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Detection of Toxoplasma gondii in meat - available techniques
PublicationToxoplasma gondii, which belongs to the phylum Apicomplexa, is an absolute intercellular parasite that infects a wide range of hosts, including warm-blooded animals and humans. An infection with this parasite causes toxoplasmosis, which in most cases remains latent. However, this disease is a major threat to immunocompromised people and pregnant women. In extreme cases, it can damage nervous system or case fatal abortion. Infected...
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Grzyby środowiskowe jako biologiczne wskaźniki jakości powietrza w budynkach
PublicationAnaliza mikrobiologiczna powietrza ulega ciągłym usprawnieniom począwszy od lat czterdziestych XX wieku. Dzięki temu wykazano, iż grzyby środowiskowe (ich elementy lub substancje przez nie produkowane) mogą wpływać na stan powietrza w budynkach. Wysunięcie tych wniosków zeszło się w czasie z odkryciem Zespołu Chorego Budynku. Podczas gdy w normalnym powietrzu w budynkach dominuje rodzaj Cladosporium, w chorych budynkach znajduje...
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Problem reakcji krzyżowych w immunodiagnostyce na przykładzie neosporozy i toksoplazmozy
PublicationW niniejszej pracy przedstawiono problem reakcji krzyżowych występujących w immunodiagnostyce chorób zakaźnych. Zagadnienie opisano szerzej na przykładzie dwóch blisko spokrewnionych pasożytów należących do typu Apicomplexa - Neospora caninum i Toxoplasma gondii. Zaprezentowano także, sposoby ulepszania testów wykorzystywanych w diagnostyce neosporozy i toksoplazmozy u zwierząt.
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Genetic predisposition to inflammatory bowel disease
PublicationInflammatory bowel disease (IBD) is a chronic, incurable inflammatory disease of the digestive system. The two main disease entities included in the IBD are ulcerative colitis and Crohn's disease. According to epidemiological studies there are more and more new cases every year. In especially among the youngest patients with symptoms of malnutrition and growth inhibition to land up in hospitalwith cancer suspected. The purpose...
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Sick building syndrome due to molds
PublicationThe term Sick Building Syndrome (SBS) is a combination of ailments occurring in a specific building. Building occupants complain of symptoms such as headaches and dizziness, fainting, nausea, irritation of the mucosal membranes, skin irritation and other nonspecific hypersensitivity reactions.One of the SBS causes can be molds, which are ubiquitous in the air (over 400 species). It is estimated that 30% of the health problems (related...
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Identification of Antigen 43 in uropathogenic Escherichia coli IH11128 responsible for persistence in the urinary tract
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Production of Toxoplasma gondii MAG1 recombinant antigen in Leishmania tarentolae expression system
PublicationThe results of the work associated with the production of the Toxoplasma gondii matrix antigen 1 (MAG1) in the eukaryotic expression system of Leishmania tarentolae are shown.
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Postępy w zakresie analizy powietrza mającej na celu identyfikację grzybów środowiskowych
PublicationPowietrze jest środowiskiem znacznie bliższym człowiekowi niż woda i gleba. Zatem jakość powietrza, szczególnie w pomieszczeniach, w których człowiek spędza nawet 80-95% czasu dnia jest jednym z najistotniejszych czynników wpływających na ogólną jakość życia ludzi. Skażenie powietrza w pomieszczeniach może wywoływać problemy zdrowotne. Jednym ze składników zanieczyszczeń powietrza są grzyby środowiskowe, które mogą wpływać na zdrowie...
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Pilicides effectively hinder chaperone-usher interactions in all Gram-negative bacteria
PublicationThe chaperone-usher pathway of adhesive structures biogenesis is highly conserved in all Gram-negative bacteria. Pili are polymers of thousands protein subunits that possess conserved immunoglobuline-like structure denoted by lack of the seventh G strand. The effect of this structural defect is a hydrophobic acceptor cleft. The folding of protein subunits is strictly dependent on the action of specific periplasmic chaperone protein...
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Reakcje krzyżowe w immunodiagnostyce
PublicationZaprezentowano problem reakcji krzyżowych w immunodiagnostyce.
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Hyperproteic diet modifies microbiota composition and associated metabolites in rats
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Nieswoiste zapalenie jelit
PublicationNieswoiste zapalenie jelit jest przewlekłą, nieuleczalną choroba zapalną układu pokarmowego. W niniejszej pracy przedstawiono dane epidemiologiczne, możliwe przyczyny wystąpienia choroby oraz opisano dwie główne jednostki chorobowe zaliczane do nieswoistego zapalenia jelit takie jak wrzodziejące zapalenie jelita grubego oraz choroba Leśniowskiego-Crohn'a.
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Molecular detection of Candida krusei
PublicationThe species identification of fungi belonging to genus Candida is an important issue as this genus becomes the emerging problem of nosocomial infections. As Candida krusei presents intrinsicresistance to the fluconazole that is the drug of the first choice in case of invasive candidiasis the PCR identifying the DNA of C. krusei was elaborated. The analytical sensitivity of the assay on spiked blood samples was estimated at 3-5...
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Novel primosomal protein B from thermophilic bacterium Thermoanerobacter tengcongensis
PublicationWe report the identification and characterization of theprimosomal protein B (PriB) from thermophilic bacteriumThermoanerobacter tengcongensis (TtePriB). It is the largestknown bacterial PriB protein consisting 216 amino acidresidues with a calculated molecular mass of 25 kDa. Surprisingly,it is functional as monomer containing two single-stranded DNA binding domain (OB-fold) and it is thecompletely new kind structure of SSB protein....
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Podłoże nieswoistego zapalenia jelit
PublicationZaprezentowano możliwe czynniki ryzyka wystąpienia nieswoistego zapalenia jelit.
