Colorimetry is a widely used technique for optical detection in point-of-care testing and on-site detection. Although some studies employ a multiplex approach to analyse coloured solutions, many still analyse one sample at a time. We have prepared a simple and affordable colorimetric assay based on a TCS34725 colour sensor (ams-OSRAM) integrated into an M5Stack module and an RGB LED module both inserted into a 3D printed frame. We found that the colorimetric assay can be easily transferred to a colour sensing platform, and the signal range obtained using the prepared colorimeter is more than 200 times larger than that obtained using digital image colorimetry (DIC) for the same samples containing cholinesterase or urease as a model enzyme providing a change in pH of the processed solution. The assay appears to be ready for practical use.

• MeSH
• kolorimetrie * přístrojové vybavení   metody   MeSH
• koncentrace vodíkových iontů MeSH
• ureasa chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ureasa 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.

• Klíčová slova
• BSA binding, DNA binding, anticancer activity, antimicrobial activity, copper, cytotoxicity, metal complexes, urease inhibition,
• 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
• Názvy látek
• calf thymus DNA MeSH Prohlížeč
• DNA MeSH
• fluor MeSH
• komplexní sloučeniny * MeSH
• ligandy MeSH
• měď * MeSH
• protinádorové látky * MeSH
• Schiffovy báze * MeSH
• sérový albumin hovězí MeSH
• ureasa MeSH

AIMS: The aim of the present research was to synthesize glycoluril derivative 2,4-Bis(4- cyanobenzyl)glycoluril through a convergent scheme. BACKGROUND: For this purpose, Sandmeyer reaction procedure was employed for the synthesis of said compound. The structure of the pure compound was confirmed by using different spectroscopic techniques, such as 1HNMR, 13C-NMR and (HR-MS) Mass spectrometry. OBJECTIVE: Convergent synthesis of 2,4-BIS (4-CYANOBENZYL)GLYCOLURIL USING SANDMEYER REACTION and urease inhibition study. METHODS: The structure of the pure compound was confirmed by using different spectroscopic techniques such as 1H-NMR, 13C-NMR and (HR-MS) Mass spectrometry. The electronic properties of the newly synthesized compound and thiourea were determined by using density functional theory. RESULTS: Furthermore, the compound was evaluated against urease enzyme and was found to be potent inhibitors with an IC50 value of 11.5 ± 1.50 μM when compared with standard inhibitor thiourea (IC50 = 21.0 ± 1.90 μM). The compound may serve as a lead compound to synthesize new cyano-based bambusuril in the future with enhanced biological properties. CONCLUSION: We have synthesized a new glycoluril derivative 2,4-Bis(4-cyanobenzyl)glycoluril by the sandmeyer reaction. It has been obtained in the form of light yellowish powder in good yield (96%). Glycoluril based macrocycles have been used in various fields; starting from the 2,4-Bis(4-nitrobenzyl)glycoluril (already reported compound), which has undergone reduction (CH3OH,Pt/C) , diazotization (NaNO2/HCl), cyanation (CuCl/KCN), respectively in order to synthesize the desired new glycoluril derivative. The obtained product will be used as a building block for the synthesis of the cyano based bambusuril marcocycle in the future. The yield of the obtained product has been monitored by using different amounts of cyanating reagent, but the best results are shown by the use of 4 mmol of CuCl/KCN. KCN with CuCl assisted the conversion of diazo group into the cyano group with enhanced yield when used in excess amount. It acts as a catalyst. The solubility characteristic of 2,4-Bis(4-cyanobenzyl)glycoluril has also been determined in different organic solvents. 1H NMR technique proved to be very helpful for the structure determination of our desired product. Benzylic protons give signals at 7.5 ppm and 7.8 ppm, respectively. The downfield peaks confirm the presence of CN group near the benzylic protons. Methine protons show a signal at 5.2 ppm, which ensures the basic skeleton of glycoluril. Ureidyl protons also confirm the synthesis of the heterocyclic 2,4-Bis(4-cyanobenzyl)glycoluril compound. The negative and positive electrostatic potential sites, molecular descriptors, and charge density distribution of frontier molecular orbitals are revealing that 4a with promising sites for electrophilic and nucleophilic attacks would result to enhance the urease inhibition, which is in good agreement with the experimental data.

