Introduction. The fungal pathogen Aspergillus fumigatus can induce prolonged colonization of the lungs of susceptible patients, resulting in conditions such as allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis.Hypothesis. Analysis of the A. fumigatus secretome released during sub-lethal infection of G. mellonella larvae may give an insight into products released during prolonged human colonisation.Methodology.Galleria mellonella larvae were infected with A. fumigatus, and the metabolism of host carbohydrate and proteins and production of fungal virulence factors were analysed. Label-free qualitative proteomic analysis was performed to identify fungal proteins in larvae at 96 hours post-infection and also to identify changes in the Galleria proteome as a result of infection.Results. Infected larvae demonstrated increasing concentrations of gliotoxin and siderophore and displayed reduced amounts of haemolymph carbohydrate and protein. Fungal proteins (399) were detected by qualitative proteomic analysis in cell-free haemolymph at 96 hours and could be categorized into seven groups, including virulence (n = 25), stress response (n = 34), DNA repair and replication (n = 39), translation (n = 22), metabolism (n = 42), released intracellular (n = 28) and cellular development and cell cycle (n = 53). Analysis of the Gallerial proteome at 96 hours post-infection revealed changes in the abundance of proteins associated with immune function, metabolism, cellular structure, insect development, transcription/translation and detoxification.Conclusion. Characterizing the impact of the fungal secretome on the host may provide an insight into how A. fumigatus damages tissue and suppresses the immune response during long-term pulmonary colonization.
- MeSH
- Aspergillus fumigatus * metabolismus MeSH
- aspergilóza mikrobiologie metabolismus MeSH
- faktory virulence metabolismus MeSH
- fungální proteiny * metabolismus genetika MeSH
- hemolymfa mikrobiologie metabolismus MeSH
- larva * mikrobiologie MeSH
- můry * mikrobiologie MeSH
- proteom analýza MeSH
- proteomika MeSH
- sekretom metabolismus MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The widespread use of multipotent mesenchymal stromal cell-derived secretome (MSC-sec) requires optimal preservation methods. Lyophilization offers benefits like concentrating the secretome, reducing the storage volume, and making storage conditions more flexible. This study evaluated the influence of storage duration and temperature on lyophilized MSC-sec. The conditioned medium from Wharton's jelly MSCs was stored at - 80 °C or lyophilized with or without trehalose. Lyophilized formulations were kept at - 80 °C, - 20 °C, 4 °C, or room temperature (RT) for 3 and 30 months. After storage and reconstitution, the levels of growth factors and cytokines were assessed using multiplex assay. The storage of lyophilized MSC-sec at - 80 °C ensured biomolecule preservation for 3 and 30 months. Following 3 month storage at 4 °C and RT, a notable decrease occurred in BDNF, bNGF, and sVCAM-1 levels. Prolonged 30 month storage at the same temperatures significantly reduced BDNF, bNGF, VEGF-A, IL-6, and sVCAM-1, while storage at - 20 °C decreased BDNF, bNGF, and VEGF- A levels. Trehalose supplementation of MSC-sec improved the outcome during storage at 4 °C and RT. Proper storage conditions were crucial for the preservation of lyophilized MSC-sec composition. Short-term storage at various temperatures maintained over 60% of the studied growth factors and cytokines; long-term preservation was only adequate at -80 °C.
- MeSH
- cytokiny metabolismus MeSH
- kryoprezervace metody MeSH
- kultivační média speciální chemie MeSH
- kultivované buňky MeSH
- lidé MeSH
- lyofilizace * MeSH
- mezenchymální kmenové buňky * metabolismus cytologie MeSH
- sekretom metabolismus MeSH
- teplota MeSH
- trehalosa metabolismus farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The bacterial secretome represents a comprehensive catalog of proteins released extracellularly that have multiple important roles in virulence and intercellular communication. This study aimed to characterize the secretome of an environmental isolate Pseudomonas aeruginosa S-8 by analyzing trypsin-digested culture supernatant proteins using nano-LC-MS/MS tool. Using a combined approach of bioinformatics and mass spectrometry, 1088 proteins in the secretome were analyzed by PREDLIPO, SecretomeP 2.0, SignalP 4.1, and PSORTb tool for their subcellular localization and further categorization of secretome proteins according to signal peptides. Using the gene ontology tool, secretome proteins were categorized into different functional categories. KEGG pathway analysis identified the secreted proteins into different metabolic functional pathways. Moreover, our LC-MS/MS data revealed the secretion of various CAZymes into the extracellular milieu, which suggests its strong biotechnological applications to breakdown complex carbohydrate polymers. The identified immunodominant epitopes from the secretome of P. aeruginosa showed the characteristic of being non-allergenic, highly antigenic, nontoxic, and having a low risk of triggering autoimmune responses, which highlights their potential as successful vaccine targets. Overall, the identification of secreted proteins of P. aeruginosa could be important for both diagnostic purposes and the development of an effective candidate vaccine.
