Protein cross-linking has assumed an irreplaceable role in structural proteomics. Recently, significant efforts have been made to develop novel mass spectrometry (MS)-cleavable reagents. At present, only water-insoluble MS-cleavable cross-linkers are commercially available. However, to comprehensively analyse the various chemical and structural motifs making up proteins, it is necessary to target different protein sites with varying degrees of hydrophilicity. Here we introduce the new MS-cleavable cross-linker disulfodisuccinimidyl dibutyric urea (DSSBU), which we have developed in-house for this purpose. DSSBU contains an N-hydroxysulfosuccinimide (sulfo-NHS) reactive group, so it can serve as a water-soluble counterpart to the widely used cross-linker disuccinimidyl dibutyric urea (DSBU). To investigate the applicability of DSSBU, we compared the efficacy of four similar cross-linkers: bis[sulfosuccinimidyl] suberate (BS3), disuccinimidyl suberate (DSS), DSBU and DSSBU with bovine serum albumin. In addition, we compared the efficacy of DSBU and DSSBU with human haemoglobin. Our results demonstrate that the sulfo-NHS group ensures the superior water solubility of DSSBU and thus negates the need for organic solvents such as dimethyl sulfoxide while preserving the effectivity of urea-based MS-cleavable crosslinkers such as DSBU. Additionally, it makes it possible to target polar regions in proteins. The data gathered are available via ProteomeXchange under identifier PXD055284. SIGNIFICANCE: We have synthesized the novel protein cross-linker DSSBU, which combines sulfo-NHS ester chemistry with a mass spectrometry-cleavable urea group. This makes DSSBU a water-soluble, MS-cleavable cross-linker that reacts with amino groups. To our knowledge, it is the first cross-linker which combines all three of these characteristics. We have tested the performance of our novel cross-linker on bovine serum albumin, a model widely used by the cross-linking mass spectrometry community, and on human haemoglobin. We have comprehensively assessed the performance of DSSBU and compared its efficacy with that of three other cross-linkers in current use (BS3, DSS and DSBU). We conclude that our novel cross-linker surpasses its MS-non-cleavable analogue BS3 in performance and that its water solubility eliminates the need for organic solvents while its hydrophilicity allows for the targetting of polar regions in proteins. Therefore, it will likely become a significant addition to the portfolio of N-hydroxysuccinimide ester cross-linkers.
- MeSH
- hmotnostní spektrometrie metody MeSH
- lidé MeSH
- močovina chemie MeSH
- proteomika metody MeSH
- reagencia zkříženě vázaná * chemie MeSH
- sérový albumin hovězí chemie MeSH
- skot MeSH
- sukcinimidy * chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Myricetin (MYR) and ampelopsin (AMP, or dihydromyricetin) are flavonoid aglycones found in certain plants and dietary supplements. During the presystemic biotransformation of flavonoids, mainly sulfate and glucuronide derivatives are produced, which are the dominant metabolites in the circulation. In this study, we tested the interactions of MYR, myricetin-3'-O-sulfate (M3'S), AMP, and ampelopsin-4'-O-sulfate (A4'S) with human serum albumin (HSA), cytochrome P450 enzymes (CYPs), and organic anion-transporting polypeptides (OATPs) using in vitro models, including the recently developed method for measuring flavonoid levels in living cells. M3'S and MYR bound to albumin with high affinity, and they showed moderate displacing effects versus the Site I marker warfarin. MYR, M3'S, AMP, and A4'S exerted no or only minor inhibitory effects on CYP2C9, CYP2C19, and CYP3A4 enzymes. M3'S and MYR caused considerable inhibitory actions on OATP1B1 at low micromolar concentrations (IC50 = 1.7 and 6.4 μM, respectively), while even their nanomolar levels resulted in strong inhibitory effects on OATP2B1 (IC50 = 0.3 and 0.4 μM, respectively). In addition, M3'S proved to be a substrate of OATP1B1 and OATP2B1. These results suggest that MYR-containing dietary supplements may affect the OATP-mediated transport of certain drugs, and OATPs are involved in the tissue uptake of M3'S.
- MeSH
- cytochrom P-450 CYP3A metabolismus MeSH
- cytochrom P450 CYP2C9 metabolismus MeSH
- flavonoidy * farmakologie MeSH
- flavonoly farmakologie MeSH
- lidé MeSH
- polypeptid C přenášející organické anionty * metabolismus MeSH
- přenašeče organických aniontů * metabolismus MeSH
- sérový albumin metabolismus MeSH
- sírany metabolismus MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Several novel copper (II) complexes of reduced Schiff bases containing fluoride substituents were prepared and structurally characterized by single-crystal X-ray diffraction. The complexes exhibited diverse structures, with the central atom in distorted tetrahedral geometry. The biological effects of the products were evaluated, specifically their cytotoxicity, antimicrobial, and antiurease activities, as well as affinity for albumin (BSA) and DNA (ct-DNA). The complexes showed marked cytotoxic activities in the HepG2 hepatocellular carcinoma cell line, considerably higher than the standard cisplatin. The cytotoxicity depended significantly on the substitution pattern. The best activity was observed in the complex with a trifluoromethyl group in position 4 of the benzene ring-the dichloro[(±)-trans-N,N'-bis-(4-trifluoromethylbenzyl)-cyclohexane-1,2-diamine]copper (II) complex, whose activity (IC50 28.7 μM) was higher than that of the free ligand and markedly better than the activity of the standard cisplatin (IC50 336.8 μM). The same complex also showed the highest antimicrobial effect in vitro. The affinity of the complexes towards bovine serum albumin (BSA) and calf thymus DNA (ct-DNA) was established as well, indicating only marginal differences between the complexes. In addition, all complexes were shown to be excellent inhibitors of the enzyme urease, with the IC50 values in the lower micromolar region.
