- MeSH
- krevní obraz * MeSH
- lidé MeSH
- lymfocyty chemie MeSH
- monocyty chemie MeSH
- neutrofily chemie MeSH
- trombocyty chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Platelet concentrates and especially their further product platelet lysate, are widely used as a replacement for cell culturing. Platelets contain a broad spectrum of growth factors and bioactive molecules that affect cellular fate. However, the cellular response to individual components of the human platelet concentrate is still unclear. The aim of this study was to observe cellular behavior according to the individual components of platelet concentrates. The bioactive molecule content was determined. The cells were supplemented with a medium containing 8% (v/v) of platelet proteins in plasma, pure platelet proteins in deionized water, and pure plasma. The results showed a higher concentration of fibrinogen, albumin, insulin growth factor I (IGF-1), keratinocyte growth factor (KGF), and hepatocyte growth factor (HGF), in the groups containing plasma. On the other hand, chemokine RANTES and platelet-derived growth factor bb (PDGF-bb), were higher in the groups containing platelet proteins. The groups containing both plasma and plasma proteins showed the most pronounced proliferation and viability of mesenchymal stem cells and fibroblasts. The platelet proteins alone were not sufficient to provide optimal cell growth and viability. A synergic effect of platelet proteins and plasma was observed. The data indicated the importance of plasma in platelet lysate for cell growth.
- MeSH
- albuminy MeSH
- becaplermin metabolismus MeSH
- buněčné kultury metody MeSH
- chemokiny metabolismus MeSH
- fibrinogen metabolismus MeSH
- fibroblastový růstový faktor 7 MeSH
- fibroblasty metabolismus MeSH
- hepatocytární růstový faktor MeSH
- insulinu podobný růstový faktor I MeSH
- krevní plazma chemie MeSH
- kultivační média chemie MeSH
- lidé MeSH
- mezenchymální kmenové buňky metabolismus MeSH
- plazma bohatá na destičky metabolismus MeSH
- proliferace buněk účinky léků MeSH
- protoonkogenní proteiny c-sis metabolismus MeSH
- trombocyty chemie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Proteomika je stále viac používanou metódou vo viacerých odvetviach medicíny, avšak jej aplikácia v psychiatrii je stále ve3⁄4mi obmed-zená. V našej pilotnej štúdii sme identifikovali alterovanú skupinu proteínov s použitím proteomickej analýzy mozgovomiechovéhomoku a krvných doštièiek u suicidálnych jedincov v rámci metabolických dráh glykolýzy/glukoneogenézy, 14-3-3 sprostredkovanej sig-nalizácie a komplementovej a koagulaènej kaskády. Na základe našich výsledkov predpokladáme, že zmeny metabolizmu glukózy spoluso zmenami komplementovej a koagulaènej kaskády a 14-3-3 sprostredkovanej signalizácie môžu hraś úlohu v neurobiológii samovrážd v zmysle zníženej utilizácie glukózy a zhoršenej odpovede na oxidatívny stres. Skupinu proteínov identifikovaných v našej pilotnej štúdii, tzv. potencionálnych kandidátov biomarkerov suicidality, je však nevyhnutné overiś ïalšími výskumami na väèšom súbore jedincov.
Despite the fact that proteomic analysis is becoming widely used in various medical fields its use in psychiatry is still very limited. We decided to study suicidal behaviour via cerebrospinal fluid and platelets with the use of proteomics. We identified a group of alte-red proteins in suicidal patients in the metabolic pathways of glycolysis/gluconeogenesis, complement and coagulation cascade and14-3-3 mediated signaling pathway. Based on these findings we suppose that alterations of glucose metabolism (especially utilisation of glucose and altered response to oxidative stress) together with alterations in complement and coagulation cascade and 14-3-3 mediated signaling pathway may play a role in the neurobiology of suicide. However, further research is needed to clarify whether the identified group of proteins can be used as a potential peripheral biomarker for suicidal behaviour.