- MeSH
- bakteriální proteiny * genetika metabolismus imunologie MeSH
- chromatografie kapalinová MeSH
- kovy metabolismus MeSH
- proteomika MeSH
- Pseudomonas aeruginosa * imunologie metabolismus genetika MeSH
- sekretom * metabolismus imunologie MeSH
- tandemová hmotnostní spektrometrie * MeSH
- výpočetní biologie * MeSH
- Publikační typ
- časopisecké články MeSH
Our aim in this study was to characterize and investigate the secretome of Paenibacillus sp. S-12 by nanoLC-MS/MS tool-based analysis of trypsin digested culture supernatant proteins. Using a bioinformatics and combined approach of mass spectrometry, we identified 657 proteins in the secretome. Bioinformatic tools such as PREDLIPO, SecretomeP 2.0, SignalP 4.1, and PSORTb were used for the subcellular localization and categorization of secretome on basis of signal peptides. Among the identified proteins, more than 25% of the secretome proteins were associated with virulence proteins including flagellar, adherence, and immune modulators. Gene ontology analysis using Blast2GO tools categorized 60 proteins of the secretome into biological processes, cellular components, and molecular functions. KEGG pathway analysis identified the enzymes or proteins involved in various biosynthesis and degradation pathways. Functional analysis of secretomes reveals a large number of proteins involved in the uptake and exchange of nutrients, colonization, and chemotaxis. A good number of proteins were involved in survival and defense mechanism against oxidative stress, the production of toxins and antimicrobial compounds. The present study is the first report of the in-depth protein profiling of Paenibacillus bacterium. In summary, the current findings of Paenibacillus sp. S-12 secretome provide basic information to understand its survival and the possible pathogenic mechanism.
In nematodes that invade the gastro-intestinal tract of the ruminant, the process of larval exsheathment marks the transition from the free-living to the parasitic stages of these parasites. To investigate the secretome associated with larval exsheathment, a closed in vitro system that effectively reproduces the two basic components of an anaerobic rumen environment (CO2 and 39 °C) was developed to trigger exsheathment in one of the most pathogenic and model gastrointestinal parasitic nematodes, Haemonchus contortus (barber's pole worm). This study reports the use of multimodal untargeted metabolomics and lipidomics methodologies to identify the metabolic signatures and compounds secreted during in vitro larval exsheathment in the H. contortus infective third-stage larva (iL3). A combination of statistical and chemoinformatic analyses using three analytical platforms revealed a panel of metabolites detected post exsheathment and associated with amino acids, purines, as well as select organic compounds. The major lipid classes identified by the non-targeted lipidomics method applied were lysophosphatidylglycerols, diglycerides, fatty acyls, glycerophospholipids, and a triglyceride. The identified metabolites may serve as metabolic signatures to improve tractability of parasitic nematodes for characterizing small molecule host-parasite interactions related to pathogenesis, vaccine and drug design, as well as the discovery of metabolic biomarkers.
- MeSH
- Haemonchus * MeSH
- hlístice * MeSH
- larva MeSH
- přežvýkavci MeSH
- sekretom MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cíl studie: Přehled problematiky mechanismů receptivity endometria při implantaci embrya a možnosti její diagnostiky. Typ studie: Přehledový článek. Název a sídlo pacoviště: Porodnicko-gynekologická klinika, FN a LF UP Olomouc; Ústav molekulární a translační medicíny, LF UP Olomouc. Výsledky: Endometrium je velmi dynamická tkáň procházející cyklickou proliferací, diferenciací a transportem buněk, zvláště pak buněk imunitního systému. Vše se děje v závislosti na změně cirkulujících ovariálních hormonů estradiolu a progesteronu. Remodelování endometria v době implantace embrya je kontrolováno v časoprostoru intenzitou senescence deciduálních buněk a efektivitou jejich imunologické likvidace. Receptivita endometria může být dnes hodnocena jak na bázi transkriptomického profilování biopsie endometria pomocí ERA systému, tak i proteomickou analýzou endometriálního sekretomu nebo cervikálního hlenu pomocí metod diferenční gelové elektroforézy (DIGE) a hmotnostní spektrometrie (MS). Závěr: Vzhledem k nejnovějším poznatkům v oblasti fyziologie a molekulární biologie endometria by bylo zajímavé aplikovat proteomické přístupy v hledání kandidátních biomarkerů ze vzorků získaných pokud možno neinvazivními postupy a aplikovat je v klinické praxi.