• Klíčová slova
• (HR-MS) mass spectrometry, 1H NMR, Glycoluril derivative, IC50, bambusuril., density functional theory (DFT), sandmeyer reaction, thiourea, urease enzyme,
• MeSH
• heterocyklické sloučeniny bicyklické MeSH
• imidazolidiny MeSH
• imidazoly MeSH
• inhibitory enzymů * farmakologie   MeSH
• simulace molekulového dockingu MeSH
• teorie funkcionálu hustoty MeSH
• ureasa * metabolismus   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glycoluril MeSH Prohlížeč
• heterocyklické sloučeniny bicyklické MeSH
• imidazolidiny MeSH
• imidazoly MeSH
• inhibitory enzymů * MeSH
• ureasa * MeSH

In the current study, pyroglutamic acid (pGlu), a natural amino acid derivative, has efficiently inhibited the catalytic activities of three important enzymes, namely: Human recombinant phosphodiesterase-5A1 (PDE5A1), human angiotensin-converting enzyme (ACE), and urease. These enzymes were reported to be associated with several important clinical conditions in humans. Radioactivity-based assay, spectrophotometric-based assay, and an Electrospray Ionization-Mass Spectrometry-based method were employed to ascertain the inhibitory actions of pGlu against PDE5A1, ACE, and urease, respectively. The results unveiled that pGlu potently suppressed the activity of PDE5A1 (half-maximal inhibitory concentration; IC50 = 5.23 µM) compared with that of standard drug sildenafil citrate (IC50 = 7.14 µM). Moreover, pGlu at a concentration of 20 µg/mL was found to efficiently inhibit human ACE with 98.2% inhibition compared with that of standard captopril (99.6%; 20 µg/mL). The urease-catalyzed reaction was also remarkably inactivated by pGlu and standard acetohydroxamic acid with IC50 values of 1.8 and 3.9 µM, respectively. Remarkably, the outcome of in vitro cytotoxicity assay did not reveal any significant cytotoxic properties of pGlu against human cervical carcinoma cells and normal human fetal lung fibroblast cells. In addition to in vitro assays, molecular docking analyses were performed to corroborate the outcomes of in vitro results with predicted structure-activity relationships. In conclusion, pGlu could be presented as a natural and multifunctional agent with promising applications in the treatment of some ailments connected with the above-mentioned anti-enzymatic properties.

• Klíčová slova
• ESI-mass spectrometry, angiotensin-converting enzyme, anti-enzymatic properties, cytotoxicity, phosphodiesterase 5, pyroglutamic acid, urease,
• MeSH
• angiotensin konvertující enzym chemie   genetika   metabolismus   MeSH
• buněčné linie MeSH
• cyklické nukleotidfosfodiesterasy, typ 5 chemie   genetika   metabolismus   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
• inhibiční koncentrace 50 MeSH
• kaptopril chemie   metabolismus   MeSH
• kyselina pyrrolidonkarboxylová chemie   metabolismus   toxicita   MeSH
• kyseliny hydroxamové antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• rekombinantní proteiny biosyntéza   chemie   izolace a purifikace   MeSH
• sildenafil citrát chemie   metabolismus   MeSH
• simulace molekulového dockingu MeSH
• spektrofotometrie MeSH
• terciární struktura proteinů MeSH
• ureasa antagonisté a inhibitory   metabolismus   MeSH
• vazebná místa MeSH
• viabilita buněk účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ACE protein, human MeSH Prohlížeč
• acetohydroxamic acid MeSH Prohlížeč
• angiotensin konvertující enzym MeSH
• cyklické nukleotidfosfodiesterasy, typ 5 MeSH
• kaptopril MeSH
• kyselina pyrrolidonkarboxylová MeSH
• kyseliny hydroxamové MeSH
• PDE5A protein, human MeSH Prohlížeč
• rekombinantní proteiny MeSH
• sildenafil citrát MeSH
• ureasa MeSH

The increasing resistance of pathogens to common antibiotics, as well as the need to control urease activity to improve the yield of soil nitrogen fertilization in agricultural applications, has stimulated the development of novel classes of molecules that target urease as an enzyme. In this context, the newly developed compounds on the basis of 1-heptanoyl-3-arylthiourea family were evaluated for Jack bean urease enzyme inhibition activity to validate their role as potent inhibitors of this enzyme. 1-Heptanoyl-3-arylthioureas were obtained in excellent yield and characterized through spectral and elemental analysis. All the compounds displayed remarkable potency against urease inhibition as compared to thiourea standard. It was found that novel compounds fulfill the criteria of drug-likeness by obeying Lipinski's rule of five. Particularly compound 4a and 4c can serve as lead molecules in 4D (drug designing discovery and development). Kinetic mechanism and molecular docking studies also carried out to delineate the mode of inhibition and binding affinity of the molecules.

• Klíčová slova
• Acyl thioureas, Antioxidant, Jack bean urease, Kinetic mechanism, Lipinski’s rules, Molecular modeling, Urease,
• MeSH
• Canavalia enzymologie   MeSH
• inhibitory enzymů chemie   farmakologie   MeSH
• kinetika MeSH
• molekulární struktura MeSH
• simulace molekulového dockingu * MeSH
• thiomočovina chemie   farmakologie   MeSH
• ureasa antagonisté a inhibitory   metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• inhibitory enzymů MeSH
• thiomočovina MeSH
• ureasa MeSH

A bistable switch from a low pH (unreacted "off") state to a high pH (reacted "on") state was obtained in enzyme-loaded gel beads in response to supra-threshold substrate concentrations.