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Covalent DNA modification by products of myrosinase catalysed glucosinolate degradation in cell-free system
PublicationThe bioactive phytochemicals found in Brassica vegetables belonging to glucosinolates (GLS) and especially the products of their degradation isothiocyanates (ITC) and indoles are regarded as the most promising cancer chemopreventive compounds. These secondary metabolites constitute defence system repelling or preventing the development of agrophages attacking brassica plants. The antibiological properties of these compounds suggest...
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Novel DNA-binding protein from Nanoarchaeum equitans Kin4-M binds all kinds of nucleic acids
PublicationNanoarchaeum equitans is the only known representative of Archaea phylum Nanoarchaeota and stands out as one of the tiniest known living organism. What is more it has smallest genome, which is only 490.885 base pairs long. It is also one of the most compact genomes. According to predictions about 95% of the DNA encodes proteins or stable RNA. Nanoarchaeum equitans lacks genes for most vital metabolic pathways including lipid, cofactor,...
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Real-Time PCR: molecular technique of many applications
PublicationReal-Time PCR is a sensitive DNA amplification technique initially applied in genetics and molecular biology. It enables in vivo copying of the selected DNA fragment (flanked by two primers) by the thermostable polymerase (in the presence of magnesium ions and deoxynucleotide triphosphates) and simultaneous measurement of the fluorescence. For one or more specific sequences in a DNA sample, real-time PCR enables both detection...
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Importance of biofilm formation by uropathogenic E. coli Dr+ strains in urinary tract infections
PublicationImportance of biofilm formation by uropathogenic E. coli Dr+ strains in urinary tract infections
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Awidność przeciwciał IgG jako ważny test diagnostyczny w rozpoznawaniu aktywnej toksoplazmozy - stan obecny i nowe możliwości.
PublicationW artykule opisano problem diagnostyki toksoplazmozy ze szczególnym uwzględnieniem metody awidności przeciwciał klasy IgG. Test oznaczania awidności przeciwciał IgG swoistych dla T. gondii jest użytecznym wskaźnikiem serologicznym toksoplazmozy, który w wielu przypadkach na podstawie pojedynczego badania surowicy pacjenta pozwala potwierdzić lub wykluczyć aktywną postać choroby. W pierwotnej, aktywnej toksoplazmozie przeciwciała...
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Peptidoglycan hydrolases - potential weapons against Staphylococcus aureus
PublicationBacteria of the genus Staphylococcus are common pathogens responsible for a broad spectrum of human and animal infections and belong to most important etiological factors causing food poisoning. Because of rapid increase in prevalence of isolation of staphylococci resistant to many antibiotics, there is an urgent need for development of new alternative chemotherapeutics. A number of studies have recently demonstrated the strong...
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Diagnostyka zakażeń Candida spp. oparta o amplifikację DNA techniką PCR
PublicationZakażenia wywoływane przez oportunistyczne grzyby z rodzaju Candida są poważnym problemem klinicznym. Niezbędne jest poszukiwanie nowych szybkich, czułych i pewnych metod do identyfikacji gatunkowej tych drobnoustrojów. W artykule przedstawiono opisane w literaturze metody diagnostyki molekularnej drożdżaków z rodzaju Candida oparte o amplifikację DNA techniką PCR.
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Investigation of interaction between xanthene dye rhodamine B and metagenomic-derivated methylthioadenosine phosphorylase RSFP
PublicationIn the previous study we found that the expression of the metagenomic - derived methylthioadenosine phosphorylase gene (rsfp) in E. coli cells and the presence of rhodamine B in the growth medium were crucial for the pink fluorescence of one E. coli colony from Antarctic soil metagenomic library (Cieśliński et al., 2009). In this study, rsfp gene expression was induced by an arabinose promoter in an LMG194 E. coli strain and the...
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Dimeric structure of the N-terminal domain of PriB protein from Thermoanaerobacter tengcongensis solved ab initio
PublicationPriB is one of the components of the bacterial primosome, which catalyzes the reactivation of stalled replication forks at sites of DNA damage. The N-terminal domain of the PriB protein from the thermophilic bacterium Thermoanaerobacter tengcongensis (TtePriB) was expressed and its crystal structure was solved at the atomic resolution of 1.09 Å by direct methods. The protein chain, which encompasses the first 104 residues of the...
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Development of Safirinium dyes for new applications: fluorescent staining of bacteria, human kidney cells, and the horny layer of the epidermis
PublicationLow-molecular synthetic fluorophores are convenient tools in bioimaging applications. Several derivatives of Safirinium dyes as well as their reactive N-hydroxysuccinimide (NHS) esters bearing diverse substituents were synthesized and evaluated experimentally in terms of their lipophilicity by means of reverse-phase and immobilized artificial membrane high-performance liquid chromatography. Subsequently, the selected compounds...
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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
PublicationWe 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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PRZYDATNOŚĆ DIAGNOSTYCZNA REAKCJI PCR W ROZPOZNAWANIU BORELIOZY
PublicationBorelioza jest wieloukładową chorobą wywoływaną przez bakterie zaliczane do kompleksu Borrelia burgdorferi sensu lato. Wektorem przenoszącym infekcję są kleszcze z rodzaju Ixodes, które zakażają kolejnych żywicieli krętka podczas spożywania ich krwi. Zróżnicowany przebieg boreliozy uniemożliwia jej rozpoznanie na podstawie objawów klinicznych. W związku z tym diagnostyka choroby z Lyme opiera się głównie na metodach laboratoryjnych,...
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Fibrillar aggregates in powdered milk
PublicationThis research paper addresses the hypothesis that powdered milk may contain amyloid fibrils. Amyloids are fibrillar aggregates of proteins. Up to this time, research on the presence of amyloids in food products are scarce. To check the hypothesis we performed thioflavin T fluorescence assay, X-ray powder diffraction, atomic force microscopy and fluorescence microscopy imaging. Our preliminary results show that commercially available...
