Developing bioinspired materials to convert sunlight into electricity efficiently is paramount for sustainable energy production. Fluorescent proteins are promising candidates as photoactive materials due to their high fluorescence quantum yield and absorption extinction coefficients in aqueous media. However, developing artificial bioinspired photosynthetic systems requires a detailed understanding of molecular interactions and energy transfer mechanisms in the required operating conditions. Here, the supramolecular self-assembly and photophysical properties of fluorescent proteins complexed with organic dyes are investigated in aqueous media. Supercharged mGreenLantern protein, mutated to have a charge of +22, is complexed together with anionic zinc phthalocyanines having 4 or 16 carboxylate groups. The structural characterization reveals a strong electrostatic interaction between the moieties, accompanied by partial conformational distortion of the protein structure, yet without compromising the mGreenLantern chromophore integrity as suggested by the lack of emission features related to the neutral form of the chromophore. The self-assembled biohybrid shows a total quenching of protein fluorescence, in favor of an energy transfer process from the protein to the phthalocyanine, as demonstrated by fluorescence lifetime and ultrafast transient absorption measurements. These results provide insight into the rich photophysics of fluorescent protein-dye complexes, anticipating their applicability as water-based photoactive materials.

• MeSH
• anionty chemie   MeSH
• fluorescenční barviva chemie   MeSH
• fluorescenční spektrometrie MeSH
• indoly * chemie   metabolismus   MeSH
• isoindoly MeSH
• luminescentní proteiny * chemie   metabolismus   MeSH
• organokovové sloučeniny * chemie   metabolismus   MeSH
• přenos energie MeSH
• sloučeniny zinku MeSH
• Publikační typ
• časopisecké články MeSH

UNLABELLED: Methotrexate is used to manage moderate to severe psoriasis and psoriatic arthritis. Methotrexate acts by inhibiting the enzymes involved in nucleotide synthesis. Methotrexate polyglutamates (MTXPGs) have a higher potency to inhibit Dihydrofolate reductase (DHFR), 5-aminoimidazole-4-carboxamide ribonucleotide transformylase (ATIC), and thymidylate synthase (TS), compared to naïve methotrexate. Among all the MTXPGs, methotrexate polyglutamate three (MTXPG-3) is a more potent inhibitor of DHFR, ATIC, and TS enzymes. MTXPG-3 is anticipated to allow therapeutic drug monitoring in immune-mediated inflammatory diseases. We aim to study MTXPG-3 levels as a biomarker for both efficacy and adverse events among psoriatic patients treated with methotrexate monotherapy. We used the LC-MS/MS (Liquid Chromatography Mass Spectrophotometry) system for measuring erythrocyte MTXPG-3. We recruited 106 patients with psoriasis who were treated with methotrexate. Sixty-one of them had psoriatic arthritis (concomitant or in the past). The mean age was 45.08 ± 13.04 years. After twenty-four weeks of methotrexate treatment, 73(69%) were responders, and 33(31%) were non-responders. Thirty-nine (36%) experienced adverse effects, and 67(64%) did not experience any adverse effects. We observed a significant positive correlation between erythrocyte MTXPG-3 and methotrexate dose per week at weeks 12 and 16 but not at week 24. Erythrocyte MTXPG-3 did not correlate with response or adverse effects. It can be used as a marker of compliance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12291-024-01269-x.

• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Depression therapy has been linked to negative effects on energy metabolism, which can be attributed to various factors, including an ongoing inflammatory process commonly seen in metabolic disorders. Unhealthy lifestyle choices of patients and the impact of antidepressants on body weight and lipid and glucose metabolism also contribute to these metabolic side effects. Although not as pronounced as other psychopharmaceuticals, the increasing use of antidepressants raises concerns about their potential impact on public health. The study aimed to evaluate the short- and long-term effects of the antidepressant citalopram and its long-term combination with a special diet on metabolic parameters in mice. METHODS: Animals were randomly divided into 5 groups - control, control + special diet, citalopram (10 mg/kg for 35 days), citalopram + special diet (10 mg/kg for 35 days), and citalopram (10 mg/kg for 7 days). After a described time of administration, animals were anesthetized, blood and fat and liver tissues were collected. Biochemical parameters of lipid metabolism (total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides) and glucose were analyzed using spectrophotometry and relevant adipokines and cytokines were evaluated by ELISA. RESULTS: After a week of application of citalopram, we observed dyslipidemia that persisted even at the end of the 5-week experiment. Furthermore, after 5 weeks of citalopram administration, we observed a significant decrease in body weight gain and decreased leptin levels. Changes in lipid metabolism, higher levels of adipokines leptin and PAI-1 were observed due to the special diet after 5 weeks. CONCLUSIONS: Our research suggests that the effects of citalopram and a diet on the metabolism of mice can be significant, both in the short term (1 week) and in the long term (5 weeks).

