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ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES
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Diagnostic PCR assay for Microsporum and Trichophyton infections
PublikacjaWe have previously described a highly sensitive 5-hour PCR test for the rapid diagnosis of onychomycosis which detects any dermatophyte and species-identify T. rubrum from patient specimens. We have subsequently developed and evaluated new PCR tests for detection of Trichophyton and Microsporum canis/audouinii infections.58 dermatophyte isolates (21) Microsporum spp. (4 different species), 35 Trichophyton spp (10 different species)...
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A rapid assay of human thyroid peroxidase activity
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A new assay for the simultaneous identification and differentiation of Klebsiella oxytoca strains.
PublikacjaKlebsiella oxytoca is the second most frequently identified species of Klebsiella isolated from hospitalized patients. Klebsiella spp. is difficult to identify using conventional methods and is often misclassified in clinical microbiology laboratories. K. oxytoca is responsible for an increasing number of multi-resistant infections in hospitals because of insufficient detection and identification. In this study, we propose a new...
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A new assay for simultaneous identification and differentiation of Klebsiella oxytoca strains
PublikacjaBacterial strain typing, or identifying bacteria at the strain level, is particularly important for diagnosis, treatment, and epidemiological surveillance of bacterial infections. This is especially the case for bacteria exhibiting high levels of antibiotic resistance or virulence, and those involved in nosocomial or pandemic infections. Strain typing also has applications in studying bacterial population dynamics.
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The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)
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A Colorimetric Microplate Assay for DNA-Binding Activity of His-Tagged MutS Protein
PublikacjaA simple microplate method was designed for rapid testing DNA-binding activity of proteins. The principle of the assay involves binding of tested DNA by his-tagged protein immobilized on a nickel-coated ELISA plate, following colorimetric detection of biotinylated DNA with avidin conjugated to horseradish peroxidase. The method was used to compare DNA mismatch binding activities of MutS proteins from three bacterial species. The...
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Wound healing migration assay
Dane BadawczeThe dataset presents the quantified percentage of wound closure, measured using ImageJ software. The progression of wound width over time was assessed following the treatment of A549, H226, and H460 cells with selected anthraquinones.
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Preliminary antifungal activity assay of selected chlorine-containing derivatives of xanthone and phenoxyethyl amines
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Development and validation of urinary nucleosides and creatinine assay by capillary electrophoresis with solid phase extraction
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Characterization of Selected Jackfruit Germplasm Accessions for Fruit Shape Through Morphological and Marker Based Assay
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Deep eutectic solvents based assay for extraction and determination of zinc in fish and eel samples using FAAS
PublikacjaA new assay based on effective (high recovery) extraction by means of deep eutectic solvents (DESs) was developed for ppb level determination of zinc in fishes and eel samples. Choline chloride and Phenol in a 1:2 M ratio was selected as optimal DES-based extraction solvent. 8-Hydroxy quinoline was used as a chelating agent for zinc ions. The optimized conditions were found at pH value of 8, ligand concentration of 10 mg/L, THF...
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Analysis of the influence of polystyrene microplastics and their derivatives on the DNA of human colon epithelial cells HT29 with the comet assay
PublikacjaWe have been using plastic for almost a century and nowadays a lot of them circulates as pollutants in the environment and still defragmenting to micro and nanoscale. The exposition through the food chain and its precise impact on human health is still not clear. In our study, we tested real food packaging after contact with food products and real thermoinsulation and environmental samples of polystyrene in different model liquids,...
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Optimization and validation of a multiplex assay for microsatellite loci analysis in the field cricket, Gryllus campestris (Orthoptera: Gryllidae)
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Comet assay-based methods for measuring DNA repair in vitro; estimates of inter- and intra-individual variation.
PublikacjaZdolność do naprawy uszkodzeń DNA jest jednym z najważniejszych wyznaczników podatności na choroby nowotworowe. Dlatego też użytecznym staje się możliwość pomiaru stopnia naprawy DNA w próbkach materiału badawczego pobranego od ochotników. Celem pracy było, po pierwsze, opracowanie w warunkach in vitro prostej metody (z wykorzystaniem techniki testu kometowego), w której naprawa DNA odbywa się poprzez wycinanie nukleotydów (NER)....
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Quantification of Radiation-Induced Single-Strand Breaks in Plasmid DNA using a TUNEL/ELISA-Based Assay
PublikacjaWprowadzono nową metodę służącą do oznaczania in vitro pojedynczych pęknięć nici DNA powstałych pod wpływem naświetlań. Porównano wyniki otrzymane z nowej metody z modelem teoretycznym. Porównano wyniki z szeroko używaną metodą elektroforezy żelowej. Pokazano, że poziom pojedynczych pęknięć powstających w niciach DNA pod wpływem promieniowania VUV jest wyższy niż zakładany na podstawie wyników otrzymywanych metodami standardowymi.
