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Search results for: TAQ DNA POLYMERASE, GC-RICH TEMPLATES, PCR
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Diagnostic PCR assay for Microsporum and Trichophyton infections
PublicationWe 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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Optimized 5-hour multiplex PCR test for the detection of tinea unguium: performance in a routine PCR laboratory
PublicationWe recently reported the development of a 5-hour multiplex PCR test for the detection of tinea unguium and the optimization of this test by the inclusion of an inhibition control. Here we report the performance of this procedure as used in a routine clinical laboratory as compared to conventional microscopy and culture-based techniques performed in a mycology reference laboratory. We found in processing 109 samples that 22 (20.2%)...
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Five-hour diagnosis of dermatophyte nail infections with specific detection of Trichophyton rubrum
PublicationW publikacji przedstawiono szybką dwuetapową metodę oczyszczania DNA oraz opartą na technice multiplex PCR metodę wykrywania grzybów z rodzaju dermatofitów oraz gatunku Trichophyton rubrum z hodowli grzybów oraz bezpośrednio ze zmienionych chorobowo paznokci. Testy stu osiemnastu próbek z użyciem zaprojektowanej reakcji multiplex PCR i dwóch kolejnych (jedna wykrywająca grzyby z rodzaju dermatofitów, druga wykrywająca gatunek T....
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ADSRRS-fingerprinting and PCR MP techniques for studies of intraspecies genetic relatedness in Staphylococcus aureus
PublicationW pracy pokazano użyteczność dwóch nowych metod do typowania molekularnego Staphylococcus aureus, ADSRRS-fingerprinting (Amplification of DNA fragments Surrounding Rare Restriction Sites) i PCR MP (PCR melting profile). Przebadano 37 szczepów Staphylococcus aureus izolowanych od pacjentów z czyracznością. Jako metodę referencyjną zastosowano REA-PFGE. Okazało się, że izolaty Staphylococcus aureus z nosa od niektórych pacjentów...
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Optimised five-hour multiplex PCR test for the detection of Tinea ungium: performance in a routine PCR laboratory
PublicationWe recently published the development of a 5-hour multiplex PCR test for the detection of dermatophyte nail infection. We have optimized this test by inclusion of an inhibition control and evaluated the test in a routine laboratory when compared to the conventional microscopy and culture. A total of 109 clinical samples received at the mycology reference lab at Statens Serum Institute were included. The samples were divided equally...
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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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Genotyping Techniques for Determining the Diversity of Microorganisms
PublicationTyping of microbial pathogens, 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. The part...
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Therapeutic intervention by the simultaneous inhibition of DNA repair and type I or type II DNA topoisomerases: one strategy, many outcomes
PublicationMany anticancer drugs reduce the integrity of DNA, forming strand breaks. This can cause mutations and cancer or cell death if the lesions are not repaired. Interestingly, DNA repair-deficient cancer cells (e.g., those with BRCA1/2 mutations) have been shown to exhibit increased sensitivity to chemotherapy. Based on this observation, a new therapeutic approach termed 'synthetic lethality' has been developed, in which radiation...
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5-Bromo-2′-deoxycytidine—a potential DNA photosensitizer
PublicationA double-stranded oligonucleotide, 80 base pairs in length, was multiply labeled with 5-bromo-2′-deoxycytidine (BrdC) using polymerase chain reaction (PCR). The modified oligonucleotide was irradiated with 300 nm photons and its damage was assayed by employing DHPLC, LC-MS and denaturing polyacrylamide gel electrophoresis (PAGE). Two types of damage were demonstrated, namely, single strand breaks (SSBs) and intrastrand cross-links...
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Evaluation of possibilities of genotyping of Candida glabrata clinical isolates with RAPD-PCR method and microsatellite analysis
PublicationThe aim of the study was to compare the discriminatory power of RAPD-PCR method using RSD10 primer and microsatellite (GACA)4 analysis for genotyping clinical isolates of C. glabrata. Isolates were received from patients of two Polish hospitals: Children's Memorial Health Institute in Warsaw (n=17) and Medical University of Gdansk (n=37). Species identification was confirmed by two phenotypic methods: growth on chromagar plates...