• MeSH
• algináty chemie   metabolismus   MeSH
• difuze MeSH
• koncentrace vodíkových iontů MeSH
• kyselina glukuronová chemie   metabolismus   MeSH
• kyseliny hexuronové chemie   metabolismus   MeSH
• ureasa chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• algináty MeSH
• kyselina glukuronová MeSH
• kyseliny hexuronové MeSH
• ureasa MeSH

It was originally shown [10] that urease retains its enzymatic activity when adsorbed at bare mercury and solid amalgam surfaces. However the opinion later prevailed that, when adsorbed at bare metal electrodes, proteins are irreversibly denatured. Here we confirm that urease is enzymatically active at a bare solid amalgam surface as found by Santhanam et al., and we show that this enzyme is equally active at a thiol-modified amalgam surface. We also show that it is the reduced form of urease, which is enzymatically active at Hg surfaces. Oxidation of the protein, resulting in formation of disulfide bonds, strongly decreases the enzyme activity. Using constant current chronopotentiometric stripping (CPS) we show that the exposure of surface-attached urease to negative potentials results in the protein unfolding. The extent of the unfolding depends upon the amount of time for which the protein is exposed to negative potentials, and at very short times this unfolding can be avoided. At thiol-modified Hg surfaces the protein is less vulnerable to the effects of the electric field. We conclude that the loss of enzymatic activity, resulting from a 10 min exposure of the protein to -0.58 V, is not due to reduction of the disulfide bonds as suggested by Santhanam et al. This loss is probably a result of protein reorientation, due to reduction of the Hg-S bonds (formed by accessible cysteines), followed by prolonged electric field effect on the surface-attached protein.

• Klíčová slova
• Constant-current chronopotentiometric stripping, Mercury containing electrodes, Protein denaturation at negatively charged surfaces, Protein structure at surfaces, Thiol-modified electrodes, Urease enzymatic activity,
• MeSH
• adsorpce MeSH
• cystein chemie   MeSH
• denaturace proteinů MeSH
• disulfidy chemie   MeSH
• dithiothreitol chemie   MeSH
• elektrochemické techniky MeSH
• elektrody MeSH
• katalýza MeSH
• oxidace-redukce MeSH
• povrchové vlastnosti MeSH
• rtuť chemie   MeSH
• sbalování proteinů MeSH
• sulfhydrylové sloučeniny chemie   MeSH
• teplota MeSH
• ureasa chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cystein MeSH
• disulfidy MeSH
• dithiothreitol MeSH
• rtuť MeSH
• sulfhydrylové sloučeniny MeSH
• ureasa MeSH

More than 40% of nosocomial infections are those of the urinary tract, most of these occurring in catheterized patients. Bacterial colonization of the urinary tract and catheters results not only in infection, but also various complications, such as blockage of catheters with crystalline deposits of bacterial origin, generation of gravels and pyelonephritis. The diversity of the biofilm microbial community increases with duration of catheter emplacement. One of the most important pathogens in this regard is Proteus mirabilis. The aims of this study were to identify and assess particular virulence factors present in catheter-associated urinary tract infection (CAUTI) isolates, their correlation and linkages: three types of motility (swarming, swimming and twitching), the ability to swarm over urinary catheters, biofilm production in two types of media, urease production and adherence of bacterial cells to various types of urinary tract catheters. We examined 102 CAUTI isolates and 50 isolates taken from stool samples of healthy people. Among the microorganisms isolated from urinary catheters, significant differences were found in biofilm-forming ability and the swarming motility. In comparison with the control group, the microorganisms isolated from urinary catheters showed a wider spectrum of virulence factors. The virulence factors (twitching motility, swimming motility, swarming over various types of catheters and biofilm formation) were also more intensively expressed.

