Subunit
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- MeSH
- lidé MeSH
- protilátky virové MeSH
- vakcinace metody MeSH
- vakcíny proti chřipce aplikace a dávkování MeSH
- Check Tag
- lidé MeSH
The vanilloid transient receptor potential channel TRPV3 is a putative molecular thermosensor widely considered to be involved in cutaneous sensation, skin homeostasis, nociception, and pruritus. Repeated stimulation of TRPV3 by high temperatures above 50 °C progressively increases its responses and shifts the activation threshold to physiological temperatures. This use-dependence does not occur in the related heat-sensitive TRPV1 channel in which responses decrease, and the activation threshold is retained above 40 °C during activations. By combining structure-based mutagenesis, electrophysiology, and molecular modeling, we showed that chimeric replacement of the residues from the TRPV3 cytoplasmic inter-subunit interface (N251-E257) with the homologous residues of TRPV1 resulted in channels that, similarly to TRPV1, exhibited a lowered thermal threshold, were sensitized, and failed to close completely after intense stimulation. Crosslinking of this interface by the engineered disulfide bridge between substituted cysteines F259C and V385C (or, to a lesser extent, Y382C) locked the channel in an open state. On the other hand, mutation of a single residue within this region (E736) resulted in heat resistant channels. We propose that alterations in the cytoplasmic inter-subunit interface produce shifts in the channel gating equilibrium and that this domain is critical for the use-dependence of the heat sensitivity of TRPV3.
- MeSH
- cytoplazma metabolismus MeSH
- HEK293 buňky MeSH
- kationtové kanály TRPV chemie genetika metabolismus MeSH
- lidé MeSH
- mutace MeSH
- podjednotky proteinů chemie genetika metabolismus MeSH
- proteinové domény MeSH
- simulace molekulární dynamiky MeSH
- vysoká teplota MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
A complex evaluation of agonist bias at G-protein coupled receptors at the level of G-protein classes and isoforms including non-preferential ones is essential for advanced agonist screening and drug development. Molecular crosstalk in downstream signaling and a lack of sufficiently sensitive and selective methods to study direct coupling with G-protein of interest complicates this analysis. We performed binding and functional analysis of 11 structurally different agonists on prepared fusion proteins of individual subtypes of muscarinic receptors and non-canonical promiscuous α-subunit of G16 protein to study agonist bias. We have demonstrated that fusion of muscarinic receptors with Gα16 limits access of other competitive Gα subunits to the receptor, and thus enables us to study activation of Gα16 mediated pathway more specifically. Our data demonstrated agonist-specific activation of G16 pathway among individual subtypes of muscarinic receptors and revealed signaling bias of oxotremorine towards Gα16 pathway at the M2 receptor and at the same time impaired Gα16 signaling of iperoxo at M5 receptors. Our data have shown that fusion proteins of muscarinic receptors with α-subunit of G-proteins can serve as a suitable tool for studying agonist bias, especially at non-preferential pathways.
- MeSH
- AMP cyklický metabolismus MeSH
- CHO buňky MeSH
- Cricetulus MeSH
- inhibiční koncentrace 50 MeSH
- isoxazoly chemie MeSH
- křečci praví MeSH
- kvartérní amoniové sloučeniny chemie MeSH
- lidé MeSH
- molekulární konformace MeSH
- oxotremorin chemie MeSH
- proteiny vázající GTP - alfa-podjednotky Gq-G11 metabolismus MeSH
- receptory muskarinové metabolismus MeSH
- rekombinantní fúzní proteiny chemie MeSH
- signální transdukce * MeSH
- simulace molekulární dynamiky MeSH
- vazba proteinů MeSH
- zvířata MeSH
- Check Tag
- křečci praví MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Background A trial involving adults 50 years of age or older (ZOE-50) showed that the herpes zoster subunit vaccine (HZ/su) containing recombinant varicella-zoster virus glycoprotein E and the AS01B adjuvant system was associated with a risk of herpes zoster that was 97.2% lower than that associated with placebo. A second trial was performed concurrently at the same sites and examined the safety and efficacy of HZ/su in adults 70 years of age or older (ZOE-70). Methods This randomized, placebo-controlled, phase 3 trial was conducted in 18 countries and involved adults 70 years of age or older. Participants received two doses of HZ/su or placebo (assigned in a 1:1 ratio) administered intramuscularly 2 months apart. Vaccine efficacy against herpes zoster and postherpetic neuralgia was assessed in participants from ZOE-70 and in participants pooled from ZOE-70 and ZOE-50. Results In ZOE-70, 13,900 participants who could be evaluated (mean age, 75.6 years) received either HZ/su (6950 participants) or placebo (6950 participants). During a mean follow-up period of 3.7 years, herpes zoster occurred in 23 HZ/su recipients and in 223 placebo recipients (0.9 vs. 9.2 per 1000 person-years). Vaccine efficacy against herpes zoster was 89.8% (95% confidence interval [CI], 84.2 to 93.7; P<0.001) and was similar in participants 70 to 79 years of age (90.0%) and participants 80 years of age or older (89.1%). In pooled analyses of data from participants 70 years of age or older in ZOE-50 and ZOE-70 (16,596 participants), vaccine efficacy against herpes zoster was 91.3% (95% CI, 86.8 to 94.5; P<0.001), and vaccine efficacy against postherpetic neuralgia was 88.8% (95% CI, 68.7 to 97.1; P<0.001). Solicited reports of injection-site and systemic reactions within 7 days after injection were more frequent among HZ/su recipients than among placebo recipients (79.0% vs. 29.5%). Serious adverse events, potential immune-mediated diseases, and deaths occurred with similar frequencies in the two study groups. Conclusions In our trial, HZ/su was found to reduce the risks of herpes zoster and postherpetic neuralgia among adults 70 years of age or older. (Funded by GlaxoSmithKline Biologicals; ZOE-50 and ZOE-70 ClinicalTrials.gov numbers, NCT01165177 and NCT01165229 .).