- MeSH
- buňky Hep G2 MeSH
- DNA metabolismus chemie MeSH
- fluor chemie MeSH
- komplexní sloučeniny * farmakologie chemie chemická syntéza MeSH
- lidé MeSH
- ligandy MeSH
- měď * chemie MeSH
- protinádorové látky * farmakologie chemie MeSH
- Schiffovy báze * chemie farmakologie MeSH
- sérový albumin hovězí chemie MeSH
- ureasa antagonisté a inhibitory metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Total hip (THA) or knee (TKA) arthroplasty is still a traumatic and challenging operation that induces inflammation, with a particularly high risk of acute-phase reaction. The aim of this study was to predict the likelihood of implant-associated complications during the preoperative and postoperative course. METHODS: The prospective observational, non-interventional study of patients diagnosed with primary knee or hip osteoarthrosis undergoing THA or TKA during the study period was conducted. The inflammatory and malnutrition parameters were collected for each patient one day before surgery, two days after surgery, and in outpatient follow-up. RESULTS: Of 159 patients analysed, 12 developed implant-associated complications. The albumin, prealbumin, Intensive Care Infection Score (ICIS), Nutritional Risk Index, and white blood cell counts were found to be potential predictors. Notably, preoperative albumin levels significantly differed between groups with and without complications (P-value = 0.042). CONCLUSION: Our study definitively shows that WBC, prealbumin, Nutritional Risk Index, ICIS as a novel marker, and significantly albumin, outperform C-reactive protein in predicting implant-associated complications in hip and knee arthroplasty.
- MeSH
- artróza kyčelních kloubů chirurgie MeSH
- biologické markery krev MeSH
- C-reaktivní protein metabolismus analýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- náhrada kyčelního kloubu * škodlivé účinky MeSH
- pooperační komplikace * etiologie krev MeSH
- prealbumin metabolismus analýza MeSH
- prospektivní studie MeSH
- rizikové faktory MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- sérový albumin analýza metabolismus MeSH
- totální endoprotéza kolene * škodlivé účinky 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
- pozorovací studie MeSH
Transthyretinová amyloidóza je multisystémové onemocnění s variabilní manifestací a poměrně nepříznivou prognózou. Novou možností specifické terapie je tafamidis, selektivní stabilizátor transthyretinu. V současné době je tafamidis indikován k léčbě jak wild-type, tak i hereditární formy onemocnění s cílem redukce symptomů, hospitalizací z kardiovaskulárních příčin a mortality u pacientů, kteří jsou ve funkční třídě NYHA I nebo II.
Transthyretin amyloidosis is a multisystem disease with variable manifestations and a relatively unfavorable prognosis. A new option for specific therapy is tafamidis, a selective transthyretin stabilizer. Currently, tafamidis is indicated for the treatment of wild-type and hereditary forms of the disease with the goal of reducing symptoms, cardiovascular hospitalizations and mortality in patients who are in NYHA functional class I or II.
- Klíčová slova
- transthyretinová amyloidóza, tafamidis, studie ATTR-ACT,
- MeSH
- amyloidóza * farmakoterapie genetika mortalita patofyziologie MeSH
- časná diagnóza MeSH
- kardiomyopatie diagnóza etiologie mortalita MeSH
- lidé MeSH
- polyneuropatie diagnóza etiologie MeSH
- prealbumin antagonisté a inhibitory fyziologie MeSH
- randomizované kontrolované studie jako téma MeSH
- věkové faktory MeSH
- Check Tag
- lidé MeSH
- MeSH
- biologické markery * krev MeSH
- dítě MeSH
- kreatinin krev MeSH
- lidé MeSH
- lidský sérový albumin analýza klasifikace MeSH
- podvýživa * diagnóza klasifikace krev MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- přehledy MeSH
Výživa významně ovlivňuje zdravotní stav jedince, protože ovlivňují biochemické a energetické děje v těle. Děti jsou k důsledkům poruch výživy významně náchylnější než dospělí. Zhodnocení nutričního stavu pacienta vychází z anamnestických údajů a výsledků fyzikálních, laboratorních, funkčních a zobrazovacích vyšetření. Hodnocení a interpretace laboratorních nálezů bývají obtížné, ale na druhou stranu jsou pro odhad nutriční bilance stěžejní. Proto se předkládaný článek věnuje interpretaci nejčastěji užívaných nutričních laboratorních parametrů a upozorňuje na některá jejich úskalí a souvislosti z pohledu patofyziologie.
