Eryptosis is a regulated cell death (RCD) of mature erythrocytes initially described as a counterpart of apoptosis for enucleated cells. However, over the recent years, a growing number of studies have emphasized certain differences between both cell death modalities. In this review paper, we underline the hallmarks of eryptosis and apoptosis and highlight resemblances and dissimilarities between both RCDs. We summarize and critically discuss differences in the impact of caspase-3, Ca2+ signaling, ROS signaling pathways, opposing roles of casein kinase 1α, protein kinase C, Janus kinase 3, cyclin-dependent kinase 4, and AMP-activated protein kinase to highlight a certain degree of divergence between apoptosis and eryptosis. This review emphasizes the crucial importance of further studies that focus on deepening our knowledge of cell death machinery and identifying novel differences between cell death of nucleated and enucleated cells. This might provide evidence that erythrocytes can be defined as viable entities capable of programmed cell destruction. Additionally, the revealed cell type-specific patterns in cell death can facilitate the development of cell death-modulating therapeutic agents.
Membrane surface charge is critical for the transient, yet specific recruitment of proteins with polybasic regions to certain organelles. In eukaryotes, the plasma membrane (PM) is the most electronegative compartment of the cell, which specifies its identity. As such, membrane electrostatics is a central parameter in signaling, intracellular trafficking, and polarity. Here, we explore which are the lipids that control membrane electrostatics using plants as a model. We show that phosphatidylinositol-4-phosphate (PI4P), phosphatidic acidic (PA), and phosphatidylserine (PS) are separately required to generate the electrostatic signature of the plant PM. In addition, we reveal the existence of an electrostatic territory that is organized as a gradient along the endocytic pathway and is controlled by PS/PI4P combination. Altogether, we propose that combinatorial lipid composition of the cytosolic leaflet of organelles not only defines the electrostatic territory but also distinguishes different functional compartments within this territory by specifying their varying surface charges.
- MeSH
- Arabidopsis růst a vývoj metabolismus MeSH
- buněčná membrána metabolismus MeSH
- fosfatidylinositolfosfáty metabolismus MeSH
- fosfatidylseriny metabolismus MeSH
- kořeny rostlin růst a vývoj metabolismus MeSH
- kyseliny fosfatidové metabolismus MeSH
- organely MeSH
- proteiny huseníčku metabolismus MeSH
- signální transdukce MeSH
- statická elektřina * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is a critically important regulatory lipid of the plasma membrane (PM); however, little is known about how cells regulate PM PI(4,5)P2 levels. Here, we show that the phosphatidylinositol 4-phosphate (PI4P)/phosphatidylserine (PS) transfer activity of the endoplasmic reticulum (ER)-resident ORP5 and ORP8 is regulated by both PM PI4P and PI(4,5)P2 Dynamic control of ORP5/8 recruitment to the PM occurs through interactions with the N-terminal Pleckstrin homology domains and adjacent basic residues of ORP5/8 with both PI4P and PI(4,5)P2 Although ORP5 activity requires normal levels of these inositides, ORP8 is called on only when PI(4,5)P2 levels are increased. Regulation of the ORP5/8 attachment to the PM by both phosphoinositides provides a powerful means to determine the relative flux of PI4P toward the ER for PS transport and Sac1-mediated dephosphorylation and PIP 5-kinase-mediated conversion to PI(4,5)P2 Using this rheostat, cells can maintain PI(4,5)P2 levels by adjusting the availability of PI4P in the PM.
- MeSH
- biologický transport MeSH
- buněčná membrána metabolismus MeSH
- endoplazmatické retikulum metabolismus MeSH
- fosfatidylinositol-4,5-difosfát metabolismus MeSH
- fosfatidylinositolfosfáty metabolismus MeSH
- fosfatidylseriny metabolismus MeSH
- fosfotransferasy s alkoholovou skupinou jako akceptorem metabolismus MeSH
- HEK293 buňky MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- proteinové domény MeSH
- steroidní receptory chemie metabolismus MeSH
- substrátová specifita MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, N.I.H., Intramural MeSH
SCOPE: The aim of this work is to identify which proapoptotic pathway is induced in human colon cancer cell lines, in contact with proanthocyanidins extracted from various berries. METHODS AND RESULTS: Proanthocyanidins (Pcys) extracted from 11 berry species are monitored for proapoptotic activities on two related human colon cancer cell lines: SW480-TRAIL-sensitive and SW620-TRAIL-resistant. Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection. Lowbush blueberry extract triggers the strongest activity. When tested on the human monocytic cell line THP-1, blueberry Pcys are less effective for PS externalisation and DNA fragmentation is absent, highlighting a specificity of apoptosis induction in gut cells. In Pcys-treated gut cell lines, caspase 8 (apoptosis extrinsic pathway) but not caspase 9 (apoptosis intrinsic pathway) is activated after 3 hours through P38 phosphorylation (90 min), emphasizing the potency of lowbush blueberry Pcys to eradicate gut TRAIL-resistant cancer cells. CONCLUSION: We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.