• MeSH
• bakteriální adheze MeSH
• biofilmy růst a vývoj   MeSH
• faktory virulence metabolismus   MeSH
• infekce bakteriemi rodu Proteus mikrobiologie   MeSH
• infekce močového ústrojí mikrobiologie   MeSH
• katétrové infekce mikrobiologie   MeSH
• lidé MeSH
• lokomoce MeSH
• Proteus mirabilis izolace a purifikace   patogenita   fyziologie   MeSH
• ureasa metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• faktory virulence MeSH
• ureasa MeSH

Cadmium, as a hazardous pollutant commonly present in the living environment, represents an important risk to human health due to its undesirable effects (oxidative stress, changes in activities of many enzymes, interactions with biomolecules including DNA and RNA) and consequent potential risk, making its detection very important. New and unique technological and biotechnological approaches for solving this problems are intensely sought. In this study, we used the commonly occurring potential pathogenic microorganism Staphylococcus aureus for the determination of markers which could be used for sensing of cadmium(II) ions. We were focused on monitoring the effects of different cadmium(II) ion concentrations (0, 1.25, 2.5, 5, 10, 15, 25 and 50 μg mL(-1)) on the growth and energetic metabolism of Staphylococcus aureus. Highly significant changes have been detected in the metabolism of thiol compounds-specifically the protein metallothionein (0.79-26.82 mmol/mg of protein), the enzyme glutathione S-transferase (190-5,827 μmol/min/mg of protein), and sulfhydryl groups (9.6-274.3 μmol cysteine/mg of protein). The ratio of reduced and oxidized glutathione indicated marked oxidative stress. In addition, dramatic changes in urease activity, which is connected with resistance of bacteria, were determined. Further, the effects of cadmium(II) ions on the metabolic pathways of arginine, β-glucosidase, phosphatase, N-acetyl β-d-glucosamine, sucrose, trehalose, mannitol, maltose, lactose, fructose and total proteins were demonstrated. A metabolomic profile of Staphylococcus aureus under cadmium(II) ion treatment conditions was completed seeking data about the possibility of cadmium(II) ion accumulation in cells. The results demonstrate potential in the application of microorganisms as modern biosensor systems based on biological components.

• Klíčová slova
• Brdicka reaction, Staphylococcus aureus, biosensor, cadmium, electrochemistry, high performance liquid chromatography with electrochemical detection, metabolic activity, metabolome, microbiome, spectrophotometry, voltammetry,
• MeSH
• biosenzitivní techniky metody   MeSH
• disacharidy metabolismus   MeSH
• elektrochemické techniky MeSH
• fosfatasy metabolismus   MeSH
• glutathion metabolismus   MeSH
• glutathiondisulfid metabolismus   MeSH
• glutathiontransferasa metabolismus   MeSH
• hydrolasy metabolismus   MeSH
• kadmium analýza   metabolismus   farmakologie   MeSH
• látky znečišťující životní prostředí analýza   metabolismus   farmakologie   MeSH
• metabolismus účinky léků   MeSH
• metalothionein metabolismus   MeSH
• monosacharidy metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• proteiny metabolismus   MeSH
• Staphylococcus aureus cytologie   účinky léků   metabolismus   MeSH
• sulfhydrylové sloučeniny metabolismus   MeSH
• ureasa metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• arginine deiminase MeSH Prohlížeč
• disacharidy MeSH
• fosfatasy MeSH
• glutathion MeSH
• glutathiondisulfid MeSH
• glutathiontransferasa MeSH
• hydrolasy MeSH
• kadmium MeSH
• látky znečišťující životní prostředí MeSH
• metalothionein MeSH
• monosacharidy MeSH
• proteiny MeSH
• sulfhydrylové sloučeniny MeSH
• ureasa MeSH

Molecular cloning, nucleotide sequencing, and characterization of the flaA gene from additional isolates of urease-positive thermophilic Campylobacter (UPTC) were performed. These isolates were obtained from the natural environment in Northern Ireland (n = 9 from mussels) and in England (n = 1 from sea water). All isolates carried the shorter flaA gene, [open reading frames (ORFs), 1,461 to 1,503 base pairs], without any internal termination codons, and did not carry any flaA pseudogenes. The UPTC isolates were well discriminated by the neighbor joining (NJ) phylogenetic tree constructed based on the putative flaA genes ORFs nucleotide sequence information. In addition, the NJ tree constructed based on the flaA-short variable region sequence information discriminated the Campylobacter lari isolates with a similar degree of discrimination power.

• MeSH
• Campylobacter lari klasifikace   genetika   izolace a purifikace   metabolismus   MeSH
• Campylobacter klasifikace   genetika   izolace a purifikace   metabolismus   MeSH
• flagelin chemie   genetika   metabolismus   MeSH
• fylogeneze MeSH
• klonování DNA MeSH
• mlži mikrobiologie   MeSH
• molekulární sekvence - údaje MeSH
• mořská voda mikrobiologie   MeSH
• otevřené čtecí rámce MeSH
• polymerázová řetězová reakce MeSH
• rekombinantní proteiny chemie   genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční analýza DNA metody   MeSH
• sekvenční seřazení MeSH
• ureasa metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Anglie MeSH
• Severní Irsko MeSH
• Názvy látek
• flaA protein, bacteria MeSH Prohlížeč
• flagelin MeSH
• rekombinantní proteiny MeSH
• ureasa MeSH