- MeSH
- biologické markery krev MeSH
- glukoneogeneze MeSH
- glykolýza MeSH
- hemokoagulace MeSH
- komplement MeSH
- lidé středního věku MeSH
- lidé MeSH
- pilotní projekty MeSH
- pitva MeSH
- proteiny 14-3-3 MeSH
- proteiny v mozkomíšním moku * MeSH
- proteiny izolace a purifikace klasifikace MeSH
- proteomika * MeSH
- sebevražda * MeSH
- trombocyty * chemie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
Hemostáza je dynamický proces, který začíná u plodu v průběhu intrauterinního vývoje. Důležitou součástí hemostázy jsou krevní destičky. V souborném referátu jsou prezentovány funkční a morfologická specifika neonatálních destiček, historický přínos profesora Otty Hrodka v diagnostice poruch hemostázy u novorozenců a současné možnosti diagnostiky poruch funkce destiček v novorozeneckém období (hodnocení agregace neonatálních destiček, průtoková cytometrie, agregometrie, tromboelastografie a rotační tromboelastometrie).
Hemostasis is a dynamic process that begins in the fetus within intrauterine development. Blood platelets plays an important part of hemostasis. Functional and morphological changes of neonatal platelets are presented in this review article as well the historical contribution of Professor Otto Hrodka in the diagnosis of neonatal hemostasis disorders Futhermore current possibilities in the diagnosis of platelet dysfunction in the neonatal period are expressed (evaluation of neonatal platelet aggregation, flow cytometry, agregometry, thrombelastography and rotational thrombelastometry).
A three-dimensional scaffold of type I collagen and hydroxyapatite enriched with polycaprolactone nanofibers (Coll/HA/PCL), autologous mesenchymal stem cells (MSCs) in osteogenic media, and thrombocyte-rich solution (TRS) was an optimal implant for bone regeneration in vivo in white rabbits. Nanofibers optimized the viscoelastic properties of the Coll/HA scaffold for bone regeneration. MSCs and TRS in the composite scaffold improved bone regeneration. Three types of Coll/HA/PCL scaffold were prepared: an MSC-enriched scaffold, a TRS-enriched scaffold, and a scaffold enriched with both MSCs and TRS. These scaffolds were implanted into femoral condyle defects 6 mm in diameter and 10-mm deep. Untreated defects were used as a control. Macroscopic and histological analyses of the regenerated tissue from all groups were performed 12 weeks after implantation. The highest volume and most uniform distribution of newly formed bone occurred in defects treated with scaffolds enriched with both MSCs and TRS compared with that in defects treated with scaffolds enriched by either component alone. The modulus of elasticity in compressive testing was significantly higher in the Coll/HA/PCL scaffold than those without nanofibers. The composite Coll scaffold functionalized with PCL nanofibers and enriched with MSCs and TRS appears to be a novel treatment for bone defects.
- MeSH
- hydroxyapatit chemie MeSH
- kolagen chemie MeSH
- králíci MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky cytologie metabolismus MeSH
- nanovlákna chemie MeSH
- polyestery chemie MeSH
- regenerace kostí * MeSH
- tkáňové podpůrné struktury chemie MeSH
- trombocyty chemie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Autorka se zaměřila na destičkové faktory z alfa granulí trombocytů, konkrétně PDGF, IGF 1, TGF a VEGF. Trombocyty byly nejprve destruovány termodestrukcí, aby se faktory vyplavily do plazmy. Plazma byla defibrinovaná tepelně, čímž došlo jak k vysrážení koagulačních faktorů, tak k inaktivaci komplementu. Po centrifugaci byl destičkový lyzát přetlačen plazmaextraktorem do ozařovacího vaku. Byl přidán roztok Riboflavinu a destičkový lyzát byl virově a bakteriálně inaktivován systémem Mirasol. Konečný roztok byl lyofilizován. Autorka validovala každý kritický výrobní krok. Destičkové faktory byly měřeny na začátku procesu, po mikrobiální inaktivaci a po lyofilizaci. Výrobní proces neměl významný vliv na hladinu destičkových faktorů. Toto zjištění je zásadní pro další využití destičkového lyzátu.
