infestation, implicating the importance of PVYNTN infection in plant communication with its environment. This was reflected in gene expression profiles of neighbouring plants showing different degree of defence response. This study thus contributes to our understanding of plant responses in agro-ecosystems.
Keywords: insect midgut transcriptional response, gene expression, plant defence, volatile organic compounds, potato
Published in DiRROS: 02.08.2024; Views: 312; Downloads: 181
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Abstract: The ability to identify and locate conspecifics depends on reliable transfer of information between emitter and receiver. For a majority of plant-dwelling insects communicating with substrate-borne vibrations, localization of a potential partner may be a difficult task due to their small body size and complex transmission properties of plants. In the present study, we used the leafhopper Scaphoideus titanus as a model to investigate duetting and mate searching associated with pair formation. Studying these insects on a natural substrate, we showed that the spatio-temporal structure of a vibrational duet and the perceived intensity of partner's signals influence the mating behaviour. Identification, localization and courtship stages were each characterized by a specific duet structure. In particular, the duet structure differed in synchronization between male and female pulses, which enables identification of the partner, while the switch between behavioural stages was associated with the male-perceived intensity of vibrational signals. This suggests that males obtain the information about their distance from the female and optimize their strategy accordingly. More broadly, our results show that even in insects smaller than 1 cm, vibrational signals provide reliable information needed to find a mating partner.
Keywords: mating behaviour, identification, localization, courtship, vibrational communication
Published in DiRROS: 02.08.2024; Views: 330; Downloads: 181
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Abstract: The risk posed by complex chemical mixtures in the environment to wildlife and humans is increasingly debated, but has been rarely tested under environmentally relevant scenarios. To address this issue, two mixtures of 14 or 19 substances of concern (pesticides, pharmaceuticals, heavy metals, polyaromatic hydrocarbons, a surfactant, and a plasticizer), each present at its safety limit concentration imposed by the European legislation, were prepared and tested for their toxic effects. The effects of the mixtures were assessed in 35 bioassays, based on 11 organisms representing different trophic levels. A consortium of 16 laboratories was involved in performing the bioassays. The mixtures elicited quantifiable toxic effects on some of the test systems employed, including i) changes in marine microbial composition, ii) microalgae toxicity, iii) immobilization in the crustacean Daphnia magna, iv) fish embryo toxicity, v) impaired frog embryo development, and vi) increased expression on oxidative stress-linked reporter genes. Estrogenic activity close to regulatory safety limit concentrations was uncovered by receptor-binding assays. The results highlight the need of precautionary actions on the assessment of chemical mixtures even in cases where individual toxicants are present at seemingly harmless concentrations.
Keywords: morje, analizna kemija, biologija, evtrofikacija, hranilne snovi, fitoplankton, onesnaževanje, Piranski zaliv
Published in DiRROS: 02.08.2024; Views: 346; Downloads: 265
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Abstract: Long-term experimental studies suggest that, under transient anoxic conditions, redox fronts within the sediment shift upwards, causing sequential rise and fall of benthic fluxes of reduced species (Mn(II), Fe(II) and S(-II)). Infaunal benthic organisms are associated with different redox fronts as micro-habitats and must be affected by such changes during natural hypoxia events. In order to document the geochemical evolution of the sediment during prolonged anoxia in the framework of an in situ experiment designed to mimic natural conditions, benthic chambers were deployed on the seafloor of the Northern Adriatic and sampled after 9, 30 and 315 days of incubation. Oxygen and sulfide were measured continuously in the early stages (9 days) of the experiment. High-resolution pore water profiles were sampled by DET probes and redox-sensitive species (S(VI), Mn(II) and Fe(II)) and alkalinity were measured. Starting oxygen saturation was about 80% within the chamber. After 7 days, anoxia was established in the bottom waters within the chambers. Mn(II) and Fe(II) started diffusing towards the anoxic water column until they reached the surficial sediment. Being reoxidized there, Mn and Fe reprecipitated, giving a rusty coloration to the seafloor. Infaunal species appeared at the sediment surface. After 20 days, all macro-organisms were dead. Decomposition of macro-organisms at the sediment–water interface generated S(-II) within the entire height of the chamber, leading to a downward flux of sulfides into the sediment, where they were quickly oxidized by metallic oxides or precipitated as FeS. S(-II) was below detection in the water column and pore waters at the end of the experiment. Our results suggest that S(-II) enrichment in the water column of coastal systems, which are episodically anoxic, is strongly controlled by the biomass of benthic macrofauna and its decay during anoxia, whereas its residence time in the water column is controlled by iron availability (as solid oxides or as dissolved reduced cations) within the sediment, even without water circulation.
Keywords: Adriatic Sea, marine sediments, anoxia, Northern Adriatic
Published in DiRROS: 02.08.2024; Views: 305; Downloads: 234
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