- MeSH
- antigeny CD95 metabolismus MeSH
- apoptóza účinky léků MeSH
- brusnice s jedlými plody chemie metabolismus MeSH
- buněčné linie MeSH
- chemorezistence MeSH
- DNA metabolismus MeSH
- fosfatidylseriny metabolismus MeSH
- fosforylace účinky léků MeSH
- fragmentace DNA účinky léků MeSH
- kaspasa 3 metabolismus MeSH
- kaspasa 8 metabolismus MeSH
- kaspasa 9 metabolismus MeSH
- lidé MeSH
- mitogenem aktivované proteinkinasy p38 metabolismus MeSH
- nádory tračníku metabolismus patologie MeSH
- ovoce chemie metabolismus MeSH
- proantokyanidiny chemie izolace a purifikace toxicita MeSH
- protein TRAIL toxicita MeSH
- rostlinné extrakty chemie MeSH
- TRAIL receptory metabolismus MeSH
- Vaccinium vitis-idaea chemie metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Krevní buňky podléhají během odběru, zpracování a skladování významným morfologickým a biochemickým změnám. Struktura a funkce krevních buněk jsou ovlivněny řadou faktorů, počínaje technikou odběru, složením antikoagulačního roztoku, kontaktem s povrchem odběrového vaku či setu, metodou zpracování, obsahem leukocytů v produktu či způsobem deleukotizace. Metabolické a morfologické alterace erytrocytů i trombocytů v koncentrátech limitují jejich skladovatelnost a pravděpodobně mohou ovlivnit terapeutický efekt transfuze. Sdělení předkládá stručný přehled změn, které v krevních buňkách v průběhu skladování probíhají, možnosti monitorování buněčného poškození a shrnuje význam stanovení markerů buněčného poškození pro výzkum, validaci nových postupů a praxi.
Blood cells are subjected to important morphologic and biochemical changes during collection, processing and storage. Their structure and function are influenced by a number of factors, such as collection technique, composition of coagulation solution, contact with the surface of the collection bag or set, method of processing and content of leukocytes in the product or leukodepletion method. Metabolic and morphologic alterations in red blood cells and platelets concentrates limit their storage and may be associated with delayed posttransfusion recovery in vivo. The communication provides a brief summary of changes in blood cells during storage, the possibitity to monitor cell injury and summarizes the significance of markers of cell injury research, validation of new procedures and practice.
Several studies have documented that formation of oxidant mediators may induce apoptosis in nucleated and anucleated cells by modulating intracellular signalling pathways. Reactive oxygen species (ROS) play a very important role in the platelet function. γ-Glutamyltransferase (GGT), a novel source of cellular production of oxidants in the presence of iron and reduced glutathione (GSH), is also found on platelets. The role of platelet-bound GGT in platelet apoptosis and oxidative stress is unknown. The aim of our study was to determine the effects of platelet GGT activity on oxidative stress and apoptotic events in vitro via determination of lipid peroxidation (LPO), protein oxidation, GSH, catalase, caspase-3 activation and phosphatidylserine (PS) exposure in the presence of holo-transferrin (Tf). Stimulation of platelet GGT activity with GSH and glycylglycine (GlyGly) increased caspase-3 activation and PS exposure. A significant increase in lipid and protein oxidation and decrease in GSH and catalase levels was also observed in platelets with stimulation of GGT activity in the presence of Tf. Inhibition of GGT activity effectively reduced all the markers. These results suggest that generation of ROS by the GGT/GSH/Tf system can modify the platelets' redox environment and induce apoptosis in in vitro conditions.