The author focused on factors from platelet alpha granules, specifically the PDGF, IGF-1, a VEGF and TGF. Platelets were first broken by deep frozen that had washed factors into the plasma. Plasma was heat-treated, thereby leading to precipitation of coagulation factors, as well as to inactivate the complement. After centrifugation platelet lysate was pushed to the bag. Riboflavin solution was added and platelet lysate was viral and bacterial inactivated by Mirasol system. The final solution was freeze-dried. The author of validated each critical step. Platelet factors were measured at the beginning of the process, following the inactivation of microbial and after freeze-dried. The production process had not a significant impact on the level of platelet factors. This finding is crucial for the other using of platelet lysate.
- MeSH
- dárci krve MeSH
- destičkový růstový faktor * MeSH
- imunoanalýza * metody MeSH
- imunochemie MeSH
- insulinu podobný růstový faktor I analýza MeSH
- klinické laboratorní techniky MeSH
- lidé MeSH
- luminiscenční měření metody MeSH
- lyofilizace * MeSH
- plazma bohatá na destičky * MeSH
- separace krevních složek MeSH
- transformující růstové faktory MeSH
- trombocyty * chemie metabolismus MeSH
- vaskulární endoteliální růstové faktory MeSH
- Check Tag
- lidé MeSH
The aim of the present research was to study the uptake of DHEAS, and to establish the intracrine capacity of human platelets to produce sex steroid hormones. The DHEAS transport was evaluated through the uptake of [(3)H]-DHEAS in the presence or absence of different substrates through the organic anion transporting polypeptide (OATP) family. The activity of sulfatase enzyme was evaluated, and the metabolism of DHEAS was measured by the conversion of [(3)H]-DHEAS to [(3)H]-androstenedione, [(3)H]-testosterone, [(3)H]-estrone and [(3)H]-17beta-estradiol. Results indicated the existence in the plasma membrane of an OATP with high affinity for DHEAS and estrone sulphate (E(1)S). The platelets showed the capacity to convert DHEAS to active DHEA by the steroid-sulfatase activity. The cells resulted to be a potential site for androgens production, since they have the capacity to produce androstenedione and testosterone; in addition, they reduced [(3)H]-estrone to [(3)H]-17beta-estradiol. This is the first demonstration that human platelets are able to import DHEAS and E(1)S using the OATP family and to convert DHEAS to active DHEA, and to transform E(1)S to 17beta-estradiol.
- MeSH
- androgeny metabolismus MeSH
- androstendion metabolismus MeSH
- dehydroepiandrosteron metabolismus MeSH
- dehydroepiandrosteronsulfát metabolismus MeSH
- estradiol metabolismus MeSH
- estron analogy a deriváty metabolismus MeSH
- lidé MeSH
- přenašeče organických aniontů metabolismus MeSH
- testosteron metabolismus MeSH
- trombocyty chemie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The recently shown transmissibility of variant Creutzfeldt-Jakob disease (vCJD) by blood transfusion emphasises the need for better understanding of the cellular prion protein (PrPc) in blood. A substantial amount of cell-associated PrPc in blood resides in platelets. Platelet activation leads to up-regulation of PrPc on the platelet surface and its release on exosomes and microparticles. The sub-cellular localisation and function of platelet PrPc, however, is poorly understood. In the present study, we investigated the association of PrPc with platelet lipid rafts and the platelet cytoskeleton. Immuno-fluorescence microscopy showed that the signals of PrPc and P-selectin, both of which occupy intracellular alpha granules, were separated on the membrane, suggesting organisation in different membrane domains. A flotation assay of platelet lysates demonstrated that a relatively small portion of platelet PrPc floats with lipid rafts, regardless of platelet activation status. This was reversed by depolymerisation of the platelet cytoskeleton, which led to flotation of most platelet PrPc, suggesting that interactions with the cytoskeleton prevent flotation of PrPc rafts. This association of PrPc with the platelet cytoskeleton was confirmed by its presence in both the isolated membrane skeleton and actin cytoskeleton. Platelet activation significantly increased the amount of PrPc associated with the cytoskeleton. Our results indicate that the localisation of PrPc in platelets is complex, with the majority of PrPc present within platelet lipid rafts linked to the platelet cytoskeleton. This localisation places PrPc in a position where it can interact with proteins involved in platelet signalling and eventually with vCJD prions.