- MeSH
- dvojitá slepá metoda MeSH
- herpes zoster imunologie prevence a kontrola MeSH
- Kaplanův-Meierův odhad MeSH
- lidé středního věku MeSH
- lidé MeSH
- postherpetická neuralgie epidemiologie prevence a kontrola MeSH
- riziko MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- subjednotkové vakcíny škodlivé účinky imunologie MeSH
- vakcína proti pásovému oparu * škodlivé účinky imunologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky, fáze III MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
Biogenesis of the plant secondary cell wall involves many important aspects, such as phenolic compound deposition and often silica encrustation. Previously, we demonstrated the importance of the exocyst subunit EXO70H4 for biogenesis of the trichome secondary cell wall, namely for deposition of the autofluorescent and callose-rich cell wall layer. Here, we reveal that EXO70H4-driven cell wall biogenesis is constitutively active in the mature trichome, but also can be activated elsewhere upon pathogen attack, giving this study a broader significance with an overlap into phytopathology. To address the specificity of EXO70H4 among the EXO70 family, we complemented the exo70H4-1 mutant by 18 different Arabidopsis (Arabidopsis thaliana) EXO70 paralogs subcloned under the EXO70H4 promoter. Only EXO70H4 had the capacity to rescue the exo70H4-1 trichome phenotype. Callose deposition phenotype of exo70H4-1 mutant is caused by impaired secretion of PMR4, a callose synthase responsible for the synthesis of callose in the trichome. PMR4 colocalizes with EXO70H4 on plasma membrane microdomains that do not develop in the exo70H4-1 mutant. Using energy-dispersive x-ray microanalysis, we show that both EXO70H4- and PMR4-dependent callose deposition in the trichome are essential for cell wall silicification.
- MeSH
- Arabidopsis účinky léků genetika metabolismus MeSH
- buněčná membrána účinky léků metabolismus MeSH
- buněčná stěna účinky léků metabolismus MeSH
- epidermis rostlin cytologie účinky léků metabolismus MeSH
- fenotyp MeSH
- flagelin farmakologie MeSH
- glukany MeSH
- glukosyltransferasy metabolismus MeSH
- mutace genetika MeSH
- oxid křemičitý metabolismus MeSH
- podjednotky proteinů chemie metabolismus MeSH
- proteinové domény MeSH
- proteiny huseníčku chemie metabolismus MeSH
- regulace genové exprese u rostlin účinky léků MeSH
- trichomy metabolismus MeSH
- up regulace účinky léků MeSH
- vezikulární transportní proteiny chemie metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Complex facial defects resulting from trauma, cancer, or congenital genital disorders present significant challenges for reconstructive surgery. Traditional methods, such as local flaps and grafts, often yield suboptimal aesthetic and functional outcomes. Facial vascularized composite allotransplantation (fVCA) has become a valid option for extensive facial defects. The isolated transplantation of facial subunits, however, potentially allowing for a targeted restoration of smaller defects of individual subunits, is currently not implemented in clinical practice. METHODS: This narrative review synthesizes findings from animal models, human cadaver studies, and clinical experiences to assess the feasibility, anatomical requirements, and immunosuppressive demands of facial subunit transplantation. We examined preclinical studies on vascular supply dynamics and rejection in transplanted tissues, particularly in animal models like nonhuman primates and rats, and cadaver studies focusing on vascularization strategies for facial subunits. RESULTS: Our results indicate that subunit transplantation is anatomically feasible, with established pedicle options for specific regions. Immunosuppression protocols similar to full-face transplantation are required, with preclinical models showing a critical need for optimized immunosuppressive management to prolong graft survival. Cadaver studies reveal that adequate vascularization can be achieved in subunits with the facial artery as the main pedicle. CONCLUSIONS: Facial subunit transplantation offers the potential for improved outcomes in selective facial reconstruction, particularly in functional and aesthetic-critical subunits. However, further advancements in immunosuppression and vascular planning are necessary for clinical application. Addressing these challenges could position subunit transplantation as a less invasive alternative for specific patient populations with tailored benefits regarding localized facial defects.
- MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- obličej * chirurgie MeSH
- předpověď MeSH
- přežívání štěpu MeSH
- transplantace obličeje * metody trendy MeSH
- vaskularizovaná kompozitní alotransplantace * metody trendy MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Transcription factors exert their regulatory potential on RNA polymerase II machinery through a multiprotein complex called Mediator complex or Mediator. The Mediator complex integrates regulatory signals from cell regulatory cascades with the regulation by transcription factors. The Mediator complex consists of 25 subunits in Saccharomyces cerevisiae and 30 or more subunits in multicellular eukaryotes. Mediator subunit 28 (MED28), along with MED30, MED23, MED25 and MED26, belong to presumably evolutionarily new subunits that seem to be absent in unicellular eukaryotes and are likely to have evolved together with multicellularity and cell differentiation. Previously, we have shown that an originally uncharacterized predicted gene, F28F8.5, is the true MED28 orthologue in Caenorhabditis elegans (mdt-28) and showed that it is involved in a spectrum of developmental processes. Here, we studied the proteomic interactome of MDT-28 edited as GFP::MDT-28 using Crispr/Cas9 technology or MDT-28::GFP expressed from extrachromosomal arrays in transgenic C. elegans exploiting the GFPTRAP system and mass spectrometry. The results show that MDT-28 associates with the Head module subunits MDT-6, MDT-8, MDT-11, MDT-17, MDT- 20, MDT-22, and MDT-30 and the Middle module subunit MDT-14. The analyses also identified additional proteins as preferential MDT-28 interactants, including chromatin-organizing proteins, structural proteins and enzymes. The results provide evidence for MDT-28 engagement in the Mediator Head module and support the possibility of physical (direct or indirect) interaction of MDT-28 with additional proteins, reflecting the transcription-regulating potential of primarily structural and enzymatic proteins at the level of the Mediator complex.
- MeSH
- alely MeSH
- Caenorhabditis elegans metabolismus MeSH
- jaderné proteiny metabolismus MeSH
- mediátorový komplex metabolismus MeSH
- podjednotky proteinů metabolismus MeSH
- proteiny Caenorhabditis elegans metabolismus MeSH
- proteomika * MeSH
- vazba proteinů MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Východiska: Mnohočetný myelom je nevyléčitelné onemocnění. Jako standardní terapie je využívána vysokodávkovaná chemoterapie s autologní transplantací kmenových buněk či alogenní transplantací. Relaps onemocnění je však neodvratný, a proto jsou rozvíjeny i jiné směry léčby. Jedním z nich je buněčná imunoterapie, která využívá potenciálu cytotoxických T-lymfocytů. Jako nádorový antigen lze využít nádorově specifické proteiny. Jedním z nich je i katalytická podjednotka telomerázy hTERT a od ní odvozený nonapeptid vázající se na HLA-A2 systém molekul. Typ studie a soubor: Ve studii in vitro byla na souboru zdravých HLA-A2 pozitivních dárců testována možnost aktivace a identifikace myelom-specifických T-lymfocytů s využitím hTERT jako nádorového antigenu. Metody a výsledky: Z mononukleárních buněk periferní krve byly kultivovány T-lymfocyty a dendritické buňky. Dendritické buňky byly pulzovány nonapeptidem hTERT. Po opakované stimulaci T-lymfocytů takto pulzovanými dendritickými buňkami došlo k jejich aktivaci charakterizované produkcí interferonu gama. Závěry: Tato práce ukazuje možnost specifické aktivace a identifikace protinádorových T-lymfocytů, které lze využít při léčbě mnohočetného myelomu.
Backgrounds: Multiple myeloma is an incurable hematological disease. High-dose chemotherapy including autologous stem cell transplantation is recently considered a standard therapy for myeloma. Unfortunately, a relapse of the disease is inevitable. Therefore, new approaches such as immunotherapy have been considered recently. A specific activation of cytotoxic T cells can be reached using dendritic cells loaded with tumor-specific antigen. Catalytic subunit of telomerase hTERT and an HLA-A2-specific nonapeptide derived from hTERT can be used. Design and subjects: Activation and identification of myeloma-specific T cells from healthy HLA-A2 blood donors has been tested in an in vitro study using hTERT-derived nonapeptide as a tumor-specific antigen. Methods and results: T cells and dendritic cells were obtained from peripheral blood. T cells were repeatedly stimulated with hTERT nonapeptide- loaded dendritic cells. Activated myeloma-specific T cells produced interferon gamma and were evaluated by flow cytometry. Conclusion: This study demonstrates feasibility of an in vitro identification of tumor-specific T cells that can be used in myeloma therapy.