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Real-time PCR approach in dermatophyte detection and Trichophyton rubrum identification
PublicationDermatophytes are keratinophilic molds that infect human hair, nails and skin. Diagnosis of dermatophytosis is based on morphological, serological and biochemical features. However, identification is difficult and laborious due to similarities between microorganisms. Thus, there is considerable interest to develop mycological diagnostic procedures based on molecular biology methods. In this study, fast, two-step DNA extraction...
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Evaluation of a PCR Melting Profile method for intra-species differentiation of Trichophyton rubrum and Trichophyton interdigitale
PublicationW celu identyfikacji źródła infekcji spowodowanych przez dermatofity, jak i drogi ich transmisji, istnieje potrzeba szukania nowych metod, które pozwolą na głębokie zróżnicowanie genetyczne szczepów w obrębie rodzaju. W pracy po raz pierwszy pokazano zastosowanie techniki PCR MP opartej o adaptory oligonukleotydowe i różnice w temperaturze topnienia fragmentów restrykcyjnych DNA w reakcji PCR do wewnątrzgatunkowego typowania dermatofitów....
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System do diagnostyki dermatofitowych zakażeń powierzchniowych oparty na technice Real-Time PCR
PublicationDermatofity należą do blisko spokrewnionej grupy grzybów, które wykazują wysokie powinowactwo do skeratynizowanych tkanek. Cecha ta czyni je odpowiedzialnymi za powierzchniowe grzybice skóry, paznokci oraz włosów. Szacuje się, iż nawet do 20% ludzi na całym świecie dotkniętych jest infekcjami powodowanymi przez dermatofity. Ich leczenie wymaga długotrwałego zastosowania leków przeciwgrzybiczych. W celu dobrania odpowiedniego leczenia...
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Construction and purification of his6-Thermus thermophilus MutS protein.
PublicationGen mutS z termofilnej bakterii Thermus thermophilus zamplifikowano przy użyciu PCR, sklonowano i przeprowadzono jego ekspresję w E. coli. Rekombinantowe białko MutS zawierające oligohistydynową domenę na N-końcu oczyszczono w jednoetapowej procedurze przy wykorzystaniu chromatografii powinowactwaNi(2+). Zdolność do rozpoznania niekomplementarności białka his(6)-MutS potwierdzono w eksperymentach ochrony DNA przed trawieniem...
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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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Clinical studies of detecting COVID-19 from exhaled breath with electronic nose
PublicationThe COVID‑19 pandemic has attracted numerous research studies because of its impact on society and the economy. The pandemic has led to progress in the development of diagnostic methods, utilizing the polymerase chain reaction (PCR) as the gold standard for coronavirus SARS‑CoV‑2 detection. Numerous tests can be used at home within 15 min or so but of with lower accuracy than PCR. There is still a need for point‑of‑care tests available...
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Discrimination of hospital isolates of Acinetobacter baumannii using repeated sequences and whole genome alignment differential analysis
PublicationAn optimized method for bacterial strain differentiation, based on combination of Repeated Sequences and Whole Genome Alignment Differential Analysis (RS&WGADA), is presented in this report. In this analysis, 51 Acinetobacter baumannii multidrug-resistance strains from one hospital environment and patients from 14 hospital wards were classified on the basis of polymorphisms of repeated sequences located in CRISPR region, variation...
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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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UV-induced electron transfer between triethylamine and 5-bromo-2′-deoxyuridine. A puzzle concerning the photochemical debromination of labeled DNA
Publication5-Bromo-2'-deoxyuridine (BrdU) photosensitizes DNA to strand break formation. However, this type of photodamage is completely quenched by the presence of triethylamine (TEA) which originates from RP-HPLC purification commonly employed by oligonucleotide providers. While the presence of TEA in oligonucleotide samples does not interfere with PCR or other molecular biology applications, the mechanism of photochemical reaction proceeding...