Objective: Literature review of endometrial receptivity in embryo implantation and its diagnostic possibilities. Design: Literature review. Setting: Department of Obstetrics and Gynecology, University Hospital, Faculty of Medicine, Palacky University, Olomouc; Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc. Results: Endometrial tissue is very dynamic, undergoing cyclic proliferation, differentiation and cell transportation, especially of immune system cells under the influence of circulating estradiol and progesterone. Endometrial remodelling during embryo implantation is controlled by decidual cells senescence and effectivity of their immunologic destruction. Endometrial receptivity can be assessed by transcriptomic profiling of endometrial biopsy using ERA system or proteomic analysis of either endometrial secretome or cervical mucus by gel electrophoresis (DIGE) or mass spectrometry (MS). Conclusion: With respect to recent discoveries in endometrial physiology and molecular biology, clinical application of proteomic approaches in research of potential biomarkers of endometrial receptivity could be of interest.
- Klíčová slova
- endometriální receptivita, proteomická analýza,
- MeSH
- cervikální hlen MeSH
- endometrium * MeSH
- implantace embrya * MeSH
- lidé MeSH
- sekretom MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Kmenové buňky se stávají účinným nástrojem v léčbě řady defektů a onemocnění. V poslední době se ukazuje, že terapeutický efekt kmenových buněk spočívá nejenom v jejich integraci a diferenciaci do buněk příslušné tkáně, ale především v jejich parakrinní aktivitě, tj. schopnosti sekretovat trofické a růstové faktory, cytokiny a chemokiny, které mají regenerativní a protizánětlivé účinky. Kondiciované médium (KM) obsahující sekreční produkty kmenových buněk lze proto využít v tzv. bezbuněčné terapii, která představuje alternativu k terapii buněčné s výhodou nižších rizik, možnosti alogenního podání a velkoobjemové výroby. Výsledky preklinických studií potvrzují, že terapeutický efekt KM je srovnatelný s přímou aplikací kmenových buněk. Cílem této práce je shrnout výsledky studií využívající KM z různých typů kmenových buněk v regenerativní medicíně a současně vypracovat přehled faktorů, kterými lze modifikovat buněčnou sekreci a složení KM.
Stem cells become an effective tool for treatment of a variety of defects and diseases. Recently, it appears that the therapeutic effect of stem cells lies not only in their integration and differentiation into cells of the tissue, but especially in their paracrine activity, i.e. the ability to secrete trophic and growth factors, cytokines and chemokines that have regenerative and anti-inflammatory effects. Conditioned medium (CM) containing secretory products of stem cells can thus be used in cell-free therapy which represents an alternative to the cell-based therapy, with advantage of lower risks, the possibility of allogenic administration and mass production. Preclinical studies confirm that the therapeutic effect of CM is comparable to the effect of the application of stem cells. The aim of this paper is to summarize the results of studies using CM from different types of stem cells in regenerative medicine and simultaneously develop an overview of the factors that can modify cellular secretion and the composition of CM.
- Klíčová slova
- bezbuněčná terapie,
- MeSH
- buněčná a tkáňová terapie metody MeSH
- buněčné kultury MeSH
- embryonální kmenové buňky MeSH
- hojení ran MeSH
- kmenové buňky MeSH
- kultivační média speciální * MeSH
- kultivované buňky MeSH
- lidé MeSH
- mezenchymální kmenové buňky MeSH
- parakrinní signalizace MeSH
- preklinické hodnocení léčiv MeSH
- sekretom MeSH
- transplantace kmenových buněk * MeSH
- transplantace mezenchymálních kmenových buněk MeSH
- výzkum kmenových buněk MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