- MeSH
- apoptóza účinky léků MeSH
- dipeptidy metabolismus MeSH
- extracelulární prostor účinky léků metabolismus MeSH
- fosfatidylseriny metabolismus MeSH
- gama-glutamyltransferasa antagonisté a inhibitory metabolismus MeSH
- glutathion metabolismus MeSH
- inhibitory enzymů farmakologie MeSH
- karbonylace proteinů účinky léků MeSH
- kaspasa 3 metabolismus MeSH
- katalasa metabolismus MeSH
- lidé MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- reaktivní formy kyslíku metabolismus MeSH
- sulfhydrylové sloučeniny metabolismus MeSH
- transferin farmakologie MeSH
- trombocyty účinky léků enzymologie patologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Vzhledem k prodlužujícímu se průměrnému věku života je významné předcházet narušení kognitivních funkcí. Jednou z možností jsou správné dietní návyky. Běžná strava nedodává organismu všechny nutné živiny v dostatečném množství. Přitom o takových látkách, jako jsou PUFA, vitaminy B9 a B12 či ginkgo biloba, se předpokládá, že mohou přispět k optimálnímu průběhu stárnutí. Lidé s duševní poruchou konzumují čím dále tím více potravinové doplňky, které mohou mít příznivé i nepříznivé účinky. Je třeba být připraven ke konzultacím na toto téma v běžné klinické praxi.
Owing to increasing average age it is important to prevent impairment of cognitive functions. One of the possibilities is optimal composition of our diet. Our "normal" food is not supplying our organism with essential nutrients in adequate quantities. It is supposed that PUFA, vitamins B9 and B12 or ginkgo biloba are substances which could contribute to optimal course of aging. Individuals with mental disorders are consuming more and more various dietary supplements with possible positive but also negative effects. It is necessary to be prepared for consultations on this topic with patients in our clinical practice.
- MeSH
- dietní záliby psychologie škodlivé účinky MeSH
- fosfatidylseriny metabolismus MeSH
- Ginkgo biloba metabolismus MeSH
- kognitivní poruchy dietoterapie metabolismus prevence a kontrola MeSH
- kyselina listová metabolismus MeSH
- kyseliny mastné omega-3 metabolismus MeSH
- kyseliny mastné omega-6 metabolismus MeSH
- lidé MeSH
- medicína založená na důkazech MeSH
- metaanalýza jako téma MeSH
- potravní doplňky využití MeSH
- rizikové faktory MeSH
- senioři MeSH
- stárnutí fyziologie metabolismus účinky léků MeSH
- stravovací zvyklosti psychologie MeSH
- věkové faktory MeSH
- vitamin B 12 metabolismus MeSH
- vitamin E metabolismus MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
Low-density lipoprotein (LDL) modifications and platelet activation are major risk factors for cardiovascular diseases. When platelets are exposed to oxidative stress, they become activated. Oxidized LDL (ox-LDL) and metal-catalysed oxidation systems such as Fe3+/ascorbic acid increase free radical production. We wanted to verify whether melatonin has a protective effect against oxidative modifications and phosphatidylserine externalization in platelets induced by ox-LDL and Fe3+/ascorbic acid. For in vitro effects of melatonin on platelets, ADPactivated platelets were incubated with ox-LDL or Fe3+/ascorbic acid for 1 h at 37 oC with or without melatonin. Then platelet malondialdehyde, protein carbonyl and glutathione levels were measured. Platelet phosphatidylserine exposure was measured with annexin-V using flow cytometry. Malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with Fe3+/ascorbic acid significantly increased compared to the control group. Glutathione contents of Fe3+/ascorbic acid-treated platelets significantly decreased. Melatonin pre-treatment of Fe3+/ascorbic acid-treated platelets caused a mar ked reduction in malondialdehyde and phosphatidylserine levels and a marked increase in glutathione levels. Melatonin also caused non-significant reduction in protein carbonyl contents of Fe3+/ascorbic acid-treated platelets. Malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with ox-LDL also significantly increased compared to the control group. Platelet glutathione levels non-significantly decreased with ox-LDL. With addition of melatonin, malondialdehyde, protein carbonyl and phosphatidylserine levels of platelets treated with ox-LDL significantly decreased. These data suggest that melatonin may protect platelets from iron overload-induced and ox-LDL-induced oxidative modifications and also from the triggering signals of apoptosis activation, possibly due to its scavenger effect on toxic free radicals.