• MeSH
• citalopram * škodlivé účinky   aplikace a dávkování   farmakologie   MeSH
• dieta s vysokým obsahem tuků škodlivé účinky   MeSH
• dyslipidemie * chemicky indukované   krev   metabolismus   MeSH
• glukosa * metabolismus   MeSH
• játra metabolismus   účinky léků   MeSH
• krevní glukóza metabolismus   účinky léků   MeSH
• leptin * krev   metabolismus   MeSH
• lipidy * krev   MeSH
• metabolismus lipidů * účinky léků   MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: To evaluate the effect of short-term inhalational exposure to nanoparticles released during dental composite grinding on oxidative stress and antioxidant capacity markers. MATERIALS AND METHODS: Twenty-four healthy volunteers were examined before and after exposure in dental workshop. They spent 76.8 ± 0.7 min in the testing room during grinding of dental nanocomposites. The individual exposure to aerosol particles in each participant ́s breathing zones was monitored using a personal nanoparticle sampler (PENS). Exhaled breath condensate (EBC), blood, and urine samples were collected pre- and post-exposure to measure one oxidative stress marker, i.e., thiobarbituric acid reactive substances (TBARS), and two biomarkers of antioxidant capacity, i.e., ferric-reducing antioxidant power (FRAP) and reduced glutathione (GSH) by spectrophotometry. Spirometry and fractional exhaled nitric oxide (FeNO) were used to evaluate the effect of acute inhalational exposure. RESULTS: Mean mass of dental nanocomposite ground away was 0.88 ± 0.32 g. Average individual doses of respirable particles and nanoparticles measured by PENS were 380 ± 150 and 3.3 ± 1.3 μg, respectively. No significant increase of the post-exposure oxidative stress marker TBARS in EBC and plasma was seen. No decrease in antioxidant capacity biomarkers FRAP and GSH in EBC post-exposure was seen, either. Post-exposure, conjunctival hyperemia was seen in 62.5% volunteers; however, no impairment in spirometry or FeNO results was observed. No correlation of any biomarker measured with individual exposure was found, however, several correlations with interfering factors (age, body mass index, hypertension, dyslipidemia, and environmental pollution parameters) were seen. CONCLUSIONS: This study, using oxidative stress biomarker and antioxidant capacity biomarkers in biological fluids of volunteers during the grinding of dental nanocomposites did not prove a negative effect of this intense short-term exposure. However, further studies are needed to evaluate oxidative stress in long-term exposure of both stomatologists and patients and diverse populations with varying health statuses.

• MeSH
• antioxidancia analýza   MeSH
• biologické markery * analýza   MeSH
• dechové testy MeSH
• dospělí MeSH
• glutathion analýza   MeSH
• inhalační expozice * škodlivé účinky   analýza   MeSH
• látky reagující s kyselinou thiobarbiturovou analýza   MeSH
• lidé MeSH
• nanokompozity * chemie   MeSH
• oxid dusnatý analýza   metabolismus   MeSH
• oxidační stres * MeSH
• pracovní expozice * analýza   škodlivé účinky   MeSH
• zubní lékaři MeSH
• zubní materiály MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: The immunosuppressive roles of galectin-3 (Gal-3) in carcinogenesis make this lectin an attractive target for pharmacological inhibition in immunotherapy. Although current clinical immunotherapies appear promising in the treatment of solid tumors, their efficacy is significantly weakened by the hostile immunosuppressive tumor microenvironment (TME). Gal-3, a prominent TME modulator, efficiently subverts the elimination of cancer, either directly by inducing apoptosis of immune cells or indirectly by binding essential effector molecules, such as interferon-gamma (IFNγ). METHODS: N-(2-Hydroxypropyl)methacrylamide (HPMA)-based glycopolymers bearing poly-N-acetyllactosamine-derived tetrasaccharide ligands of Gal-3 were designed, synthesized, and characterized using high-performance liquid chromatography, dynamic light scattering, UV-Vis spectrophotometry, gel permeation chromatography, nuclear magnetic resonance, high-resolution mass spectrometry and CCK-8 assay for evaluation of glycopolymer non-toxicity. Pro-immunogenic effects of purified glycopolymers were tested by apoptotic assay using flow cytometry, competitive ELISA, and in vitro cell-free INFγ-based assay. RESULTS: All tested glycopolymers completely inhibited Gal-3-induced apoptosis of monocytes/macrophages, of which the M1 subtype is responsible for eliminating cancer cells during immunotherapy. Moreover, the glycopolymers suppressed Gal-3-induced capture of glycosylated IFNγ by competitive inhibition to Gal-3 carbohydrate recognition domain (CRD), which enables further inherent biological activities of this effector, such as differentiation of monocytes into M1 macrophages and repolarization of M2-macrophages to the M1 state. CONCLUSION: The prepared glycopolymers are promising inhibitors of Gal-3 and may serve as important supportive anti-cancer nanosystems enabling the infiltration of proinflammatory macrophages and the reprogramming of unwanted M2 macrophages into the M1 subtype.