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Management of ground tire rubber waste by incorporation into polyurethane-based composite foams
PublicationRapid economic growth implicated the developing multiple industry sectors, including the automotive branch, increasing waste generation since recycling and utilization methods have not been established simultaneously. A very severe threat is the generation of enormous amounts of post-consumer tires considered burdensome waste, e.g., due to the substantial emissions of volatile organic compounds (VOCs). Therefore, it is essential...
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Detection of Toxoplasma gondii Infection in Small Ruminants: Old Problems, and Current Solutions
PublicationToxoplasmosis is a parasitic zoonosis of veterinary importance, with implications for public health. Toxoplasma gondii infection causes abortion or congenital disease in small ruminants. Moreover, the consumption of infected meat, cured meat products, or unpasteurized milk and dairy products can facilitate zoonotic transmission. Serological studies conducted in various European countries have shown the high seroprevalence of specific...
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ANTYGENY REKOMBINANTOWE W DIAGNOSTYCE SEROLOGICZNEJ BORELIOZY
PublicationBorelioza jest najczęstszą chorobą odkleszczową dotykającą mieszkańców półkuli północnej. Chorobę tę wywołują bakterie zakwalifikowane do grupy Borrelia burgdorferi sensu lato. Współcześnie podstawą diagnostyki laboratoryjnej boreliozy jest dwustopniowe badanie serologiczne. Pierwszym etapem jest test immunoenzymatyczny (ELISA), jeżeli wynik badania jest dodatni lub wątpliwy jako test potwierdzający stosuje się technikę Western...
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Post-Effort Changes in Autophagy- and Inflammation-Related Gene Expression in White Blood Cells of Healthy Young Men
PublicationAcute, strenuous physical exertion requiring high levels of energy production induces the production of reactive oxygen species and metabolic disturbances that can damage the mitochondria. Thus, selective autophagic elimination of defective mitochondria may improve resistance to oxidative stress and potentially to inflammation. The main goal of this study was to evaluate the impacts of intense effort on changes in the expression...
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Evaluating the antibacterial activity of muramyl dipeptide derivatives, retro-tuftsin derivatives, and anthraquinone oligopeptides against a range of pathogenic bacteria
PublicationSearch for new and efficient antibiotic is crucial because of microbial drug resistance and problems with side effects of the administered medication. In this study, we evaluate the in vitro microbiological activity of muramyl dipeptide derivatives, retro-tuftsin derivatives (i.e., tuftsin with reversed amino acid sequences), and combinations of retro-tuftsin derivatives with substituted anthraquinones. The potency of the investigated...
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Activity of isavuconazole and other triazole derivatives against clinical isolates of Aspergillus fumigatus
PublicationAspergillus fumigatus is the most frequent pathogen of the genus Asperillus, which is highly susceptible to triazole derivatives, especially to isavuconazole and voriconazole. Many countries face a growing problem of infections due to A. fumigatus showing acquired resistance to one or several triazoles. In medical centres, monitoring the susceptibility of isolated Aspergillus spp. is recommended. The aim of this study was to collect...
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A cryptic ribosome binding site, false signals in reporter systems and avoidance of protein translation chaos
PublicationThe expression of reporter gene may be induced by activation of cryptic signalling sequences, as we found while constructing the mutS-lacZ fusion gene. We cloned the Escherichia coli lacZ gene encoding beta-galactosidase into a plasmid vector carrying the Thermus thermophilus mutS gene. The clones expected to produce beta-galactosidase as the C-terminal fusion were selected for the complementation of beta-galactosidase activity...
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Total impact of oxidative stress genes on cardiovascular events—a 7-year follow-up study
PublicationCardiovascular (CV) events are the number one cause of lifetime disability and deaths worldwide. It is well known that traditional risk factors do not fully correlate with clinical outcomes; therefore, searching for other markers that would explain CV events occurrence seems essential. Of importance, one of the main factors at the origin of CV events is oxidative stress, causing inflammation and atherosclerotic plaque instability....
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Optical method supported by machine learning for urinary tract infection detection and urosepsis risk assessment
PublicationThe study presents an optical method supported by machine learning for discriminating urinary tract infections from an infection capable of causing urosepsis. The method comprises spectra of spectroscopy measurement of artificial urine samples with bacteria from solid cultures of clinical E. coli strains. To provide a reliable classification of results assistance of 27 algorithms was tested. We proved that is possible to obtain...
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Expression of goose parvovirus whole VP3 protein and its epitopes in Escherichia coli cells
PublicationThe aim of this study was the expression of goose parvovirus capsid protein (VP3) and its epitopes in Escherichia coli cells. Expression of the whole VP3 protein provided an insufficient amount of protein. In contrast, the expression of two VP3 epitopes (VP3ep4, VP3ep6) in E. coli, resulted in very high expression levels. This may suggest that smaller parts of the GPV antigenic determinants are more efficiently expressed than the...
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An examination of an Antarctic soil metagenomic-derivate putative methylthioadenosine phosphorylase gene as a novel reporter gene for promoter trapping
PublicationThe rsfp gene was discovered during examination ofan Antarctic soil-derived metagenome library and encodesa bacterial methylthioadenosine phosphorylase (GenBank accession number GQ202582). In a previous study, we found that E. coli colonies expressing the rsfp gene reveal strong pink fluorescence after exposure to UV light when grown on culture medium supplemented with rhodamine B (Cieśliński et al., 2009). In contrast to that,...
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Regenerative Drug Discovery Using Ear Pinna Punch Wound Model in Mice
PublicationThe ear pinna is a complex tissue consisting of the dermis, cartilage, muscles, vessels, and nerves. Ear pinna healing is a model of regeneration in mammals. In some mammals, including rabbits, punch wounds in the ear pinna close spontaneously; in common-use laboratory mice, they remain for life. Agents inducing ear pinna healing are potential regenerative drugs. We tested the effects of selected bioactive agents on 2 mm ear pinna...