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A novel in vitro assay for assessing efficacy and toxicity of antifungals using human leukaemic cells infected with Candida albicans
PublikacjaAims: This study describes a novel in vitro assay that simultaneously determines antifungal efficiency and host cell toxicity using suspensions of human leukaemic cells (HL-60) infected with Candida albicans. Methods and Results: The effect of Candida infection on host cell viability was evaluated by the microscopy of trypan blue-stained cells and lactate dehydrogenase (LDH) activity. The in vitro ‘drug potency assay’ utilized...
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Quality Control of Bacterial Extracellular Vesicles with Total Protein Content Assay, Nanoparticles Tracking Analysis, and Capillary Electrophoresis
PublikacjaExtracellular vesicles (EVs) were isolated from Pectobacterium zantedeschiae culturing media using direct ultracentrifugation (UC), iodixanol cushion ultracentrifugation (ICUC), and iodixanol density gradient ultracentrifugation (IDGUC) techniques. The isolates were characterized with total protein content assay (bicinchoninic acid assay, BCA), nanoparticles tracking analysis (NTA), and capillary electrophoresis (CE). A satisfactory...
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Antiproliferative activity of anthraquinones determined by clonogenic assay
Dane BadawczeThe data contains the colony formation analysis in A549, H460 and H226 cells after exposure to TXT2 and TXT4. DMSO was used as a negative control.
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Toxoplasma gondii: Usefulness of ROP1 recombinant antigen in an immunoglobulin G avidity assay for diagnosis of acute toxoplasmosis in humans
PublikacjaWyniki zawarte w niniejszej pracy pokazują, iż antygen rekombinantowy ROP1 Toxoplasma gondii może być użyteczny w wykrywaniu wczesnej fazy toksoplazmozy z zastosowaniem testu oznaczania awidności przeciwciał klasy IgG. W przeprowadzonych badaniach wykazano, że specyficzne przeciwciała o niskiej awidności były wykrywane w surowicach pacjentów z wczesną fazą choroby, natomiast przeciwciała o wysokiej awidności lub ich brak notowano...
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Toxoplasma gondii: Recombinant GRA5 antigen for detection of immunoglobulin G antibodies using enzyme-linked immunosorbent assay
PublikacjaW publikacji po raz pierwszy oszacowano przydatność diagnostyczną antygenu rekombinantowego GRA5 Toxoplasma gondii do serodiagnostyki toksoplazmozy u ludzi. Do celu wykorzystano pulę surowic pochodzącą od 189 pacjentów chorych na toksoplazmozę wczesną, podostrą i przewlekłą oraz od 31 pacjentów zdrowych , które testowano z wykorzystaniem metody ELISA. Specyficzne przeciwciała anty-GRA5 klasy IgG wykryto w 70,9% surowic seropozytywnych....
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Design, Synthesis, and Enzymatic Evaluation of Novel ZnO Quantum Dot-Based Assay for Detection of Proteinase 3 Activity
PublikacjaHerein, the synthesis and application of functionalized quantum dot-based protease probes is described. Such probes are composed of nontoxic ZnO nanocrystals decorated by amino groups followed by linker and labeled peptide attachment. Spherical NH2-terminated ZnO quantum dots (QDs) with the average size ranging from 4 to 8 nm and strong emission centered at 530 nm were prepared using the sol−gel method. The fluorescence of ZnO...
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Deep eutectic solvent (DES) with silver nanoparticles (Ag-NPs) based assay for analysis of lead (II) in edible oils
PublikacjaThis paper presents an application of silver nanoparticles impregnated by Deep Eutectic Solvents (DES) as ultrasonication aided microextraction system for lead (II) determination in edible oils. The paper presents a systematic optimization of method parameters and examples of its application for analysis of real samples. Maximum recovery for lead (II) extraction was obtained for choline chloride and phenol with a 1:2 molar ratio....
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Monitoring T-Cell Responses in Translational Studies: Optimization of Dye-Based Proliferation Assay for Evaluation of Antigen-Specific Responses
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Physicochemical profile of Os (III) complexes with pyrazine derivatives: From solution behavior to DNA binding studies and biological assay
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A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification
PublikacjaCandida sp. have been responsible for an increasing number of infections, especially in patients with immunodeficiency. Species specific differentiation of the Candida sp. is difficult in routine diagnosis. This identification can have a highly significant association in therapy and prophylaxis. This work has shown a new application of terminal restriction fragment length polymorphism (t-RFLP) method in molecular identification...
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Assay of Cystathionine-γ-synthase (CaStr2p) activity determination.
Dane BadawczeThe study aimed to obtain conditions optimal for the activity of Candida albicans cystathionine-γ-synthase (CaStr2p) determination. The selection of appropriate reaction conditions included the identification of the reaction buffer and its pH, substrate, and enzyme concentrations. The activity of Str2p was measured by the detection of decrease of the...
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Corrigendum: Monitoring T-Cell Responses in Translational Studies: Optimization of Dye-Based Proliferation Assay for Evaluation of Antigen-Specific Responses
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Introduction of a multiplex amplicon sequencing assay to quantify DNA methylation in target cytosine markers underlying four selected epigenetic clocks
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Multiplex PCR assay for the simultaneous identification of race specific and non-specific leaf resistance genes in wheat (Triticum aestivum L.)
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Serodiagnosis of Toxoplasma gondii infection in farm animals (horses, swine, and sheep) by enzyme-linked immunosorbent assay using chimeric antigens
PublikacjaToxoplasma gondii infects all warm-blooded animals including humans, causing serious public health problems and great economic loss in the animal husbandry. Commonly used serological tests for diagnosis of toxoplasmosis involve preparation of whole Toxoplasma lysate antigen (TLA) from tachyzoites. The production of this antigen is associated with high costs and lengthy preparation and the possibility of staff infection. There are...
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Activity assay of O-Acetyl-L-homoserine sulfhydrylase (CaMet15p).
Dane BadawczeThe study aimed to obtain conditions optimal for the activity of O-acetyl-L-homoserine sulfhydrylase (CaMet15p) and a method suitable for its measurement. The selection of appropriate reaction conditions included the identification of the reaction buffer and its pH, substrate and enzyme concentrations. The activity of CaMet15p was measured by the detection...
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Toxoplasma gondii: Enzyme-Linked Immunosorbent Assay using different fragments of recombinant microneme protein 1 (MIC1) for detection of immunoglobulin G antibodies
PublikacjaTrzy rekombinantowe fragmenty białka antygenowego MIC1 Toxoplasma gondii (r-MIC1ex2, r-MIC1ex34 i r-MIC1) zostały wyprodukowane jako białka fuzyjne (zawierające dwie domeny oligohistydynowe na N- i C-końcu) w komórkach bakteryjnych Escherichia coli. Homogenne preparaty antygenów rekombinantowych otrzymane z zastosowaniem jednoetapowego oczyszczania metodą chromatografii metalopowinowactwa, wykorzystano następnie do immunoidentyfikacji...
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Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: Multivariate data analysis of multisine impedimetric response
PublikacjaSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the cells through the binding of spike protein to the host cell surface-expressing angiotensin-converting enzyme 2 (ACE2) or by endocytosis mediated by extracellular matrix metalloproteinase inducer (CD147). We present extended statistical studies of the multisine dynamic electrochemical impedance spectroscopy (DEIS) revealing interactions between Spike RBD and...
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Hybrid metal and non-metal activation of Oxone by magnetite nanostructures co-immobilized with nano-carbon black to degrade tetracycline: Fenton and electrochemical enhancement with bio-assay
PublikacjaElectrochemically synthesized magnetite nanostructures (ESMNPs) as a metal activator and nano-carbon black (NCB) as a non-metal activator were co-immobilized by alginate natural polymer to activate Oxone for the degradation of tetracycline (TC) antibiotic. The formation of sulfate radical was indirectly confirmed during the Oxone/ESMNPs/NCB/alginate process via the addition of scavenging compounds. This study revealed the high...
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Enzymatic cleavage of specific dipeptide conjugated with ferrocene as a flexible ultra-sensitive and fast voltammetric assay of matrix metalloproteinase-9 considered a prognostic cancer biomarker in plasma samples
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The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants
PublikacjaThis study presents an evaluation of seventeen newly produced recombinant trivalent chimeric proteins (containing the same immunodominant fragment of SAG1 and SAG2 of Toxoplasma gondii antigens, and an additional immunodominant fragment of one of the parasite antigens, such as AMA1, GRA1, GRA2, GRA5, GRA6, GRA7, GRA9, LDH2, MAG1, MIC1, MIC3, P35, and ROP1) as a potential alternative to the whole-cell tachyzoite lysate (TLA) used...