• MeSH
• akrylamidy chemie   farmakologie   MeSH
• apoptóza účinky léků   MeSH
• galektin 3 * antagonisté a inhibitory   MeSH
• galektiny MeSH
• interferon gama * metabolismus   MeSH
• krevní proteiny MeSH
• lidé MeSH
• makrofágy účinky léků   MeSH
• monocyty * účinky léků   MeSH
• nádorové mikroprostředí účinky léků   MeSH
• polymery * chemie   farmakologie   MeSH
• protinádorové látky * farmakologie   chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Úvod: Při skladování erytrocytových transfuzních přípravků stoupá hladina volného hemoglobinu, markeru probíhající hemolýzy. Doporučení Rady Evropy (Guide to the preparation, use and quality assurance of blood components, 21st Edition) a česká legislativa (Vyhl. č. 143/2008 Sb.) určují jeho maximální hodnotu jako 0,8 % erytrocytové hmoty na konci doby použitelnosti transfuzního přípravku. Měření % hemolýzy je také doporučeno u erytrocytových transfuzních přípravků, které jsou dále upravovány, např. promytím. Cílem práce bylo porovnat dvě metody měření volného hemoglobinu v supernatantu erytrocytových transfuzních přípravků přímou spektrofotometrií a zjistit vhodnost metody stanovení volného hemoglobinu v transfuzních přípravcích dle Harboea. Soubor a metodika: Do studie bylo zařazeno 20 jednotek erytrocytových transfuzních přípravků. Na konci exspirace byly odebrány vzorky supernatantu a změřena hodnota volného hemoglobinu metodou přímé spektrofotometrie stanovením absorbance vzorku při 540 nm (A) a změřením absorbance při vlnových délkách 415 nm, 450 nm a 380 nm dle Harboea (B). Procento hemolýzy erytrocytární masy pak bylo přepočítáno na objem a hematokrit erytrocytového transfuzního přípravku. Výsledky: Hladina volného hemoglobinu na konci doby skladování erytrocytových transfuzních přípravků nepřesáhla 0,8 % erytrocytové masy u všech jednotek – průměrně 0,31 % (A) a 0,29 % (B). Mezi oběma soubory nebyl zjištěn statisticky ani klinicky významný rozdíl. Závěr: Metoda měření absorbance při vlnových délkách 415 nm, 450 nm a 380 nm dle Harboea odpovídá běžně používané metodě měření při 540 nm a je vhodná k měření obsahu volného hemoglobinu v transfuzních přípravcích.