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Impact of temperature on optical sensing in biology based on investigation of SARS‐CoV ‐2
PublicationIn this paper, we present an investigation of the influence of the temperature on the sensing of biological samples. We used biofunctionalized microsphere-based fiber-optic sensor to detect immunoglobulin G attached to the sensor head at temperatures relevant in biological research: 5°C, 25°C, and 55°C. The construction of the sensor allowed us to perform measurements in the small amount of solution. The results of our experiment...
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Identification of evolutionary conserved DNA sequence and corresponding S21 ribosomal protein region for diagnostic purposes of all Borrelia spirochetes
PublicationIt is still under investigation, whether all Borrelia sp. causing Lyme borreliosis and other diseases are already identified and properly classified as human pathogens. For this reason, it is of great importance to develop a diagnostic ELISA test that detects all Borrelia sp. The aim of this study was to identify conserved DNA and protein regions present in all currently known Borrelia sp. In experimental studies 31 available Borrelia...
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Asymptomatic Bacteriuria in Kidney Transplant Recipients—A Narrative Review
PublicationUrinary tract infections (UTIs) are the most prevalent complications in kidney transplant (KTx) recipients. The most frequent finding in this group of patients is asymptomatic bacteriuria (ASB). Here, we provide an overview of the available evidence regarding ASB in KTx recipients, including its etiopathology, clinical impact and management. There is a growing body of evidence from clinical trials that screening for and treating...
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Searching for the primary metabolic alterations of polycystic ovary syndrome by application of the untargeted metabolomics approach
PublicationDespite a large number of studies, the pathogenesis of polycystic ovary syndrome (PCOS) still remains unexplained. In light of ambiguous observations reported in metabolomics, there is a need to carry out studies focusing on confirming the discriminating power of the proposed metabolomics biomarkers. Our research aimed to perform a validation study of metabolites detected in our previous study from serum samples, on the new set...
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PCR-RFLP assays for species-specific identification of fungi belonging to Scopulariopsis and related genera
PublicationFungi 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...
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May Staphylococcus lugdunensis Be an Etiological Factor of Chronic Maxillary Sinuses Infection?
PublicationStaphylococcus lugdunensis is an opportunistic pathogen found in the healthy human skin microbiome bacterial community that is able to cause infections of diverse localization, manifestation, and course, including laryngological infections, such as necrotizing sinusitis. Chronic maxillary sinusitis is a disease present in up to one third of European and American populations, and its etiology is not fully described. Within this...
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A New Expression System Based on Psychrotolerant Debaryomyces macquariensis Yeast and Its Application to the Production of Cold-Active β-D-Galactosidase from Paracoccus sp. 32d
PublicationYeasts provide attractive host/vector systems for heterologous gene expression. The currently used yeast-based expression platforms include mesophilic and thermotolerant species. A eukaryotic expression system working at low temperatures could be particularly useful for the production of thermolabile proteins and proteins that tend to form insoluble aggregates. For this purpose, an expression system based on an Antarctic psychrotolerant...
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Cloning, expression in Komagataella phaffii, and biochemical characterization of recombinant sequence variants of Pseudomonas sp. S9 GDSL-esterase
PublicationTwo recombinant Komagataella phaffii (formerly Pichia pastoris) yeast strains for production of two sequential variants of EstS9 esterase from psychrotolerant bacterium Pseudomonas sp. S9, i.e. αEstS9N (a two-domain enzyme consisting of a catalytic domain and an autotransporter domain) and αEstS9Δ (a single-domain esterase) were constructed. However, only one of recombinant K. phaffii strains, namely Komagataella phaffii X-33/pPICZαestS9Δ,...
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Post-match recovery profile of leukocyte cell subsets among professional soccer players
PublicationThis study assessed the impact of cumulative match time on the distribution of CD45+ cell subtests in the capillary blood of professional soccer players. Twenty-two males (aged 18–30 years) took part in the 36-week study. Participants playing up to 540 in cumulative match time and less than 30 min in each single match during the observation period formed the control group. White blood cell (WBC) phenotyping and creatine kinase...
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Evaluation of long‑term immunity and protection against T. gondii after immunization with multivalent recombinant chimeric T. gondii proteins
PublicationToxoplasmosis caused by the opportunistic, cosmopolitan protozoan Toxoplasma gondii is one of the most common parasitoses in the world. Although it may prove dangerous or even fatal for immunocompromised individuals, immunoprophylaxis for humans is still nonexistent. Thus, the aim of the current work was to assess the ability of two immunogenic recombinant chimeric T. gondii proteins, SAG2-GRA1-ROP1 (SGR) and SAG1-MIC1-MAG1-GRA2...
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A new division of bacterial UvrA homologues
PublicationThe UvrA protein is a DNA-binding and damage-recognition enzyme which participates in the prokaryotic type nucleotide excision repair (NER) pathway. It has recently been noted that some bacterial genomes comprise additional uvrA genes which encode five distinct types of UvrA homologue. We investigated the sequences of over 2400 bacterial genomes and found 130 examples of bacteria containing uvrA 2 genes. The sequence analyses conducted...
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Patomechanizm zakażeń dróg moczowych wywoływanych przez uropatogenne szczepy E. coli
PublicationZakażenia układu moczowego (ZUM) to jedne z najczęstszych i najbardziej powszechnych infekcji bakteryjnych, dotykających każdego roku nawet 150 milionów ludzi na świecie. Problem z tego typu infekcjami wynika z przewlekłości choroby, nawrotów i wzrastającej lekooporności powodujących je patogenów. Uropatogenne szczepy E. coli (UPEC) są głównym czynnikiem przyczynowym ZUM. Bakterie tej grupy zawierają wiele czynników adhezyjnych...
