To ensure food safety and to prevent unnecessary foodborne complications this study reports fast, fully automated process for histamine determination. This method is based on magnetic separation of histamine with magnetic particles and quantification by the fluorescence intensity change of MSA modified CdSe Quantum dots. Formation of Fe2O3 particles was followed by adsorption of TiO2 on their surface. Magnetism of developed probe enabled rapid histamine isolation prior to its fluorescence detection. Quantum dots (QDs) of approx. 3 nm were prepared via facile UV irradiation. The fluorescence intensity of CdSe QDs was enhanced upon mixing with magnetically separated histamine, in concentration-dependent manner, with a detection limit of 1.6 μM. The linear calibration curve ranged between 0.07 and 4.5 mM histamine with a low LOD and LOQ of 1.6 μM and 6 μM. The detection efficiency of the method was confirmed by ion exchange chromatography. Moreover, the specificity of the sensor was evaluated and no cross-reactivity from nontarget analytes was observed. This method was successfully applied for the direct analysis of histamine in white wine providing detection limit much lower than the histamine maximum levels established by EU regulation in food samples. The recovery rate was excellent, ranging from 84 to 100% with an RSD of less than 4.0%. The main advantage of the proposed method is full automation of the analytical procedure that reduces the time and cost of the analysis, solvent consumption and sample manipulation, enabling routine analysis of large numbers of samples for histamine and highly accurate and precise results.

• MeSH
• Fluorescence MeSH
• Fluorescent Dyes chemistry   MeSH
• Spectrometry, Fluorescence methods   MeSH
• Histamine analysis   MeSH
• Food Contamination analysis   MeSH
• Metal Nanoparticles chemistry   MeSH
• Quantum Dots chemistry   MeSH
• Limit of Detection MeSH
• Magnetic Phenomena MeSH
• Silanes chemistry   MeSH
• Cadmium Compounds chemistry   MeSH
• Tellurium chemistry   MeSH
• Titanium chemistry   MeSH
• Wine analysis   MeSH
• Ferric Compounds chemistry   MeSH
• Publication type
• Journal Article MeSH

STIM1 (stromal interaction molecule 1) and Orai proteins are the essential components of Ca(2+) release-activated Ca(2+) (CRAC) channels. We focused on the role of cholesterol in the regulation of STIM1-mediated Orai1 currents. Chemically induced cholesterol depletion enhanced store-operated Ca(2+) entry (SOCE) and Orai1 currents. Furthermore, cholesterol depletion in mucosal-type mast cells augmented endogenous CRAC currents, which were associated with increased degranulation, a process that requires calcium influx. Single point mutations in the Orai1 amino terminus that would be expected to abolish cholesterol binding enhanced SOCE to a similar extent as did cholesterol depletion. The increase in Orai1 activity in cells expressing these cholesterol-binding-deficient mutants occurred without affecting the amount in the plasma membrane or the coupling of STIM1 to Orai1. We detected cholesterol binding to an Orai1 amino-terminal fragment in vitro and to full-length Orai1 in cells. Thus, our data showed that Orai1 senses the amount of cholesterol in the plasma membrane and that the interaction of Orai1 with cholesterol inhibits its activity, thereby limiting SOCE.

• MeSH
• Biotinylation MeSH
• Point Mutation MeSH
• Cell Membrane metabolism   MeSH
• Cell Line MeSH
• Cholesterol Oxidase metabolism   MeSH
• Cholesterol metabolism   MeSH
• Circular Dichroism MeSH
• Electrophysiological Phenomena MeSH
• Spectrometry, Fluorescence MeSH
• HEK293 Cells MeSH
• Histamine metabolism   MeSH
• Humans MeSH
• Mast Cells metabolism   MeSH
• Mutation MeSH
• Peptides metabolism   MeSH
• Fluorescence Resonance Energy Transfer MeSH
• Signal Transduction MeSH
• Protein Structure, Tertiary MeSH
• Calcium metabolism   MeSH
• Calcium Channels metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• CD8-Positive T-Lymphocytes immunology   drug effects   MeSH
• Cimetidine pharmacology   MeSH
• T-Lymphocytes, Cytotoxic immunology   drug effects   MeSH
• Adult MeSH
• Financing, Organized MeSH
• Histamine pharmacology   MeSH
• HIV Infections genetics   blood   MeSH
• Interferon-gamma isolation & purification   drug effects   MeSH
• Interleukin-2 immunology   MeSH
• Humans MeSH
• Blood Specimen Collection methods   utilization   MeSH
• Perforin isolation & purification   drug effects   MeSH
• Pilot Projects MeSH
• Flow Cytometry methods   utilization   MeSH
• Statistics as Topic MeSH
• Check Tag
• Adult MeSH
• Humans MeSH

Na základě výsledků studií u pacientů s hepatitidami C a malignitami chceme prokázat zvýšení počtu, produkce IFN-gamma a Perforinu HIV-specifickými CD8+ T lymfocyty po inkubaci s Histaminem a IL-2 in vitro. Předpokládáme jejich synergický efekt. Histaminaktivuje cytotoxické CD8+ T lymfocyty (CTL) a zlepšuje jejich funkci, IL-2 je nezbytný pro jejich vyzrávání. Funkce CTL vyšetříme s využitím systému ELISPOT (IFN-gamma)a cytometricky (Perforin). Předpokládaný synergický efekt Histaminu a IL-2 na zlepšení cytotoxických funkcí CD8+ HIV-specifických T lymfocytů vytvoří podmínky pro další studium a může zlepšit prognózu HIV+ pacientů. Jedná se o originální, zatím nepublikovaný návrh na zlepšení cytotoxických funkcí HIV-specifických CD8+ T lymfocytů.; Based on the results of the study with patients with hepatitis C and some oncologic diseases we will study in vitro activation of HIV-specific CD8+ T lymphocytes after incubation with the Histamine, IL-2: their count, production of Perforin and IFN-gammaHistamine induces activation of cytotoxic CD8+ T Lymphocytes and improves their functional activation. IL-2 induces maturation of HIV-specific CD8+ T lymphocytes. We will investigate the production of IFN-gamma by ELISPOT and production of Perforine. Weexpect a synergistic effects of Histamine and IL-2. Enhancement of cytotoxic CD8+ HIV-specific functions should improve the conditions for other experiments and for prognosis of HIV+ patients. It is an original, unpublished project.

• MeSH
• Lymphocyte Activation MeSH
• CD8-Positive T-Lymphocytes MeSH
• T-Lymphocytes, Cytotoxic MeSH
• Hepatitis C immunology   therapy   MeSH
• Histamine therapeutic use   MeSH
• HIV Infections immunology   therapy   MeSH
• Interleukin-2 MeSH
• Perforin MeSH
• Flow Cytometry utilization   MeSH
• Antiretroviral Therapy, Highly Active methods   MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• infekční lékařství
• biochemie
• alergologie a imunologie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

• MeSH
• Spectrometry, Fluorescence MeSH
• Histamine chemistry   MeSH
• Skin pathology   MeSH
• Serotonin chemistry   MeSH