- MeSH
- antioxidancia farmakologie metabolismus MeSH
- dospělí MeSH
- fosfatidylseriny metabolismus MeSH
- glutathion metabolismus MeSH
- kyselina askorbová farmakologie MeSH
- lidé MeSH
- lipoproteiny LDL farmakologie MeSH
- malondialdehyd metabolismus MeSH
- melatonin farmakologie MeSH
- oxidace-redukce účinky léků MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- trombocyty metabolismus účinky léků MeSH
- železité sloučeniny farmakologie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
We have examined the effect of suberoylanilide hydroxamic acid (SAHA, Vorinostat, Zolinza) on the viability of normal peripheral blood lymphocytes (PBLs) in vitro and on the expression of 20 apoptosis-related genes. RT-PCR, western blots and flow cytometry were performed to reveal the proteins of apoptosis machinery that were affected to cause cell death. Our data suggest that PBL markedly resisted for approximately 24 h the destructive activity of the agent, but eventually 60% of cells treated with 4 micromol/L SAHA died within 72 h through mitochondrial way of apoptosis. While the expression of the majority of genes remained indifferent against 4 micromol/L SAHA, the cellular levels of BimEL, Bmf-2, Bcl-w and survivin mRNA varied, confirming the pro-apoptotic response of SAHA treated PBL. In addition, the expression of multifunctional proteins c-Myc and p21(WAF1) changed profoundly with the time of SAHA treatment. The Bax activator BimEL increased rapidly, driving cells towards apoptosis likely controlled by c-Myc and p21(WAF1) activities. We suggest that variations in c-Myc and p21(WAF1) expression decelerate the apoptosis in the early period and increase the resistance of resting PBL against SAHA. 2009 John Wiley & Sons, Ltd.
- MeSH
- aktivace enzymů účinky léků MeSH
- buněčná membrána metabolismus účinky léků MeSH
- buněčná smrt účinky léků MeSH
- cytochromy c metabolismus MeSH
- cytoprotekce účinky léků MeSH
- financování organizované MeSH
- fosfatidylseriny metabolismus MeSH
- inhibitor p21 cyklin-dependentní kinasy metabolismus MeSH
- kaspasy metabolismus MeSH
- kyseliny hydroxamové farmakologie MeSH
- lidé MeSH
- lymfocyty cytologie enzymologie metabolismus účinky léků MeSH
- membránové proteiny metabolismus MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- mitochondrie metabolismus účinky léků MeSH
- poly(ADP-ribosa)-polymerasy metabolismus MeSH
- proteiny regulující apoptózu metabolismus MeSH
- protoonkogenní proteiny c-myc metabolismus MeSH
- protoonkogenní proteiny metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- regulace genové exprese účinky léků MeSH
- transport proteinů účinky léků MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
The CD8+ natural killer (NK) subpopulation has recently been identified as a fast and reliable biodosimetric indicator within human peripheral blood mononuclear cells (PBMC) in vitro. In irradiated and subsequently cultivated PBMC, a decrease of the relative number of intact CD3-CD8+ lymphocytes 16 and 48 h after treatment has allowed for estimating the received dose in the range of 0 - 10 Gy and lethal/sublethal dose discrimination, respectively. Here we show that suitable biodosimeters can also be found in the peripheral blood B-cell compartment. Multiparameter flow cytometric analysis of irradiated and subsequently cultivated human PBMC revealed that both the CD27+ and CD21- B-cell subpopulations can be used as biodosimeters and the CD19+CD27+ lymphocytes have proved useful for retrospective determination of the received dose in the range of 0 - 6 Gy. In addition, several CD19+ lymphocyte subsets characterized by co-expression of CD21, CD27 and CD38 have been shown to bear biodosimetric potential, too. However, when important parameters like the original size within the CD19+ compartment, its radiation-induced changes and data variation had been taken into account, the CD27+ subpopulation proved superior to the other B-cell subpopulations and subsets. It appears that, in the dose range of 0 - 6 Gy, the relative decrease of CD27+ B lymphocytes provides more sensitive and reliable data than that of CD8+ NK-cells due mainly to lower data variation. In contrast to CD27+ B-cells, the proportions of CD27+ subpopulations of T-cells were not affected by irradiation. We have also proposed a simple experimental protocol based on full blood cultivation and three-color CD27/CD3/CD19 immunophenotyping as a time-saving and inexpensive approach for practical biodosimetric evaluations on simple, three-to-four color flow cytometers.
- Klíčová slova
- B-cells, CD27, Biodosimetric marker, ?-irradiation,
- MeSH
- annexin A5 metabolismus MeSH
- antigeny CD27 fyziologie MeSH
- antigeny CD38 fyziologie MeSH
- B-lymfocyty fyziologie účinky záření MeSH
- biologické markery MeSH
- fenotyp MeSH
- financování organizované MeSH
- fosfatidylseriny metabolismus MeSH
- lidé MeSH
- monocyty fyziologie MeSH
- podskupiny lymfocytů fyziologie MeSH
- průtoková cytometrie MeSH
- receptory buněčného povrchu metabolismus MeSH
- receptory komplementu 3d metabolismus MeSH
- separace buněk MeSH
- vztah dávky záření a odpovědi MeSH
- záření gama MeSH
- Check Tag
- lidé MeSH