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First report on echinocandin resistant Polish Candida isolates
PublicationPurpose: Candida spp. are ranked as one of the four major causative agents of fungal infections. The number of infections caused by Candida species resistant to fluconazole, which is applied as the first line drug in candidiasis treatment, increases every year. In such cases the application of echinocandin is necessary. Echinocandin susceptibility testing has become a routine laboratory practice in many countries due to the increasing...
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Metabolic activity of tree saps of different origin towards cultured human cells in the light of grade correspondence analysis and multiple regression modeling
PublicationTree saps are nourishing biological media commonly used for beverage and syrup production. Although the nutritional aspect of tree saps is widely acknowledged, the exact relationship between the sap composition, origin, and effect on the metabolic rate of human cells is still elusive. Thus, we collected saps from seven different tree species and conducted composition-activity analysis. Saps from trees of Betulaceae, but not from...
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A novel method of Mycobacterium tuberculosis complex strain differentiation using polymorphic GC-rich gene sequences.
PublicationTuberculosis is one of the leading infectious diseases. In this work, a new genotyping method of Mycobacterium tuberculosis (Mtb) complex strain is presented. 27 Mtb genomes were analyzed for the presence of length polymorphism within polymorphic GC-rich gene sequences. Four genes, Rv3345c, Rv3507, Rv0747 and Rv3511, showing variation in length depending on the Mtb strain were selected for designing primer sequences flanking variable...
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Relation of the polymorphism of cyp51A sequence and the susceptibility of Aspergillus fumigatus isolates to triazoles determined by commercial gradient test (Etest) and by reference methods
PublicationThe aim of this study was to evaluate the accuracy of commercial gradient test (Etest) in the detection of triazole resistant Aspergillus fumigatus isolates using reference microdilution methods and the analysis of sequences of the cyp 51A gene. The study was performed on twenty clinical isolates which were identified as Aspergillus fumigatus based on the DNA sequences of the ITS1-2 fragment of ribosomal DNA and the β-tubulin gene,...
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A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification
PublicationCandida 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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In vitro affinity of Deinococcus radiodurans MutS towards mismatched DNA exceeds that of its orthologues from Escherichia coli and Thermus thermophilus
PublicationThe mismatch binding protein MutS is responsible for the recognition of mispaired and unpaired bases, which is the initial step in DNA repair. Among the MutS proteins most extensively studied in vitro are those derived from Thermus thermophilus, Thermus aquaticus and Escherichia coli. Here, we present the first report on the in vitro examination of DNA mismatch binding activity of MutS protein from Deinococcus radiodurans and confront...
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Analysis of the unique structural and physicochemical properties of the DraD/AfaD invasin in the context of its belonging to the family of chaperone/usher type fimbrial subunits
PublicationInwazyna DraD kodowana przez operon dra posiada strukturę charakterystyczną dla białek podjednostkowych fimbrii typu chaperone-usher. Fold-Ig białka DraD posiada dwie charakterystyczne cechy wyróżniające je z pośród rodziny fimbrialnych białek podjednostkowych: 1) zaburzenie struktury beta-beczki w rejonie szczeliny akceptorowej objawiające się układem wiązań wodorowych głównego łańcucha polipeptydowego oraz 2) nietypowo usytuowany...
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X-ray and UV Radiation Damage of dsDNA/Protein Complexes
PublicationRadiation and photodynamic therapies are used for cancer treatment by targeting DNA. However, efficiency is limited due to physico-chemical processes and the insensitivity of native nucleobases to damage. Thus, incorporation of radio- and photosensitizers into these therapies should increase both efficacy and the yield of DNA damage. To date, studies of sensitization processes have been performed on simple model systems, e.g.,...
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Generation and Characterization of a DNA-GCN4 Oligonucleotide-Peptide Conjugate: The Impact DNA/Protein Interactions on the Sensitization of DNA
PublicationRadiotherapy, the most common therapy for the treatment of solid tumors, exerts its effects by inducing DNA damage. To fully understand the extent and nature of this damage, DNA models that mimic the in vivo situation should be utilized. In a cellular context, genomic DNA constantly interacts with proteins and these interactions could influence both the primary radical processes (triggered by ionizing radiation) and secondary reactions,...
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Association of Genes Related to Oxidative Stress with the Extent of Coronary Atherosclerosis
PublicationOxidative stress is believed to play a critical role in atherosclerosis initiation and progression. In line with this, in a group of 1099 subjects, we determined eight single nucleotide polymorphisms (SNPs) related to oxidative stress (PON1 c.575A>G, MPO c.463G>A, SOD2 c.47T>C, GCLM c.590C>T, NOS3 c.894G>T, NOS3 c.786T>C, CYBA c.214C>T, and CYBA c.932A>G) and assessed the extent of atherosclerosis in coronary arteries based on...
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Plasma free amino acid profiling as metabolomic diagnostic and prognostic biomarker in paediatric cancer patients: a follow‑up study
PublicationAmino acids (AAs) play a crucial role in cancer cell metabolism. Levels of 22 plasma AAs at the time of diagnosis and after treatment were established among 39 pediatric cancer patients and 33 healthy children. Glutamic acid levels decreased and tryptophan levels increased during treatment. Cancer patients presented significantly lower levels of glutamine and leucine post-treatment while levels of 12 other AAs were higher comparing...
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Release systems based on self-assembling RADA16-I hydrogels with a signal sequence which improves wound healing processes
PublicationSelf-assembling peptides can be used for the regeneration of severely damaged skin. They can act as scaffolds for skin cells and as a reservoir of active compounds, to accelerate scarless wound healing. To overcome repeated administration of peptides which accelerate healing, we report development of three new peptide biomaterials based on the RADA16-I hydrogel functionalized with a sequence (AAPV) cleaved by human neutrophil elastase...
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Transcriptional activity of epigenetic remodeling genes declines in keratinocytes after in vitro expansion
PublicationPURPOSE: In vitro expansion is an invaluable method to obtain keratinocytes in amounts necessary for effective transplantation therapies. In vitro cell culturing provokes questions concerning potential epigenetic alterations occurring in expanded cells in the context of usefulness for transplantation and safety. The purpose of this study was to investigate as to whether keratinocyte expansion is associated with changes in the activity...