Introduction: During the storage of erythrocyte transfusion products, the level of free haemoglobin, a marker of ongoing haemolysis, increases. The European Council’s recommendations (Guide to the preparation, use, and quality assurance of blood components, 21st Edition) and Czech legislation (Decree No. 143/2008 Coll.) set the maximum level of free haemoglobin at 0.8% of erythrocyte mass by the end of the transfusion product’s shelf life. Measuring the percentage of haemolysis is also recommended for erythrocyte transfusion products that undergo further processing, such as washing. The aim of this study was to compare two methods for measuring free haemoglobin in the supernatant of erythrocyte transfusion products: direct spectrophotometry and the Harboe method for determining free haemoglobin content in transfusion products. Materials and Methods: The study included 20 units of erythrocyte transfusion products. At the end of the shelf life, supernatant samples were collected, and the free haemoglobin level was measured using direct spectrophotometry by determining the absorbance of the sample at 540 nm (A) and measuring absorbance at wavelengths of 415 nm, 450 nm, and 380 nm according to Harboe (B). The percentage of haemolysis of the erythrocyte mass was then calculated based on the volume and haematocrit of the erythrocyte transfusion product. Results: The level of free haemoglobin at the end of the storage period did not exceed 0.8% of erythrocyte mass for any unit – on average, 0.31% (A) and 0.29% (B). No statistically or clinically significant differences were found between the two methods. Conclusion: The method of measuring absorbance at wavelengths of 415 nm, 450 nm, and 380 nm according to Harboe corresponds to the commonly used method at 540 nm and is suitable for measuring free haemoglobin content in transfusion products.

• MeSH
• erytrocyty * MeSH
• hemolýza * MeSH
• krevní bankovnictví MeSH
• krevní banky * MeSH
• lidé MeSH
• spektrofotometrie metody   MeSH
• Check Tag
• lidé MeSH

Na perfluoralkylové látky (PFAS) je v posledních letech soustředěná pozornost širší veřejnosti, protože to jsou látky znečišťující životní prostředí. Jsou to např. kyselina perfluoroktanová a perfluoroktansulfonová. Tyto látky vznikají průmyslovou aktivitou člověka především při výrobě polymerů nebo nepřilnavých povrchů. Odpadní vodou nebo i jinými cestami se mohou dostat do prostředí, a tak kontaminovat zdroje pitné vody nebo potraviny. Jejich působení na organismy a lidské zdraví je rozsáhle studováno a jejich přítomnosti v organismu je připisován vliv na mnohé zdravotní komplikace, dokonce i některé druhy rakoviny. Z toho důvodu byly stanoveny přípustné limity PFAS v pitné vodě a jejich regulací se zabývají mnohé státní orgány a mezinárodní organizace. Aktuálním standardem v detekci PFAS jsou chromatografické metody. V současnosti jsou zkoumány i nové metody detekce především optickou a elektrochemickou cestou. Jejich příklady jsou v textu detailněji popsány a diskutovány.

Perfluoroalkyl substances (PFAS) have gained wider public attention in recent years as environmental pollutants which include perfluorooctanoic acid and perfluorooctanesulfonic acid. These substances are produced by industry, mainly during the manufacture of polymers or non-stick surfaces. They can enter the environment through waste water or other routes and contaminate drinking water sources or food. Their effects on organisms and human health have been extensively studied and their presence in the body has been attributed to many health complications including cancer. For this reason, limits for PFAS in drinking water have been established and their regulation is being addressed by many governments and international organisations. Chromatographic methods are the current standard for PFAS detection, but new detection methods, mainly optical and electrochemical, are currently being investigated. Examples of these are described in more detail in the text.

• MeSH
• elektrochemické techniky klasifikace   metody   MeSH
• fluorescenční spektrometrie metody   MeSH
• fluorokarbony * analýza   chemie   škodlivé účinky   toxicita   MeSH
• hodnocení vlivů na zdraví metody   MeSH
• impedanční spektroskopie metody   MeSH
• kontaminace potravin MeSH
• látky znečišťující životní prostředí analýza   klasifikace   škodlivé účinky   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Levandule jako rostlina a levandulová silice mají široké spektrum biologických účinků, levandule se v kosmetice a léčivých přípravcích využívá již od dob starého Řecka a Říma. V dnešní době se na trhu vyskytuje nepřeberná škála produktů obsahujících levanduli, ať už jako takovou (levandulový čaj nebo sušenky či jiné pochutiny s květy levandule), případně obsahující levandulovou silici či vodné extrakty levandule (kosmetika, drogerie, potraviny či farmaceutické přípravky). Tato práce se zaměřuje na zjištění obsahu silice, zastoupení těkavých látek a stanovení celkových fenolů a flavonoidů u vzorků levandule lékařské z Levandulového údolí a porovnání výsledků se složením levandulových květů prodávaných jako bylinné čaje v tržní síti České republiky.