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Association between plasma proteome profiles analysed by mass spectrometry, a lymphocyte-based DNA-break repair assay and radiotherapy-induced acute mucosal reaction in head and neck cancer patients
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The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants
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Study of the ability of TXT-compounds to promote telomeric G4 DNA thermal stabilization using FRET melting assay
Dane BadawczeThe data set contains a DNA melting screen obtained from the FRET melting assay. Thermal stabilities of G-quadruplex structures were analysed in the presence and in absence of ligands. TMPyP4 and MTX were used as references. Measurements were carried out in triplicate using a LightCycler 480 thermocycler (Roche Life Science) with a filter setting of...
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Antitumor Activity of Triazine Mimic Antibiotics for DNA-binding Implications (Impressive Activity in Vitro Against a Variety of Tumor Types in the NCI-60 Screen): NSC 710607 To Fight HCT-116 Human Colon Carcinoma Cell Lines in Vivo Using the Hollow Fiber Assay and Xenograft Mouse Models
PublikacjaPurpose Successful clinical applications of DNA-directed selective cytotoxic agents disrupt the vital replication/transcription processes and ultimately lead to cancer cell death. This study aimed to examine the growth screen of two lead triazine compounds in a number of cell lines and xenografts and to develop anticancer agents with noncovalent binding affinity bringing fewer side effects. Methods The NCI initial hollow...
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Future of Protease Activity Assays
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Methods of assaying volatile oxygenated organic compounds in effluent samples by gas chromatography—A review
PublikacjaThe paper is a review of the procedures for the determination of volatile and semivolatile oxygenated organic compounds (O-VOCs) in effluent samples by gas chromatography. Current trends and outlook for individual steps of the procedure for the determination of O-VOCs in effluents are discussed. The available sample preparation techniques and their limitations are described along with GC capillary columns used for O-VOCs separation...
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Some analytical assays for the determination of bioactivity of exotic fruits
PublikacjaPrzedmiotem badań były próbki owoców egzotycznych (mango, durian, gruszka adwokacka) zebrane w 2008 roku w Tajlandii i w Izraelu (po etapie liofilizacji). W próbkach oznaczano zawartość:- białek, - tłuszczów,- węglowodanów,- flawonoidów,- tanin- polifenoli stosując zarówno zaawansowane techniki instrumentalne jak i proste testy spektrofotometryczne (FRAP, ABTS, DPPH, CUPRAC). Uzyskana wieloparametrowa baza danych pomiarowych została...
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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).
PublikacjaNo Abstract
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INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies
PublikacjaIn vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant pancreatic enzymes such as proteases, amylase and lipase. Rabbit gastric extracts (RGE) have been recently introduced to provide gastric lipase in addition to pepsin. Before preparing simulated gastric and pancreatic extracts with...
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Analytical aspects of model compounds toxicity assessment using MTT and Microtox bioanalytica assays
PublikacjaMore and more increased intensity of anthropopressure processes can be observed, among other things, in the release of great quantities of synthetic substances into the environment, including the Endocrine Disrupting Compounds. The effects of the above-mentioned human impact on the environment not only can be noticed in the homeostasis disturbance in ecosystems, but they are also visible with respect to human. The high incidence...
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PCR and real-time PCR assays to detect fungi of Alternaria alternata species
PublikacjaFungi of the Alternaria genus are mostly associated with allergic diseases. However, with a growing number of immunocompromised patients, these fungi, with A. alternata being the most prevalent one, are increasingly recognized as etiological agents of infections (phaeohyphomycoses) in humans. Nowadays, identification of Alternaria spp. requires their pure culture and is solely based on morphological criteria. Clinically, Alternaria...
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Rapid Assays for Specific Detection of Fungi of Scopulariopsis and Microascus Genera and Scopulariopsis brevicaulis Species
PublikacjaPurpose Fungi of Scopulariopsis and Microascus genera cause a wide range of infections, with S. brevicaulis being the most prevalent aetiological agent of mould onychomycosis. Proper identification of these pathogens requires sporulating culture, which considerably delays the diagnosis. So far, sequencing of rDNA regions of clinical isolates has produced ambiguous results due to the lack of reference sequences in publicly available...
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PCR-RFLP assays for species-specific identification of fungi belonging to Scopulariopsis and related genera
PublikacjaFungi of the Scopulariopsis genus, commonly found in the environment, are opportunistic pathogens that can cause various types of human infections. So far, no efficient molecular method has been developed for species differentiation among Scopulariopsis and related genera. In order to advance this field, we have evaluated performance of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assays, based...