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A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model
PublicationWe have designed a new ddLMS PCR (double digestion Ligation Mediated Suppression PCR) method based on restriction site polymorphism upstream from the specific target sequence for the simultaneous identification and differentiation of bacterial strains. The ddLMS PCR combines a simple PCR used for species or genus identification and the LM PCR strategy for strain differentiation. The bacterial identification is confirmed in the...
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Cross-Linking, Morphology, and Physico-Mechanical Properties of GTR/SBS Blends: Dicumyl Peroxide vs. Sulfur System
PublicationIn this work, ground tire rubber and styrene–butadiene block copolymer (GTR/SBS) blends at the ratio of 50/50 wt%, with the application of four different SBS copolymer grades (linear and radial) and two types of cross-linking agent (a sulfur-based system and dicumyl peroxide), were prepared by melt compounding. The rheological and cross-linking behavior, physico-mechanical parameters (i.e., tensile properties, abrasion resistance,...
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The DraC usher in Dr fimbriae biogenesis of uropathogenic E. coli Dr+ strains
PublicationBiogeneza fimbrii Dr kodowanych przez operon dra uropatogennych szczepów Escherichia coli (infekcje górnych dróg moczowych) odbywa się na bazie zakonserwowanego systemu sekrecji typu chaperone-usher. Funkcjonowanie powyższego systemu sekrecji opiera się na dwóch komponentach białkowych, periplazmatycznym chaperonie DraB i zewnątrzbłonowym kanale DraC. Białko DraB kontroluje proces składania podjednostek fimbrialnych DraE, natomiast...
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The development of an indirect ELISA for the detection of goose parvovirus antibodies using specific VP3 subunits as the coating antigen
PublicationIn Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy’s disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. Prophylactic treatment for DD includes attenuated live or inactivated vaccines. Moreover, the control of DD includes the monitoring of maternal derived antibody (MDA) levels in the offspring and antibody...
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Evaluation of Immobilization of Selected Peat-Isolated Yeast Strains of the Species Candida albicans and Candida subhashii on the Surface of Artificial Support Materials Used for Biotrickling Filtration
PublicationThe paper describes the process of n-butanol abatement by unicellular fungi, able to deplete n-butanol content in gas, by using n-butanol as source of carbon. Isolated and identified fungi species Candida albicans and Candida subhashii were subjected to a viability process via assimilation of carbon from hydrophilic and hydrophobic compounds. The isolates, which exhibited the ability to assimilate carbon, were immobilized on four...
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ZAWARTOŚĆ CUKRÓW W SOKACH DRZEWNYCH Z TERENU PODKARPACIA
PublicationOznaczono zawartość cukrów w sokach drzewnych sześciu gatunków drzew rosnących na terenie Podkarpacia. Materiał doświadczalny stanowiły soki z drzew gatunków: brzoza zwisła in. brodawkowata (Betula pendula Roth.), brzoza omszona (Betula pubescens Ehrh.), grab pospolity (Carpinus betulus L.), klon zwyczajny (Acer platanoides L.), klon jesionolistny (Acer negundo L.) oraz wierzba biała (Salix alba L.). Analizę jakościową cukrów przeprowadzono...
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A Rapid and Simple Method for Screening Microorganisms with a Potential for Catechol Biodegradation
PublicationCatechol is a highly toxic compound that is also a key intermediate in biodegradation pathways of various aromatic compounds. In this paper, a new screening method for isolation of microorganisms with the potential for catechol biodegradation is reported. The method described is based on the ability of catechol to form a color complex with Fe3+ ions. For this purpose, basal medium that is widely used for classification of microorganisms...
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Data regarding a new, vector-enzymatic DNA fragment amplification-expression technology for the construction of artificial, concatemeric DNA, RNA and proteins, as well as biological effects of selected polypeptides obtained using this method
PublicationApplications of bioactive peptides and polypeptides are emerging in areas such as drug development and drug delivery systems. These compounds are bioactive, biocompatible and represent a wide range of chemical properties, enabling further adjustments of obtained biomaterials. However, delivering large quantities of peptide derivatives is still challenging. Several methods have been developed for the production of concatemers –...
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Low frequency of itraconazole resistance found among avian Aspergillus fumigatus isolates
PublicationAspergillus fumigatus is the most frequent etiological factor of invasive aspergillosis in human as well as animals, including birds. Aspergillosis caused by drug-resistant strains poses a significant therapeutic issue, especially if resistance pattern includes insensitivity to triazoles, i.e. drugs recommended for the primary therapy purposes. Majority of resistant isolates hold the TR34/L98H...
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Cold-Active beta-Galactosidases: Sources, Biochemical Properties and Their Biotechnological Potential
Publicationbeta-D-Galactosidases have been studied extensively in terms of their application to a variety of industrial technologies. To date, considerable research efforts have been devoted to characterization of new cold-active beta-D-galactosidases which were isolated directly from selected species of bacteria and yeasts, as well as with the use of metagenomic approaches. This chapter will provide a review of current research towards cold-active...
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Genome-wide DNA methylation profiling of the regenerative MRL/MpJ mouse and two normal strains
PublicationAims We aimed to identify the pivotal differences in the DNA methylation profiles between the regeneration capable MRL/MpJ mouse and reference mouse strains. Materials and Methods Global DNA methylation profiling was performed in ear pinnae, bone marrow, spleen, liver, heart from uninjured adult females of the MRL/MpJ and C57BL/6J and BALB/c. Results and conclusion A number of differentially methylated regions distinguishing between...