Nutrition significantly affects the health of an individual because it influences the biochemical and energy processes in the body. Children are significantly more susceptible to the effects of nutritional disorders than adults. The assessment of a patient’s nutritional status is based on anamnestic data and the results of physical, laboratory, functional and imaging tests. The evaluation and interpretation of laboratory findings is often difficult, but on the other hand is central to the estimation of nutritional balance. Therefore, the present article focuses on the interpretation of the most commonly used nutritional laboratory parameters and highlights some of their pitfalls and implications in terms of pathophysiology.
- MeSH
- avitaminóza patologie MeSH
- biologické markery * MeSH
- dítě MeSH
- hodnocení stavu výživy * MeSH
- karenční nemoci etiologie patofyziologie patologie MeSH
- lidé MeSH
- podvýživa diagnóza MeSH
- prealbumin analýza MeSH
- sběr dat metody MeSH
- sérový albumin analýza MeSH
- stopové prvky analýza nedostatek MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- přehledy MeSH
Drug binding to plasma proteins influences processes such as liberation, adsorption, disposition, metabolism, and elimination of drugs, which are thus one of the key steps of a new drug development. As a result, the characterization of drug-protein interactions is an essential part of these time- and money-consuming processes. It is important to determine not only the binding strength and the stoichiometry of interaction, but also the binding site of a drug on a protein molecule, because two drugs with the same binding site can mutually affect free drug concentration. Capillary electrophoresis-frontal analysis with mobility shift affinity capillary electrophoresis is one of the most used affinity capillary electrophoresis methods for the characterization of these interactions. In this study, a well-known sensitivity problem of most capillary electrophoresis-frontal analyses using ultraviolet detection is solved by its combination with contactless conductivity detection, which provided sixfold lower limits of quantitation and detection. Binding parameters of the human serum albumin-salicylic acid model affinity pair were evaluated by this newly developed approach and by the classical approach with ultraviolet detection primarily used for their mutual comparison. The results of both approaches agreed well and are also in agreement with literature data obtained using different techniques.
- MeSH
- elektrická vodivost MeSH
- elektroforéza kapilární metody MeSH
- krevní proteiny * MeSH
- lidé MeSH
- lidský sérový albumin * MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Surface and treated wastewater are contaminated with highly complex mixtures of micropollutants, which may cause numerous adverse effects, often mediated by endocrine disruption. However, there is limited knowledge regarding some important modes of action, such as interference with thyroid hormone (TH) regulation, and the compounds driving these effects. This study describes an effective approach for the identification of compounds with the potential to bind to transthyretin (TTR; protein distributing TH to target tissues), based on their specific separation in a pull-down assay followed by non-target analysis (NTA). The method was optimized with known TTR ligands and applied to complex water samples. The specific separation of TTR ligands provided a substantial reduction of chromatographic features from the original samples. The applied NTA workflow resulted in the identification of 34 structures. Twelve compounds with available standards were quantified in the original extracts and their TH-displacement potency was confirmed. Eleven compounds were discovered as TTR binders for the first time and linear alkylbenzene sulfonates (LAS) were highlighted as contaminants of concern. Pull-down assay combined with NTA proved to be a well-functioning approach for the identification of unknown bioactive compounds in complex mixtures with great application potential across various biological targets and environmental compartments.
OBJECTIVES: Acute intestinal ischemia is a severe complication of abdominal aortic surgery that is difficult to diagnose early and therefore to treat adequately and timely. In this study the perioperative kinetics of d-lactate and ischemia-modified albumin (IMA) are described and the predictive value of these markers for the early diagnosis of acute intestinal ischemia is assessed. DESIGN & METHODS: This non-randomised, single-centre cohort study enrolled 50 patients with abdominal aortic aneurysm (AAA) and 30 patients with aortoiliac occlusive disease (AOID). Serum d-lactate and IMA were assessed pre-, intra-, and postoperatively at eight defined time points. RESULTS: The highest serum d-lactate was at 6 h after complete declamping of the vascular graft. The highest predictive power of d-lactate was at 3 h after complete declamping (AUC 0.857). IMA was found to be higher in the AAA group in ischemic patients 10 min after complete declamping than in the AOID group. The highest predictive values of IMA were at 1 h after aortic cross-clamping (AUC 0.758) and 3 and 6 h after complete declamping (0.745 and 0.721, respectively). Moreover, the multivariate model with both markers at 3 h after complete declamping improved the detection of intestinal ischemia (AUC 0.894). CONCLUSIONS: Serum levels of IMA and d-lactate seem to be influential predictive markers for postoperative intestinal ischemia, especially after 3 h from complete declamping of vascular reconstruction.
- MeSH
- aneurysma břišní aorty * chirurgie komplikace MeSH
- biologické markery MeSH
- ischemie diagnóza etiologie MeSH
- kohortové studie MeSH
- kyselina mléčná * MeSH
- lidé MeSH
- sérový albumin MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