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Experimental guinea pig model of dermatophytosis: a simple and useful tool for the evaluation of new diagnostics and antifungals
PublicationCelem badań było opracowanie prostego modelu zwierzęcego (świnki morskie) odpowiedniego do sprawdzania systemów diagnostycznych oraz terapii infekcji dermatofitów. Następujące zmienne były analizowane: przygotowanie skóry przez golenie versus użycie taśmy, testowy szczep Microsporum canis czy Trichophyton mentagrophytes jako czynnik etiologiczny, sposób inokulacji. Przebieg infekcji był oceniany poprzez ocenę zaczerwienienia i...
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Genotyping and characterization of virulence factors in Escherichia coli strains isolated from patients from Polish hospital
PublicationZakażenia układu moczowego (UTIs - ang. urinary track infections) są jedną z najczęstszy infekcji, za które w 85% przypadków odpowiedzialne są bakterie Escherichia coli. Infekcje układu moczowego mogą prowadzić do groźnych powikłań m. in. odmiedniczkowego zapalenia nerek. Śmiertelność wśród tej grupy pacjentów wynosi 1-3%. Zakażenia układu moczowego występują również u ok. 7% kobiet ciężarnych. W pracy przebadano 82 szczepy E....
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Analysis and characterization of coordination compounds by resonance Raman spectroscopy
PublicationResonance Raman spectroscopy has become a powerful tool to study excited-state geometries, excited-state charge distributions and photoinduced reaction dynamics in coordination compounds. Due to their rich photophysical properties coordination compounds are utilized for a variety of applications ranging from DNA sensing to photocatalysis. This review features recent applications of various resonance Raman scattering techniques...
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New Approaches for Escherichia coli Genotyping
PublicationEasy-to-perform, fast, and inexpensive methods of differentiation of Escherichia coli strains beyond the species level are highly required. Herein two new, original tools for genotyping of E. coli isolates are proposed. The first of the developed method, a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) test uses a highly variable fliC gene, encoding the H antigen as a molecular target. The designing...
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Fungal Typin Methods
PublicationThe broad application of the molecular techniques in mycoses diagnosis is related to increase of infections caused by fungi in many countries. The oldest typing methods relaying on fenotypic observation, physiological and biochemical examination have had very limited importance from decades. Novadays, the molecular biology methods took their place. Most of the genotyping methods have been devoleped to be applied for typing of bacteria...
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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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PCR detection of Scopulariopsis brevicaulis
PublicationScopulariopsis brevicaulis is known as a most common etiological factor of the mould toenail infections. There are also reports indicating that S. brevicaulis could cause organ and disseminated infections. Nowadays microscopic observations from the direct sample and culture are crucial for the appropriate recognition of the infection. In this paper is presented a PCR-based method for S. brevicaulis detection. The specificity of...
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Isolation and identification of Mycobacterium avium subspecies silvaticum from a horse
PublicationPo raz pierwszy opracowano metodę identyfikacji Mycobacterium avium subsp. silvaticum na podstawie analizy sekwencji DNA alfa antygenu. Zastosowanie enzymu HhaI do trawienia produktu PCR z uzyskanego z udziałem starterów specyficznych wobec genu kodującego alfa antygen pozwoliło na identyfikację Mycobacterium avium subsp. silvaticum. Opracowana nowa metoda pozwala na prowadzenie badań epidemiologicznych bezpośrednio z próbek pochodzących...
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The use of a one-step PCR method for the identification of Microsporum canis and Trichophyton mentagrophytes infection of pets
PublicationIntroduction: Dermatophytes are a closely related group of keratinophilic fungi. They encompass important etiological agents of superficial fungal infections. These fungi are able to invade keratinized tissues of humans and animals, causing dermatophytosis (ringworm) of hair, nails or skin. The aim: Traditional diagnostics of ringworm is based on morphological identification of cultured fungi and is time-consuming. Materials and methods:...