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Correlation between the number of Pro-Ala repeats in the EmrA homologue of Acinetobacter baumannii and resistance to netilmicin, tobramycin, imipenem and ceftazidime
PublicationAcinetobacter baumannii coccobacilli are dangerous to patients in intensive care units because of their multidrug resistance to antibiotics, developed mainly in the past decade. This study aimed to examine whether there is a significant correlation between the number of Pro-Ala repeats in the CAP01997 protein, the EmrA homologue of A. baumannii, and resistance to antibiotics. A total of 79 multidrug-resistant A. baumannii strains...
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Genetic Background and Antibiotic Resistance Profiles of K. pneumoniae NDM-1 Strains Isolated from UTI, ABU, and the GI Tract, from One Hospital in Poland, in Relation to Strains Nationally and Worldwide
PublicationIn recent years, there has been an observed increase in infections caused by carbapenem-resistant Klebsiella pneumonia (Kp) strains. The aim of this study was the phenotypic and genotypic analysis of eight K. pneumoniae NDM (Kp NDM) isolates, recovered in Poland during the years 2016 and 2018 from seven patients with urinary tract infections (UTIs), asymptomatic bacteriuria (ABU), or colonization of the gut. PCR melting profile...
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PTD4 Peptide Increases Neural Viability in an In Vitro Model of Acute Ischemic Stroke
PublicationIschemic stroke is a disturbance in cerebral blood flow caused by brain tissue ischemia and hypoxia. We optimized a multifactorial in vitro model of acute ischemic stroke using rat primary neural cultures. This model was exploited to investigate the pro-viable activity of cell-penetrating peptides: arginine-rich Tat(49–57)-NH2 (R49KKRRQRRR57-amide) and its less basic analogue, PTD4 (Y47ARAAARQARA57-amide). Our model included glucose...
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The Immunogenic and Immunoprotective Activities of Recombinant Chimeric T. gondii Proteins Containing AMA1 Antigen Fragments
PublicationToxoplasmosis, one of the most common parasitoses worldwide, is potentially dangerous for individuals with a weakened immune system, but specific immunoprophylaxis intended for humans is still lacking. Thus, efforts have been made to create an efficient universal vaccine for both animals and humans to overcome the shortcomings of currently used treatment methods and protect all hosts against toxoplasmosis. The current work represents...
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The Effect of Posaconazole, Itraconazole and Voriconazole in the Culture Medium on Aspergillus fumigatus Triazole Resistance
PublicationTriazoles are the only compounds used as antibiotics in both medicine and agriculture. The presence of triazoles in the environment can contribute to the acquisition of azole resistance among isolates of Aspergillus fumigatus. The objective of this study was to investigate the effect of A. fumigatus exposure to triazoles on susceptibility to these compounds. Seventeen triazole-resistant and 21 triazole-sensitive A. fumigatus isolates...
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Amyloid fibril formation in the presence of water structure-affecting solutes
PublicationThe impact of the differently hydrated non-electrolytes (protein structure destabilizers) on the fibrillation of hen egg white lysozyme (HEWL) was investigated. Two isomeric urea derivatives i.e. butylurea (BU) and N,N,N′,N′-tetramethylurea (TMU) were chosen as a tested compounds. The obtained results show that butylurea exerts greater impact on HEWL and its fibrillation than tetramethylurea. Both substances decrease the time of...
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Transcriptomic Effects of Estrogen Starvation and Induction in the MCF7 Cells. The Meta-analysis of Microarray Results
PublicationEstrogen is one of the most important signaling molecules which targets a number of genes. Estrogen levels regulate cell proliferation and a plethora of metabolic processes, which may interfere with a range of medical conditions and drug metabolism. The MCF7 breast cancer cell line, expressing the estrogen receptor α, is a well-studied model of cellular answer to estrogen. The aim of this study was to characterize transcriptomic...
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A simple modification to improve the accuracy of methylation-sensitive restriction enzyme quantitative polymerase chain reaction
PublicationDNA digestion with endonucleases sensitive to CpG methylation such as HpaII followed by polymerase chain reaction (PCR) quantitation is commonly used in molecular studies as a simple and inexpensive solution for assessment of region-specific DNA methylation. We observed that the results of such analyses were highly overestimated if mock-digested samples were applied as the reference.We determined DNA methylation levels in several...
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Functionalized Peptide Fibrils as a Scaffold for Active Substances in Wound Healing
PublicationTechnological developments in the field of biologically active peptide applications in medicine have increased the need for new methods for peptide delivery. The disadvantage of peptides as drugs is their low biological stability. Recently, great attention has been paid to self-assembling peptides that can form fibrils. Such a formulation makes bioactive peptides more resistant to enzymatic degradation and druggable. Peptide fibrils...
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Escherichia coli Strains with Virulent Factors Typical for Uropathogens were Isolated from Sinuses from Patients with Chronic Rhinosinusitis—Case Report
PublicationEscherichia coli were isolated from three patients with chronic rhinosinusitis (CRS) by intraoperative sinus tissue biopsy. Taking into account the unusual replicative niche and previous treatment failures, it was decided to focus on the virulence and drug resistance of these bacteria. The strains turned out to be multi-sensitive, but the rich virulence factors profile of bacteria typical for phylogenetic group B2 deserved attention....
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The extended version of restriction analysis approach for the examination of the ability of low-molecular-weight compounds to modify DNA in a cell-free system
PublicationOne of the primary requirements in toxicology is the assessment of ability of chemicals to induce DNA covalent modification. There are several well-established methods used for this purpose such as 32P-Postlabeling or HPLC-MS. However, all of these approaches have difficult to overcome limitations, which prevents their use in genotoxin screening. Here, we describe the simple protocol exploiting specificity of restriction enzymes...
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In Situ Random Microseeding and Streak Seeding Used for Growth of Crystals of Cold-Adapted beta-D-Galactosidases: Crystal Structure of betaDG from Arthrobacter sp. 32cB
PublicationThere is an increasing demand for cold-adapted enzymes in a wide range of industrial branches. Nevertheless, structural information about them is still scarce. The knowledge of crystal structures is important to understand their mode of action and to design genetically engineered enzymes with enhanced activity. The most difficult task and the limiting step in structural studies of cold-adapted enzymes is their crystallization,...
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A Colorimetric Microplate Assay for DNA-Binding Activity of His-Tagged MutS Protein
PublicationA 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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Recurrent bowel-blood translocations of Escherichia coli with the unique virulence characteristics over three-year period in the patient with acute myeloid leukaemia – case report
PublicationIn patients with haematological malignancies, the bowel remains the main source of Escherichia coli bloodstream infections. We present the clinical example of recurrent bowel-blood translocations of E. coli with the unique virulence characteristics in a 55-year-old male with the diagnosis of acute myeloid leukaemia. The virulent factors profile of examined strains confirmed that the co-existence of genes papC, sfa, usp and cnf1,...
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Borrelia burgdorferi BmpA-BBK32 and BmpA-BBA64: New Recombinant Chimeric Proteins with Potential Diagnostic Value
PublicationCurrently, the diagnosis of Lyme disease is based mostly on two-tiered serologic testing. In the new generation of immunoenzymatic assays, antigens comprise whole-cell lysates of members of the Borrelia burgdorferi sensu lato (s.l.) species complex, with the addition of selected recombinant proteins. Due to the high diversity of members of the B. burgdorferi s.l. genospecies and the low degree of conservation among the amino acid...
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Removal of cyclohexane and ethanol from air in biotrickling filters inoculated with Candida albicans and Candida subhashii
PublicationThis paper presents investigations on the removal of cyclohexane and ethanol from air in polyurethane- -packed biotrickling filters, inoculated with Candida albicans and Candida subhashii fungal species. Results on process performance together with flow cytometry analyses of the biofilm formed over packing elements are presented and discussed. The results indicate that the presence of ethanol enhances the removal efficiency of cyclohexane...
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Identification and characterization of single-stranded DNA-binding protein from the facultative psychrophilic bacteria Pseudoalteromonas haloplanktis
PublicationSingle-stranded DNA-binding protein (SSB) plays an important role in DNA metabolism such as DNAreplication, repair, and recombination, and is essential for cell survival. This study reports on the ssb-likegene cloning, gene expression and characterization of a single-stranded DNA-binding protein of Pseudoal-teromonas haloplanktis (PhaSSB) and is the first report of such a protein from psychrophilic microorganism.PhaSSB possesses...
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PCR and real-time PCR approaches to the identification of Arthroderma otae species Microsporum canis and Microsporum audouinii/Microsporum ferrugineum
PublicationObjectives The identification of species in the Arthroderma otae complex is essential to determine the origin of infection and to eliminate the risk of transmission. Microsporum canis is a zoophilic species, whereas Microsporum audouinii and Microsporum ferrugineum are anthropophilic species. In this paper, we propose alternative methods that permit species-specific identification of both anthropophilic and zoophilic members of...
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Urinary Tract Infections Caused by K. pneumoniae in Kidney Transplant Recipients – Epidemiology, Virulence and Antibiotic Resistance
PublicationUrinary tract infections are the most common complication in kidney transplant recipients, possibly resulting in the deterioration of a long-term kidney allograft function and an increased risk of recipient’s death. K. pneumoniae has emerged as one of the most prevalent etiologic agents in the context of recurrent urinary tract infections, especially with multidrug resistant strains. This paper discusses the epidemiology and risk...
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Application of safirinium N-hydroxysuccinimide esters to derivatization of peptides for high-resolution mass spectrometry, tandem mass spectrometry, and fluorescent labeling of bacterial cells
PublicationMass spectrometry methods are commonly used in the identification of peptides and biomarkers. Due to a relatively low abundance of proteins in biological samples, there is a need for the development of novel derivatization methods that would improve MS detection limits. Hence, novel fluorescent N-hydroxysuccinimide esters of dihydro-[1,2,4]triazolo[4,3-a]pyridin-2-ium carboxylates (Safirinium P dyes) have been synthesized. The...
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Characterization of d-xylose reductase, XyrB, from Aspergillus niger
PublicationD-xylose reductase is a member of the aldo-keto reductase family, and is involved in D-xylose and L-arabinose conversion through the Pentose Catabolic Pathway (PCP) in fungi. In this study, we biochemically characterized a newly identified second D-xylose reductase (XyrB) from Aspergillus niger. This NADPH-dependent reductase is able to efficiently convert D-xylose and L-arabinose, and it has the highest affinity for these sugars...
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Polymer biodegradable coatings as active substance release systems for urological applications
PublicationCatheter-associated urinary tract infections are the result of catheterization of the bladder. The risk of infection is directly proportional to the length of the bladder catheterization. Bacteria cells have the ability to adhere and create the biofilm on the surface of catheter materials. Bacteria’s biofilm is an extremely beneficial environment of existence for microorganisms. Microorganisms that are an integral part of the biofilm...
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Evaluation of the content of inorganic anions in tree saps
PublicationTree saps were once commonly used in the countries of Northern, Central and Eastern Europe. Although once almost forgotten, their popularity has been growing recently as part of an interest in organic food and traditional medicine. Tree saps, tapped mainly from birch and maple trees, are drunk both fresh and fermented or are used as raw material for the production of food products, e.g. syrups. The aim of this study was to determine...
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IgG Avidity Test as a Tool for Discrimination between Recent and Distant Toxoplasma gondii Infection—Current Status of Studies
PublicationToxoplasma gondii, an obligate intracellular protozoan parasite, is the causative agent of one of the most prevalent zoonoses worldwide. T. gondii infection is extremely important from a medical point of view, especially for pregnant women, newborns with congenital infections, and immunocompromised individuals. Thus, an accurate and proper diagnosis of this infection is essential. Among the available diagnostic tests, serology...