- MeSH
- aktivace trombocytů MeSH
- biopolymery MeSH
- buněčná membrána chemie MeSH
- cytoplazmatická granula chemie MeSH
- cytoskelet chemie MeSH
- fluorescenční mikroskopie MeSH
- lidé MeSH
- membránové mikrodomény chemie MeSH
- P-selektin MeSH
- PrPC proteiny krev MeSH
- trombocyty chemie ultrastruktura MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
The proteome is the pool of proteins expressed at a given time and circumstance. The word 'proteomics' summarizes several technologies for visualization, quantitation and identification of these proteins. Recent advances in these techniques are helping to elucidate platelet processes which are relevant to bleeding and clotting disorders, transfusion medicine and regulation of angiogenesis. RECENT FINDINGS: Over 1100 platelet proteins have been identified using proteomic techniques. Various subproteomes have been characterized, including platelet releasates (the 'secretome'), alpha and dense granules, membrane and cytoskeletal proteins, platelet-derived microparticles, and the platelet 'phosphoproteome'. Proteomic data about platelets have become increasingly available in integrated databases. SUMMARY: Proteomic experiments in resting and activated platelets have identified novel signaling pathways and secreted proteins which may represent therapeutic targets, as well as potential cancer biomarkers.
- MeSH
- aktivace trombocytů MeSH
- lidé MeSH
- mikropartikule MeSH
- proteom analýza MeSH
- proteomika metody MeSH
- signální transdukce MeSH
- trombocyty chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
5-Hydroxytryptamine (5-HT) can be released from mast cells and platelets through an IgE-dependent mechanism and may play a role in the pathogenesis of allergic bronchoconstriction. However, the effect of 5-HT on ion transport by the airway epithelium is still controversial. The objective of this study was to determine whether 5-hydroxytryptamine (5-HT) regulates NaCl transport by different mechanisms in the apical and basolateral membrane of tracheal epithelia. We studied the rat tracheal epithelium under short-circuit conditions in vitro. Short-circuit current (Isc) was measured in rat tracheal epithelial monolayers cultured on porous filters. 5-HT inhibited Na+ absorption [measured via Na+ short-circuit current (INasc)] in the apical membrane and stimulated Cl- secretion [measured via Cl- short-circuit current (IClsc)] in the basolateral membrane. Functional localization using selective 5-HT agonists and antagonists suggest that IClsc is stimulated by the basolateral membrane-resident 5-HT receptors, whereas INasc is inhibited by the apical membrane-resident 5-HT2 receptors. The basolateral addition of 5-HT increases intracellular cAMP content, but its apical addition does not. The addition of BAPTA/AM blocked the decrease of INasc which was induced by the apical addition of 5-HT, and 5-HT increased intracellular Ca concentrations. These results indicate that 5-HT differentially affects INasc and IClsc across rat tracheal monolayers through interactions with distinct receptors in the apical and the basolateral membrane. These effects may result in an increase of water movement towards the airway lumen.
- MeSH
- bronchospasmus etiologie MeSH
- chlorid sodný chemie metabolismus MeSH
- epitelové buňky cytologie enzymologie chemie MeSH
- imunoglobulin E fyziologie chemie imunologie MeSH
- iontové kanály chemie metabolismus MeSH
- mastocyty cytologie enzymologie chemie MeSH
- potkani Sprague-Dawley MeSH
- serotonin fyziologie chemie MeSH
- trachea cytologie enzymologie chemie MeSH
- trombocyty cytologie enzymologie chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