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B-GALACTOSIDASE ARTHROBACTER SP. 32cB - OBTAINING THE GENE SEQUENCE, CONSTRUCTION OF THE EXPRESSION SYSTEM, BIOSYNTHESIS AND BIOCHEMICAL CHARACTERIZATION OF THE ENZYME
PublicationINTRODUCTION: β-Galactosidase is an enzyme which catalyzes the hydrolysis of O glycosidic bond in β-galactosides. Another activity of β galactosidase is a transglycosylation activity. The main industrial use of this protein is the hydrolysis of lactose in milk in a cooling conditions. Synthesis of galactooligosaccharides, which are mostly used as a prebiotics added to some foods or available as dietary supplements, is only one...
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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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DNA restriction analysis as a supportive tool in mechanistic studies carried out by 32P-postlabelling
PublicationNumerous antitumor and carcinogenic compounds are able to modify DNA by forming covalent bonds with its constituents, while some anticarcinogenic compounds are known to prevent such a modification. All these processes are of vital biological import_ance, though deeper inside into factors influencing formation of DNA adducts is difficult due to the low level of their occurrence. 32P-Postlabelling approach ensures very sensitive...
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Motility activity, slime production, biofilm formation and genetic typing by ERIC-PCR for Pseudomonas aeruginosa strains isolated from bovine and other sources (human and environment)
PublicationThe molecular-typing strategy, ERIC-PCR was used in an attempt to determine the genomic relationship of 28 P. aeruginosa strains isolated from faeces of healthy bovine, bovine mastitis and from faeces of hospital patients as well as from environment. ERIC-PCR fingerprinting revealed large molecular differentiation within this group of isolates. Twenty two out of 28 strains tested generated unique patterns of DNA bands and only...
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PARP inhibition potentiates the cytotoxic activity of C-1305, a selective inhibitor of topoisomerase II, in human BRCA1-positive breast cancer cells
PublicationTwo cellular proteins encoded by the breast and ovarian cancer type 1 susceptibility (BRCA1 and BRCA2) tumor suppressor genes are essential for DNA integrity and the maintenance of genomic stability.Approximately 5-10% of breast and ovarian cancers result from inherited alterations or mutations in these genes.Remarkably, BRCA1/BRCA2-deficient cells are hypersensitive to selective inhibition of poly(ADPribose) polymerase 1 (PARP-1),...
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Typing of Scopulariopsis and Microascus fungi by Random Amplified Polymorphic DNA (RAPD)
PublicationOBJECTIVES Scopulariopsis species and their teleomorphs of the genus Microascus are commonly isolated from soil, decaying plant material and indoor environments. Moreover, certain Scopulariopsis and Microascus species are recognised as opportunistic human pathogens. Although most species can be identified by detailed morphological study, phenotypic characters appear to overlap in several cases and morphology seems to be insufficient...
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Two highly thermostable paralogous single-stranded DNA-binding proteins from Thermoanaerobacter tengcongensis
PublicationThe thermophilic bacterium Thermoanaerobacter tengcongensis has two single-stranded DNA-binding (SSB) proteins, designated TteSSB2 and TteSSB3. In a SSB complementation assay in Escherichia coli, only TteSSB3 took over the in vivo function of EcoSSB. We have cloned the ssb genes obtained by PCR and have developed E. coli overexpression systems. The TteSSB2 and TteSSB3 consist of 153 and 150 amino acids with a calculated molecular...
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Conserved motifs of MutL proteins
PublicationtThe MutL protein is best known for its function in DNA mismatch repair (MMR). However, there isevidence to suggest that MutL is not only the linker connecting the functions of MutS and MutH in MMR,but that it also participates in other repair systems, such as Very Short Patch (VSP), Base Excision (BER)and Nucleotide Excision Repair (NER). This study set out to identify the most highly conserved aminoacid sequence motifs in MutL...
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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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Resistance of Escherichia coli and Enterococcus spp. to selected antimicrobial agents present in municipal
PublicationIn this study, the susceptibility to erythromycin (E) and to trimethoprim/sulfamethoxazole (SXT) among isolates of Enterococcus spp. and Escherichia coli was tested, respectively. Both fecal indicators were detected and isolated from raw(RW) and treated wastewater (TW) as well as from samples of activated sludge (AS) collected in a local wastewater treatment plant (WWTP). Biodiversity of bacterial community in AS was also monitored...
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Novel single-stranded DNA-binding protein from psyvhrophilic bacterium Psychrobacter arcticus
PublicationTo study the biochemical properties of SSB from Psychrobacter arcticus (ParSSB), we have cloned the ssb genes obtained by PCR and have developed Escherichia coli overexpression systems. The gene consists of an open reading frame of 642 nucleotides encoding SSB protein of 213 amino acids with a calculated molecular mass of 22.8 kDa. The amino-acid sequence of ParSSB exhibits 49% identity and 57% similarity to Escherichia coli SSB....
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Tetracycline and ampicillin resistance of Escherichia coli strains of surface water origin: the potential for horizontal transfer of resistance genes = Oporność na tetracyklinę i ampicylinę szczepów Escherichia Coli wyizolowanych z wody powierzchniowej : możliwość horyzontalnego przenoszenia genów oporności
PublicationThe aim of this preliminary study was to assess the occurrence and molecular diversity of tetracycline and ampicillin resistance genes carried by Escherichia coli present in surface water. Bacterial strains were isolated from two watercourses (the Reda River and the Oliwski Stream) that influence the quality of coastal waters in the Gdańsk Bay (Northern Poland) by direct discharge. The bacterial drug susceptibility, tested against...
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DNA adduct formation by C-1748, a potent antitumor 4-methyl-1-nitroacridine of lowered toxicity.
Publication4-podstawione 1-nitroakrydyny reprezentują nową grupę pochodnych zsyntetyzowanych na Politechnice Gdańskiej. W porównaniu do macierzystych 1-nitroakrydyn związki te wykazują obniżoną toksyczność i podwyższoną aktywność przeciwnowotworową. Wiodąca pochodna 4-metylo-1-nitroakrydyna o symbolu C-1748 jest obecnie w przygotowaniu do I fazy badań klinicznych w leczeniu raka okrężnicy. Wprowadzenie elektrodonorowej grupy metylowej w...
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Impact of pollution derived from ship wrecks on the marine environment on the basis of s/s ''Stuttgart'' (Polish coast, Europe)
PublicationW 1943 roku niemiecki statek szpitalny, s/s Stuttgart (Lazaretschiff ''C'') został zatopiony w pobliżu portu w Gdyni. Doprowadziło to do zanieczyszczenia dna morskiego paliwem. Podczas badań przeprowadzonych w 2009 r. pobranych zostało 11 próbek osadów powierzchniowych i wody morskiej oraz 4 rdzenie. Próbki poddano analizie na zawartość zanieczyszczeń priorytetowych należących do grupy wielopierścieniowych węglowodorów aromatycznych...
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Genome-Wide DNA Methylation and Gene Expression in Patients with Indolent Systemic Mastocytosis
PublicationMastocytosis is a clinically heterogenous, usually acquired disease of the mast cells with a survival time that depends on the time of onset. It ranges from skin-limited to systemic disease, including indolent and more aggressive variants. The presence of the oncogenic KIT p. D816V gene somatic mutation is a crucial element in the pathogenesis. However, further epigenetic regulation may also affect the expression of genes that...
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Rozdział 5. Nowoczesne metody diagnostyki zakażeń grzybiczych wywoływanych przez dermatofity = Novel diagnostic methods of funfal infections caused by dermatophytes
PublicationDermatofity są głównym czynnikiem etiologicznym grzybic powierzchownych. Do tej klasy grzybów należą przedstawiciele trzech rodzajów: Epidermophyton, Microsporum i Trichophyton. Klasyczne metody diagnostyczne polegają na bezpośredniej ocenie mikroskopowej oraz badaniu hodowlanym. Identyfikacja gatunkowa jest możliwa na podstawie cech makroskopowych kolonii: ich struktury, koloru, kształtu, oraz obserwacji mikroskopowych (kształt...
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Host and pathogen factors in Klebsiella pneumoniae upper urinary tract infections in renal transplant patients
PublicationPurpose . To analyse the role of virulence factors (VFs) and host in Klebsiella pneumoniae upper urinary tract infections (UTIs) in renal transplant (RTx) recipients. Methodology. Clinical and demographic data were registered prospectively. Phylogenetic background of K. pneumoniae isolates was analysed by PCR melting profiles (MP) and the following VFs genes: fimH-1, uge, kpn, ycfM, mrkD, rmpA, magA, hlyA, cnf-1, irp-1, irp-2,...
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Identification and properties of the psychrophilic bacterium Desulfolatea psychrophila single-stranded DNA binding proteins
PublicationTo study the biochemical properties of SSB from Desulfolatea psychrophila (DpsSSB), we have cloned the ssb genes obtained by PCR and have developed Escherichia coli overexpression systems. The gene consists of an open reading frame of 426 nucleotides encoding SSB protein of 142 amino acids with a calculated molecular mass of 15.6 kDa and it is the smallest known bacterial SSB protein indentified to date. The amino-acid sequence...
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Novel staphylococci nucH taxonomical marker used in identification of human-associated Staphylococcus succinus subsp. casei
PublicationThe aim of our study was to assess the sequencing of unique nucH gene fragment based on performed bioinformatics analysis as a novel diagnostic method for the identification of difficult to identify staphylococcal human pathogenic strains. Initially, PCR-RFLP-rrn analysis specific to the spacers between 16SrDNA and 23SrDNA followed by HhaI restriction analysis was performed. Further, sequencing of nucH and 16S rDNA genes fragments...
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Nowoczesne metody diagnostyki zakażeń dermatofitowych
PublicationGrzyby należące do rodzajów Microsporum, Trichophyton i Epidermophyton są główną przyczyną powierzchownych zakażeń grzybiczych. Klasyczna identyfikacja gatunkowa czynnika etiologicznego tych grzybic oparta na obserwacjach mikro- i makroskopowych możliwa jest dopiero po uzyskaniu czystej hodowli dermatofitów. W związku z powolnym tempem wzrostu jest ona możliwa po dwóch, trzech tygodniach po pobrania próbki. Molekularne metody diagnostyczne...
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A highly thermostable, homodimeric single-stranded DNA-binding protein from Deinococcus radiopugnans
PublicationWe report the identification and characterization of the single-stranded DNA-binding protein (SSB) from the mesophile and highly radiation-resistant Deinococcus radiopugnans (DrpSSB). PCR-derived DNA fragment containing the complete structural gene for DrpSSB protein was cloned and expressed in Escherichia coli. The gene consisting of an open reading frame of 900 nucleotides encodes a protein of 300 amino acids with a calculated...
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C-1311 (Symadex), a potential anti-cancer drug, intercalates into DNA between A and G moieties. NMR-derived and MD-refined stereostructure of the d(GAGGCCTC) 2 :C-1311 complex
PublicationImidazoacridinone C-1311 (Symadex®) is an antitumor agent which has been recommended for Phase II clinical trials a few years ago. Previously, it was shown experimentally that during the initial stage of its action C-1311 forms stable intercalation complexes with DNA duplexes. Herein, a NMR-derived stereostructure of d(GAGGCCTC)2:C-1311 complex was reported. The ligand was found locating itself between A and G moieties, forming...