Lavender as a herb and the lavender essential oil have a wide range of biological effects. They have been used in cosmetics and medicinal products since the times of ancient Greece and Rome. Nowadays, there is a wide range of products containing lavender on the market, either as such (lavender tea or biscuits or other snacks with lavender flowers) or containing lavender essential oil or aqueous solutions of lavender (cosmetics, drugstore goods, food or pharmaceutical preparations). This work focuses on the determination of the essential oil content, the representation of volatile substances and the determination of total phenols and flavonoids in samples of medicinal lavender from the Levandulové údolí and bought in the Czech shops.

• MeSH
• fenoly analýza   MeSH
• flavonoidy analýza   MeSH
• levandule * chemie   MeSH
• lidé MeSH
• oleje prchavé analýza   MeSH
• spektrofotometrie MeSH
• Check Tag
• lidé MeSH

Changes in the protonation state of lyophilized proteins can impact structural integrity, chemical stability, and propensity to aggregate upon reconstitution. When a buffer is chosen, the freezing/drying process may result in dramatic changes in the protonation state of the protein due to ionization shift of the buffer. In order to determine whether protonation shifts are occurring, ionizable probes can be added to the formulation. Optical probes (dyes) have shown dramatic ionization changes in lyophilized products, but it is unclear whether the pH indicator is uniform throughout the matrix and whether the change in the pH indicator actually mirrors drug ionization changes. In solid-state NMR (SSNMR) spectroscopy, the chemical shift of the carbonyl carbon in carboxylic acids is very sensitive to the ionization state of the acid. Therefore, SSNMR can be used to measure ionization changes in a lyophilized matrix by employing a small quantity of an isotopically-labeled carboxylic acid species in the formulation. This paper compares the apparent pH of six trehalose-containing lyophilized buffer systems using SSNMR and UV-Vis diffuse reflectance spectroscopy (UVDRS). Both SSNMR and UVDRS results using two different ionization probes (butyric acid and bromocresol purple, respectively) showed little change in apparent acidity compared to the pre-lyophilized solution in a sodium citrate buffer, but a greater change was observed in potassium phosphate, sodium phosphate, and histidine buffers. While the trends between the two methods were similar, there were differences in the numerical values of equivalent pH (pHeq) observed between the two methods. The potential causes contributing to the differences are discussed.

• MeSH
• fosfáty * chemie   MeSH
• histidin * chemie   MeSH
• koncentrace vodíkových iontů MeSH
• kyselina citronová chemie   MeSH
• lyofilizace * metody   MeSH
• magnetická rezonanční spektroskopie * metody   MeSH
• pufry MeSH
• spektrofotometrie ultrafialová metody   MeSH
• trehalosa * chemie   MeSH
• Publikační typ
• časopisecké články MeSH

The quantification of cellular metabolic activity via MTT assay has become a widespread practice in eukaryotic cell studies and is progressively extending to bacterial cell investigations. This study pioneers the application of MTT assay to evaluate the metabolic activity of biofilm-forming cells within bacterial biofilms on nanofibrous materials. The biofilm formation of Staphylococcus aureus and Escherichia coli on nanomaterials electrospun from polycaprolactone (PCL), polylactic acid (PLA), and polyamide (PA) was examined. Various parameters of the MTT assay were systematically investigated, including (i) the dissolution time of the formed formazan, (ii) the addition of glucose, and (iii) the optimal wavelength for spectrophotometric determination. Based on interim findings, a refined protocol suitable for application to nanofibrous materials was devised. We recommend 2 h of the dissolution, the application of glucose, and spectrophotometric measurement at 595 nm to obtain reliable data. Comparative analysis with the reference CFU counting protocol revealed similar trends for both tested bacteria and all tested nanomaterials. The proposed MTT protocol emerges as a suitable method for assessing the metabolic activity of bacterial biofilms on PCL, PLA, and PA nanofibrous materials.

• MeSH
• biofilmy * růst a vývoj   MeSH
• Escherichia coli * fyziologie   MeSH
• glukosa metabolismus   MeSH
• nanovlákna * chemie   MeSH
• nylony chemie   MeSH
• polyestery * chemie   MeSH
• spektrofotometrie metody   MeSH
• Staphylococcus aureus * fyziologie   MeSH
• tetrazoliové soli * metabolismus   chemie   MeSH
• thiazoly metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH