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Katalog Publikacji
Rok 2015
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Taq DNA polymerase fused with DNA binding protein with increased resistance to inhibitors from clinical samples
PublikacjaNowadays PCR method is commonly used in molecular diagnostic. However, in many cases PCR is limited, by the presence of inhibitory substances in biological, soil or food samples Efficiency and fidelity of amplification is strongly connected with DNA polymerase and reaction conditions. To meet the requirements of modern diagnostic methods it is essential to seeking for new DNA polymerases with better properties useful in these field....
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Specific detection of Alternaria alternata by PCR and real-time PCR
PublikacjaFungi of Alternaria genus are cosmopolitan organisms, which spores can be found in the air, soil, water, clothing and food. They commonly occur as saprotrophs on the plant remains, contributing to the decomposition of organic matter. Additionally, they are components of the normal human and animal skin flora. Alternaria spp. are also known human allergens, causing hay fever and allergic reactions that can lead to the development...
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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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Real-time PCR approach in dermatophyte detection and Trichophyton rubrum identification
PublikacjaDermatophytes 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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Prime-Boost Vaccination with Toxoplasma Lysate Antigen, but Not with a Mixture of Recombinant Protein Antigens, Leads to Reduction of Brain Cyst Formation in BALB/c Mice
PublikacjaIntroduction Infection with the ubiquitous parasite Toxoplasma gondii is a threat for immunocompro- mised patients and pregnant women and effective immune-prophylaxis is still lacking. Methods Here we tested a mixture of recombinant T . gondii antigens expressed in different develop- mental stages, i.e., SAG1, MAG1 and GRA7 (SMG), and a lysate derived from T . gondii tachyzoites (TLA) for prophylactic vaccination against cyst formation....
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PCR detection of Scopulariopsis brevicaulis
PublikacjaScopulariopsis 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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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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Novel application of terminal restriction fragments length polymorphism method (t-RFLP) for identification of six clinically important Candida sp.
PublikacjaTerminal restriction fragments length polymorphism method (t-RFLP) is a modified classical RFLP method. The main difference is based on carrying out PCR using a fluorescently labeled primer or primers. The result of such modification is that after digestion of amplicons only terminal fragments of PCR product (containing fluorescent dye) are visualized. T-RFLP method has found application in environmental microbiology and epidemiology....
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New recombinant chimeric antigens, P35-MAG1, MIC1-ROP1, and MAG1-ROP1, for the serodiagnosis of human toxoplasmosis
PublikacjaThe aim of the study was to evaluate the usefulness of 3 chimeric Toxoplasma gondii antigens, P35-MAG1,MIC1-ROP1 andMAG1-ROP1, in the serodiagnosis of an acute toxoplasmosis in humans. Proteins were produced as fusion proteins containing His tags ends and then further purified by metal affinity chromatography. Their application for the diagnosis of recently acquired T. gondii infection was tested in IgG and IgM enzyme-linked immunosorbent...
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Mechanisms of azole resistance among clinical isolates of Candida glabrata in Poland
PublikacjaCandida glabrata is currently ranked as the second most frequently isolated aetiological agent of human fungal infections, next only to Candida albicans. In comparison with C. albicans, C. glabrata shows lower susceptibility to azoles, the most common agents used in treatment of fungal infections. Interestingly, the mechanisms of resistance to azole agents in C. albicans have been much better investigated than those in C. glabrata....
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Increased concentration of Taq DNA polymerase as a solution for GC-rich templates from clinical and environmental samples
PublikacjaDNA polymerase is an enzyme which plays crucial role in replication and DNA repair. It found application in PCR (polymerase chain reaction) where catalyses process of in vitro DNA synthesis. To meet the demands posed by mod- ern diagnostic, molecular biology or genetic engineering it is necessary to improve DNA polymerases to obtain new or better features useful in these fields. So far implemented modifications in majority are...
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Identification of antigen Ag43 in uropathogenic Escherichia coli Dr+ strains and defining its role in the pathogenesis of urinary tract infections
PublikacjaUrinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) are among the most common bacterial infectious diseases in the developed world. The urovirulence of UPEC is mainly associated with the surface-exposed fimbrial adhesins and adhesins of the autotransporter (AT) family. The best studied of theses proteins is antigen 43 (Ag43) mediating cell aggregation, adhesion and biofilm development as the causes of...
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First report of seroprevalence of Toxoplasma gondii infection in sheep in Pomerania, northern Poland
PublikacjaIntroduction and objective. Toxoplasmosis is parasitic disease which has economic relevance for both veterinary and human medicine. In sheep, toxoplasmosis is a major cause of abortion and can thus cause reproductive problems. The current study aimed to determine the occurrence of anti-Toxoplasma gondii IgG antibodies in sheep from 13 districts of northern Poland and thereby obtain actual data about T. gondii seroprevalence in...
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Expression of goose parvovirus whole VP3 protein and its epitopes in Escherichia coli cells
PublikacjaThe 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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Examination of cyp51A and cyp51B expression level of the first Polish azole resistant clinical Aspergillus fumigatus isolate
PublikacjaAspergillus fumigatus is one of the most prevalent airborne fungal pathogens causing infections worldwide. Most A. fumigatus strains are susceptible to azoles, which are administered as the first line therapeutics. However, during last decade the acquired resistance to triazoles by these species has been described. There is a number of publications concerning the examination of clinical A. fumigatus strains from different countries,...
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Essential Oils, Silver Nanoparticles and Propolis as Alternative Agents Against Fluconazole Resistant Candida albicans, Candida glabrata and Candida krusei Clinical Isolates
PublikacjaDevelopment of effective and safe therapeutic treatment of fungal infections remains one of the major challenge for modern medicine. The aim of presented investigation was to analyze the in vitro antifungal activity of selected essential oils, ethanolic extracts of propolis and silver nanoparticles dropped on TiO2 against azole-resistant C. albicans (n = 20), C. glabrata (n = 14) and C. krusei (n = 10) clinical isolates. Among...
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Epigenetyczne i transkryptomiczne podstawy potencjału regeneracyjnego ssaków. Badania na modelach skóry płodowej i dorosłej myszy MRL/MpJ
PublikacjaRegeneracja jest to zdolność do odbudowy uszkodzonych lub utraconych tkanek, kończyn oraz narządów. Efektywne procesy regeneracji obserwowane są u prymitywnych kręgowców, u których mogą przejawiać się w tak spektakularnych zjawiskach jak odtworzenie całych kończyn po ich amputacji. U wyższych kręgowców zdolności regeneracyjne są znacznie bardziej ograniczone. Rozwój metod umożliwiających badanie globalnych profili ekspresji i metylacji...
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Characterization of a single-stranded DNA-binding-like Protein from Nanoarchaeum equitans - a nucleic acid binding protein with broad substrate specificity
PublikacjaBackground SSB (single-stranded DNA-binding) proteins play an essential role in all living cells and viruses, as they are involved in processes connected with ssDNA metabolism. There has recently been an increasing interest in SSBs, since they can be applied in molecular biology techniques and analytical methods. Nanoarchaeum equitans, the only known representative of Archaea phylum Nanoarchaeota, is a hyperthermophilic, nanosized,...
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Characterisation of Escherichia coli isolates from the blood of haematological adult patients with bacteraemia: translocation from gut to blood requires the cooperation of multiple virulence factors
PublikacjaThe aim of the study was to investigate whether there are unique pathotypes of Escherichia coli capable of transmission from the gastrointestinal tract to the vascular bed. The study included E. coli strains isolated from clinical materials collected from 115 patients suffering from haematologic malignancies diagnosed with bacteraemia. The genotyping techniques established that 89 E. coli isolates from the blood had the same genotype...
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A novel method of Mycobacterium tuberculosis complex strain differentiation using polymorphic GC-rich gene sequences.
PublikacjaTuberculosis 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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A new Toxoplasma gondii chimeric antigen containing fragments of SAG2, GRA1, and ROP1 proteins—impact of immunodominant sequences size on its diagnostic usefulness
PublikacjaThis study presents the first evaluation of new Toxoplasma gondii recombinant chimeric antigens containing three immunodominant regions of SAG2, GRA1, and one of two ROP1 fragments differing in length for the serodiagnosis of human toxoplasmosis. The recombinant chimeric antigens SAG2-GRA1-ROP1L (with large fragment of ROP1, 85-396 amino acid residues) and SAG2-GRA1-ROP1S (with a small fragment of ROP1, 85-250 amino acid residues)...
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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.
Rok 2014
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The use of a one-step PCR method for the identification of Microsporum canis and Trichophyton mentagrophytes infection of pets
PublikacjaIntroduction: 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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The optimal mixture of Toxoplasma gondii recombinant antigens (GRA1, P22, ROP1) for diagnosis of ovine toxoplasmosis
PublikacjaToxoplasmosis, caused by Toxoplasma gondii, is the major parasitic disease affecting sheep. Infection not only results in significant reproductive losses in these animals, but has public health implications since consumption of infected meat can facilitate zoonotic transmission. Although several serological tests are currently used for diagnosis of ovine toxoplasmosis, production of reliable reagents is a constraint and therefore...
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Structure-property relationships disclosed in the family of acridinium esters – chemiluminogenic molecules of great practical importance
PublikacjaAcridinium esters (AE), since their introduction to the analytics 30 years ago, make one of the most important groups of chemiluminogenic (CL) systems, that have been employed in medical diagnostics and environmental analysis today. So far, however, no systematic studies on the structureproperty relationships were carried out in this family of compounds. The latter features likely determine their practical utility and facilitate...
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ROLA SYGNALIZACJI KWASEM RETINOWYM W PROCESACH REGENERACJI I ROZWOJU ORGANIZMU
PublikacjaKwas retinowy jest cząsteczką sygnalną, odgrywającą kluczową rolę w trakcie rozwoju organizmu, która aktywuje różnicowanie wielu typów komórek. Zaburzenie sygnalizacji kwasem retinowym w okresie prenatalnym jest związany z poważnymi zaburzeniami rozwojowymi płodu, które mogą skutkować jego śmiercią. Wykazano także pozytywny wpływ sygnalizacji kwasem retinowym na zdolność do regeneracji uszkodzonych kończyn i tkanek u niektórych...
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Ogólna charakterystyka biofilmu drożdży z rodzaju Candida
PublikacjaDrożdże z rodzaju Candida to naturalne symbionty, które w przypadku obniżenia odporności organizmu mogą powodować grzybice. Właściwości chorobotwórcze związane są także z czynnikami wirulencji danego patogena. Oporność mikroorganizmów na leki stanowi poważny problem współczesnej medycyny. Grzyby posiadają wiele mechanizmów warunkujących lekooporność. Jednym z nich jest wytwarzanie biofilmu. Na powstawanie i rozwój biofilmu w znacznym...
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NASBA – IZOTERMICZNA AMPLIFIKACJA RNA
PublikacjaNASBA (ang. Nucleic Acid Sequence-Based Amplification) jest skuteczną i czułą techniką izotermicznej amplifikacji opartej na transkrypcji, służąca głównie do detekcji cząsteczek RNA. Znalazła ona zastosowanie do wykrywania RNA wirusowego, bakteryjnego i grzybiczego w próbkach klinicznych oraz żywości.
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Inhibition of amyloid fibril formation of hen egg white lysozyme by trimethylamine N-oxide at low pH
PublikacjaIn vitro inhibition of the formation of fibrous aggregates of proteins (amyloids) has gained increasing attention due to the number of diseases associated with protein misfolding and fibrillation. An interesting group of compounds for which pronounced activity against this phenomenon can be expected consists of low molecular weight substances (osmolytes) which have the ability to change protein stability. Here we investigate the...
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Identification and characterization of single-stranded DNA-binding protein from the facultative psychrophilic bacteria Pseudoalteromonas haloplanktis
PublikacjaSingle-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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Evaluation of possibilities in identification and susceptibility testing for Candida glabrata clinical isolates with the Integral System Yeast Plus (ISYP)
PublikacjaThe aim of this study was to evaluate possibilities of correct identification and susceptibility testing of C. glabrata clinical isolates with Integral System Yeast Plus (ISYP). For species identification, as the reference method, API Candida test and species-specific PCR reactions were used. The potential of antifungal susceptibility testing by the ISYP test was compared with the Sensititre Yeast One. Whilst the reference methods...
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ECHINOKANDYNY - MECHANIZM DZIAŁANIA ORAZ WYSTĘPOWANIE OPORNOŚCI U GRZYBÓW Z RODZAJU CANDIDA
PublikacjaEchinokandyny to lipopeptydowe leki przeciwgrzybicze, które są inhibitorami syntazy β-1,3-D-glukanowej. W praktyce klinicznej wykazano ich skuteczność wobec pleśni oraz grzybów drożdżopodobnych oraz minimalne efekty uboczne. Jednakże w ostatnich latach obserwuje się wzrastającą oporność na echinokandyny wśród klinicznych izolatów Candida spp. Przede wszystkim jest ona powiązana z występowaniem mutacji w obszarze genów fks, kodujących...
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Conserved motifs of MutL proteins
PublikacjatThe 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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Characterization of single-stranded DNA-binding proteins from the psychrophilic bacteria Desulfotalea psychrophila, Flavobacterium psychrophilum, Psychrobacter arcticus, Psychrobacter cryohalolentis, Psychromonas ingrahamii, Psychroflexus torquis, and Photobacterium profundum
PublikacjaBackground: Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination and repair in Bacteria, Archaea and Eukarya. In recent years, there has been an increasing interest in SSBs, since they find numerous applications in diverse molecular biology and analytical methods. Results: We report the characterization of single-stranded DNA-binding proteins from the psychrophilic bacteria Desulfotalea...
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BUDOWA I AKTYWNOŚĆ BIAŁEK NALEŻĄCYCH DO RODZINY TET
PublikacjaBiałka należące do rodziny TET (Tet1, Tet2, Tet3) są enzymami wykazującymi aktywność względem 5-metylocytozyny, która jest ważnym markerem epigenetycznym odgrywającym kluczową rolę w trakcie rozwoju organizmu i mającym istotny wpływ na poziom ekspresji genów. Białka TET przekształcają 5-metylocytozynę do 5-hydroksymetylocytozyny, 5-formylocytozyny i 5-karboksylocytozyny w trakcie następujących po sobie trzech reakcji utleniania....
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Budowa biofilmu chorobotwórczych drożdży i jego wpływ na oporność
PublikacjaBiofilm jest najpowszechniej występującą formą wzrostu mikroorganizmów w naturze. Ze względu na wysoką oporność na antybiotyki, jest on istotnym czynnikiem przewlekłych i nawracających zakażeń grzybiczych. Struktura biofilmu umożliwia dostarczenie składników odżywczych do głębszych warstw komórek i odprowadzenie metabolitów. Macierz zewnątrzkomórkowa pełni funkcję ochronne przed niekorzystnymi warunkami środowiska, a także utrudnia...
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Bacterial consortium an axenic cultures isolated from activated sewage sludge for biodegradation of imidazolium-based ionic liquid
PublikacjaExtensive research and increasing number of potential industrial applications made ionic liquids (ILs) important materials in design of new, cleaner technologies. Together with the technological applicability, the environmental fate of these chemicals is considered and significant efforts are being made in designing strategies to mitigate their potential negative impacts. Many ILs are proven to be poorly biodegradable and relatively...
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Assembly of Lipopolysaccharide in Escherichia coli Requires the Essential LapB Heat Shock Protein
PublikacjaHere, we describe two new heat shock proteins involved in the assembly of LPS in Escherichia coli, LapA and LapB (lipopolysaccharide assembly protein A and B). lapB mutants were identified based on an increased envelope stress response. Envelope stress responsive pathways control key steps in LPS biogenesis and respond to defects in the LPS assembly. Accordingly, the LPS content in ΔlapB or Δ(lapA lapB) mutants was elevated, with...
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A novel cold-active β-D-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB - gene cloning, purification and characterization
PublikacjaA gene encoding a novel β-D-galactosidase from the psychrotolerant Antarctic bacterium Arthrobacter sp. 32cB was isolated, cloned and expressed in Escherichia coli. The active form of recombinant β-D-galactosidase consists of two subunits with a combined molecular weight of approximately 257 kDa. The enzyme's maximum activity towards o-nitrophenyl-β-D-galactopyranoside was determined as occurring at 28 °C and pH 8.0. However, it...
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A new human IgG avidity test, using mixtures of recombinant antigens (rROP1, rSAG2, rGRA6), for the diagnosis of difficult-to-identify phases of toxoplasmosis
PublikacjaThe preliminary diagnostic utility of two mixtures of Toxoplasma gondii recombinant antigens (rROP1+rSAG2 and rROP1+rGRA6) in IgG ELISA and IgG avidity test has been evaluated. A total of 173 serum samples from patients with toxoplasmosis and seronegative people were examined. The sensitivity of IgG ELISA for rROP1+rSAG2 and rROP1+rGRA6 was 91.1% and 76.7%, respectively, while the reactivity for sera from patients where acute toxoplasmosis...
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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
PublikacjaWe 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...
Rok 2013
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Zakażenia o etiologii Candida spp. - epidemiologia, leczenie, diagnostyka
PublikacjaInfekcje wywoływane przez grzyby z rodzaju Candida są poważnym problemem klinicznym. LIczba zakażeń szpitalnych spowodowanych przez te drobnoustroje wzrosła dziesięciokrotnie w przeciągu ostatnich 20 lat. Spowodowane jest to poprzez coraz częstsze wystawianie populacji na działanie czynników spprzyjających rozwojowi tego typu infekcji. Wśród nich znajduje się leczenie: antybiotykami, steroidami, cytostatykami oraz lekami immunosupresyjnymi....
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Wytyczne dotyczące diagnostyki inwazyjnych zakażeń grzybiczych u pacjentów hematologicznych i onkologicznych
PublikacjaInwazyjne zakażenia grzybicze (ang. invasive fungal diseases - IFD) są główną przyczyną zachorowań i śmiertelności, szczególnie u pacjentów z nowotworami hematologicznymi. Grzyby, które są najczęstszymi przyczynami schorzeń u tego typu pacjentów to Aspergillus spp. oraz Candida albicans. Gatunki Candida nie-albicans oraz rosnąca liczba innych grzybów (np. Zygomycetes, Trichosporon, Fusarium spp.) są także coraz częściej wykrywane....
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Wybrane metody badania poziomu metylacji DNA w komórkach eukariotycznych
PublikacjaMetylacja regionów promotorowych jest jednym z głównych mechanizmów regulacji transkrypcji genów u eukariota. W komórkach ssaczych metylacja DNA zachodzi głównie na reszcie cytozyny występującej w dinukleotydach CpG i jest katalizowana przez metylotransferazy DNA (DNMT1, DNMT3A i DNMT3B) (Turek Plewa, Jagodziński 2005). Mapowanie 5 metylocytozyny w DNA genomowym jest jednym z narzędzi do badania wpływu metylacji na tkankowo-specyficzną...
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Toxoplasma gondii recombinant antigens as tools for serodiagnosis of human toxoplasmosis: current status of studies
PublikacjaToxoplasma gondii is a parasitic protozoan which is the cause of toxoplasmosis. Although human toxoplasmosis in healthy adults is usually asymptomatic, serious disease can occur in the case of congenital infections and immunocompromised individuals. Furthermore, despite the exact recognition of its etiology, it still presents a diagnostic problem. Diagnosis of toxoplasmosis is mainly based on the results of serological tests detecting...
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The usefulness of Toxoplasma gondii MIC1-MAG1-SAG1 chimeric antigen in the serodiagnosis of ovine toxoplasmosis
PublikacjaToxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii, which infects humans and most warm-blooded animals throughout the world. Although human toxoplasmosis in healthy adults is usually asymptomatic, a serious disease can occur in the case of congenital infection and immunocompromised individuals. Among food animals, sheep, along with goats and pigs, possess the highest incidence of T. gondii cysts in meat,...
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The role of epigenetics in regeneration
PublikacjaComplex changes in chromatin structure and at the transcriptional level occur from the creation of a single parental gamete throughout fertilization, embryo development and the life of an adult organism. Epigenetic changes, such as methylation and hydroxymethylation of DNA or histone methylation and acetylation, are an important part of these processes. Epigenetic regulation has an essential influence on gene expression level. DNA...
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THE GENE CLONING, OVEREXPRESSION, PURIFICATION AND BIOCHEMICAL CHARACTERISATION OF A NEW COLD-ADAPTED β–GALACTOSIDASE FROM ARTHROBACTER SP. VII-4
PublikacjaINTRODUCTION. β-Galactosidase [EC 3.2.1.23] is an enzyme that catalyzes the hydrolysis of O-glycosidic linkages in galactosides. It is commercially used in dairy industry for the production of milk with reduced lactose content. Potentially, the best method for lactose removal under cooling conditions should be carried out with a cold-adapted enzyme. AIM. The aim of this study was to determine the taxonomic affiliation of the isolate...
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Różnicowanie C. albicans i C. dubliniensis w oparciu o polimorfizm genu syntazy homocytrynianowej z wykorzystaniem metody PCR RFLP
PublikacjaNajczęstszym czynnikiem etiologicznym zakażeń wywołanych przez drożdżaki z rodzaju Candida jest C. albicans. Gatunek ten posiada bardzo zbliżone cechy fenotypowe do C. dubliniensis co często wiąże się z błędną identyfikacją. Skutkuje to wtórnym zakażeniem wywołanym przez współwystepujący gatunek C. dubliniensis, który nabywa oporność na flikonazol stosowany w terapii C. albicans. Celem badań była konstrukcja układu diagnostycznego...
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Recombinant !ermostable AP Exonuclease from Thermoanaerobacter tengcongensis: Cloning, Expression, Purification, Properties and PCR Application
PublikacjaApurinic/apyrimidinic (AP) sites in DNA are considered to be highly mutagenic and must be corrected to preserve genetic integrity, especially at high temperatures. !e gene encoding a homologue of AP exonuclease was cloned from the thermophilic anaerobic bacterium Thermoanaerobacter tengcongensis and transformed into Escherichia coli. The protein product showed high identity (80%) to human Ape1 nuclease, whereas to E. coli exonuclease...
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Phase transition of a dimorphic fungus Penicillium marneffei
PublikacjaPenicillium marneffei is a pathogenic fungus that afflicts immunocompromised individuals who live or travel in Southeast Asia. This species is unique, because it is the only dimorphic member of the genus Penicillium. Dimorphism results from a process, termed phase transition, which is regulated by temperature of incubation. At room temperature, the fungus grows filamentously (mould phase), but at body temperature (37°C), a uninucleate...
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PCR test for Microsporum canis identification
PublikacjaMicrosporum canis, for which the natural hosts are cats and dogs, is the most prevalent zoophilic agent causing tinea capitis and tinea corporis in humans. We present here a diagnostic PCR test for M. canis, since its detection and species identification is relevant to the choice of treatment and to the understanding of a probable source of infection. An M. canis-specific PCR was evaluated using 130 clinical isolates of dermatophytes...
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Novel primosomal protein B from Clostridium pastuerianum
PublikacjaPriB is a primosomal protein that catalyzes DNA replication in Procaryota. The replication pathway starts with PriA protein - the initiator protein that binds to a DNA replication fork, unwinds double-stranded DNA and role of PriB is to stabilize PriA on the DNA. However there are many biochemical differences in replication mechanism in bacteria and only some of them use PriB proteins. A few of PriB proteins were published and...
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Novel primosomal protein B from Clostridium pasteurianum
PublikacjaPriB is a primosomal protein that catalyzes DNA replication in Procaryota. The replication pathway starts with PriA protein - the initiator protein that binds to a DNA replication fork, unwinds double-stranded DNA and role of PriB is to stabilize PriA on the DNA. However there are many biochemical differences in replication mechanism in bacteria and only some of them use PriB proteins. A few of PriB proteins were published and...
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New PCR test for detection of Candida glabrata based on the molecular target chosen by the RAPD technique
PublikacjaRapid, reliable diagnosis is a necessary condition for the successful treatment of infections. Such diagnostic assays are continually being developed. #e paper presents a method for selecting the molecular target for PCR-based diagnostics based on the comparison of RAPD patterns. A sequence encoding Candida glabrata CBS138 hypothetical protein was selected. The limit of detection for PCR and real-time PCR reactions with DNA extracted...
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Mysz MRL - nowy model regeneracji
PublikacjaRegeneracja jest to zdolność do odtwarzania utraconych lub uszkodzonych kończyn, narządów i tkanek. Regeneracja wydaje się zdolnością niezbędną do życia, ale możliwości regeneracyjne są bardzo zróżnicowane w zależności od organizmu..Podczas gdy robaki płaskie są w stanie odtworzyć cały organizm z dowolnego jeog fragmentu, płazy regenerują utracone kończyny, przecięty.rdzeń kręgowy i poważne uszkodzenia serca, to zdolnosci regeneracyjne...
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MutL protein as a constituent of vsp, ner and mmr repair systems
PublikacjaMutS and MutL proteins are renowned mostly for their functions in well-characterized, post-DNA replication mis- match repair system (MMR). However, there is growing evidence that MMR system is not the only field of action for these pro- teins. Moreover, the participation in MMR does not even have to be their primary function. There are some reports indicat- ing involvement of MutL in BER, NER and VSP (very short patch repair)....
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Molecular and structural basis of inner core lipopolysaccharide alterations in Escherichia coli: incorporation of glucuronic acid and phosphoethanolamine in the heptose region.
PublikacjaIt is well established that lipopolysaccharide (LPS) often carries nonstoichiometric substitutions in lipid A and in the inner core. In this work, the molecular basis of inner core alterations and their physiological significance are addressed. A new inner core modification of LPS is described, which arises due to the addition of glucuronic acid on the third heptose with a concomitant loss of phosphate on the second heptose. This...
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Metylotransferazy ssaków – inhibitory i aktywatory
PublikacjaWśród metylotransferaz ssaków możemy wyróżnić metylotransferazy histonowe oraz metylotransferazy DNA. Metylacja DNA w obrębie wysp CpG oraz metylacja histonów wpływają na poziom ekspresji genów w komórkach ssaczych. Dodatkowo dzięki metylacji zapewniana jest stabilność i integralność genomu ludzkiego. Metylotransferazy DNA, będące głównym tematem tego artykułu, są często przedmiotem badań, gdyż nieprawidłowa metylacja DNA jest...
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Loop-mediated isothermal amplification (LAMP) as a diagnostic tool in detection of infectious diseases
PublikacjaLoop-mediated isothermal amplification (LAMP) is a gene amplification method which amplifies DNA with high specificity and efficiency under isothermal conditions. Because of its rapidity and simplicity, it is a valuable diagnostic tool in the early detection and identification of infectious diseases. LAMP method is based on the use of a set of four to six specially designed primers spanning six to eight distinct sequences on the...
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Ligation Mediated PCR methods, their possibilities and limitations
PublikacjaThe application of molecular diagnostics for bacterial strain typing or identifying bacteria at the strain level is of particular importance at present. However, among the many genotyping methods that are currently available, no single one is universally ideal. In this article, we thus review the Ligation Mediated PCR group methods as prospects from which to choose. We discuss different strategies for selecting the amplified PCR...
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Leukemia and risk of recurrent Escherichia coli bacteremia: genotyping implicates E. coli translocation from the colon to the bloodstream.
PublikacjaIn patients with leukemia, the portal(s) and reasons for the persistence of an Escherichia coli recurrent bacteremia remain unclear. Adult Hematology Clinic (AHC) databases at the State Clinical Hospital in Gdańsk were reviewed to evaluate the frequency of E. coli bacteremia between 2002 and 2005. Blood and bowel E. coli strains were obtained and the genetic relatedness of the strains was analyzed. The rate of E. coli bacteremia...
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Insight into heart regeneration
PublikacjaThe mammalian heart is thought to be incapable of regeneration because of the limited ability of cardiomyocytes. to proliferate. However, it has been recently shown that the neonatal murine heart is able to restore its normal function after coronary artery ligation or even partial resection. Unfortunately, this feature is lost in a few days after birth, although lower vertebrates as zebrafish or newt, retain this ability until adulthood. Therefore,...
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Identyfikacja sekwencji markerowych w mikologicznej diagnostyce molekularnej
PublikacjaProcedury diagnostyki mikologicznej coraz częściej wykorzystują metody molekularne, oparte na analizie kwasów nukleinowych. Pozwala to na skrócenie czasu analizy oraz polepszenie wielu innych, istotnych dla procesu diagnostycznego parametrów takich jak: czułość, powtarzalność, specyficzność oraz potencjał różnicujący. Istotnym etapem przy projektowaniu układów do diagnostyki molekularnej grzybów jest wybór celu molekularnego. Powszechnie...
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Fungal Typin Methods
PublikacjaThe 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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Epigenetic Basis of Regeneration: Analysis of Genomic DNA Methylation Profiles in the MRL/MpJ Mouse
PublikacjaEpigenetic regulation plays essential role in cell differentiation and dedifferentiation, which are the intrinsic processes involved in regeneration. To investigate the epigenetic basis of regeneration capacity, we choose DNA methylation as one of the most important epigenetic mechanisms and the MRL/MpJ mouse as a model of mammalian regeneration known to exhibit enhanced regeneration response in different organs. We report the...
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Epidemiological study of Toxoplasma gondii infection among cattle in Northern Poland
PublikacjaToxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. Because of medical and veterinary importance it is essential to study the prevalence of T. gondii infection among human and animals in various parts of the word. In this study, 4033 cattle from eight provinces of Northern Poland (belonging to 190 herds) were tested for IgG antibodies against T. gondii by an in-house ELISA technique based...
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Diagnostyka toksoplazmozy u kobiety ciężarnej, płodu i noworodka — stan obecny i nowe możliwości
PublikacjaW artykule opisano problem toksoplazmozy, skutki oraz obecną diagnostykę. Przedstawiono metody serologiczne pozwalające wykryć pierwotne zarażenie T. gondii u kobiet ciężarnych oraz szereg technik pośrednich i bezpośrednich wykorzystywanych w celu potwierdzenia bądź wykluczenia toksoplazmozy wrodzonej u noworodka. Ponadto wskazano potencjalne możliwości w diagnostyce toksoplazmozy, jakie niosą za sobą nowe techniki (np. 2D Western-blotting),...
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Cold-adapted bacterial lipolytic enzymes and their applications
PublikacjaLipolytic enzymes, esterases and lipases catalyze the hydrolysis and transesterification of fatty acid esters. This class of enzymes is the most important group of biocatalysts for biotechnological applications. From this perspective, coldactive lipolytic enzymes from cold adapted bacteria display attractive properties for some industrial purposes, including high catalytic activity at low temperatures and low thermostability. To...
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Cloning and characterization of a novel cold-active glycoside hydrolase family 1 enzyme with beta-glucosidase, beta-fucosidase and beta-galactosidase activities.
PublikacjaBackground: Cold-active enzymes, sourced from cold-adapted organisms, are characterized by high catalytic efficiencies at low temperatures compared with their mesophilic counterparts, which have poor activity. This property makes them advantageous for biotechnology applications as it: (i) saves energy costs, (ii) shortens the times for processes operated at low temperatures, (iii) protects thermosensitive substrates or products...
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Characterization of single-stranded DNA-binding proteins from the psychrophilic bacteria
PublikacjaSingle-stranded DNA-binding proteins (SSBs) are indispensable elements in the cells of all living organisms. SSB proteins interact with ssDNA insequence in an independent manner, preventing them from forming secondary structures and from degradation by nucleases. In this way, SSB-binding proteins participate in all processes involving ssDNA, such as replication, repair and recombination. Although there are differences in amino...
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Characterization of single-stranded DNA-binding proteins from the psychrophilic bacteria
PublikacjaSingle-stranded DNA-binding proteins (SSBs) are indispensable elements in the cells of all living organisms. SSB proteins interact with ssDNAinsequence in an independent manner, preventing them from forming secondary structures and from degradation by nucleases. In this way, SSB-binding proteins participate in all processes involving ssDNA, such as replication, repair and recombination.Although there are differences in amino acid...
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BIODIVERSITY OF YEASTS ISOLATED FROM CAVES OF THE POLISH TATRA MOUNTAINS, AND ITS BIOTECHNOLOGICAL POTENTIAL
PublikacjaMost of the Earth’s biosphere is permanently or periodically exposed to temperatures below 5°C. Cold habitats harbor a wide diversity of psychrophiles. For many years Antarctica has been the geographic area preferred for studying the diversity and properties of cold-adapted microorganisms. However, the non-Antarctic habitats like Himalayas or Alps are also extensively explored. The Tatra Mountains are Europe’s second-highest massif...
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Biochemical characteristic of biofilm of uropathogenic Escherichia coli Dr+ strains.
PublikacjaUrinary tract infections caused by Escherichia coli are very common health problem in the developed countries. The virulence of the uropathogenic E. coli Dr+ IH11128 is determined by Dr fimbriae, which are homopolymeric structures composed of DraE subunits with the DraD protein capping the fiber. In this study, we have analyzed the structural and biochemical properties of biofilms developed by E. coli strains expressing Dr fimbriae...
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Białka prokariotycznego systemu MMR: rola oraz interakcje
PublikacjaUszkodzenia komórkowego DNA w postaci błędnie sparowanych lub niesparowanych zasad azotowych są wynikiem niewłaściwie zachodzących procesów metabolizmu DNA, takich jak replikacja, rekombinacja oraz naprawa DNA. Utrwalenie tego rodzaju zmian prowadzi do powstania mutacji. System MMR (ang. DNA mismatch repair) usuwa te uszkodzenia przyczyniając się do zwiększenia wierności replikacji DNA oraz utrzymania stabilności genomu komórki....
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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
PublikacjaINTRODUCTION: 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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Beta-Galactosidases of Pseudoalteromonas sp. – Characteristics and application for lactose hydrolysis and galactopyranosides synthesis
PublikacjaBeta-galactosidase [EC 3.2.1.23] is an enzyme that catalyzes the hydrolysis of O-glycosidic linkages in galactosides. It has biotechnological applications in the dairy industry for the production of milk with a low content of lactose and galactooligosacharides for use in healthy food. The transgalactosylation activity of beta-galactosidase can be used in the synthesis of alkyl galactopyranosides. Due to the fact that conditions of...
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Beta-galactosidases from a psychrotolerant Arthrobacter isolates and their potential use
PublikacjaEnzymes from extremophiles, especially psychrophiles and psychrotolerant microorganisms are currently intensively studied because of their unusual structural and functional properties. One of these enzymes is a cold-adapted - galactosidase (EC 3.2.1.23) which catalyzes the hydrolysis of - 1,4-glycosidic bonds in -galactosides. The most common - galactoside is lactose, a disaccharide which makes up 2-7% of milk. The majority of...
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A subset of two adherence systems, acute pro-inflammatory pap genes and invasion coding dra, fim, or sfa, increases the risk of Escherichia coli translocation to the bloodstream
PublikacjaAn analysis of the phylogenetic distribution and virulence genes of Escherichia coli isolates which predispose this bacteria to translocate from the urinary tract to the bloodstream is presented. One-dimensional analysis indicated that the occurrence of P fimbriae and α-hemolysin coding genes is more frequent among the E. coli which cause bacteremia. However, a two-dimensional analysis revealed that a combination of genes coding...
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A study on the interaction of rhodamine B with methylthioadenosine phosphorylase protein sourced from an Antarctic soil metagenomic library.
PublikacjaThe presented study examines the phenomenon of the fluorescence under UV light excitation (312 nm) of E. coli cells expressing a novel metagenomic-derived putative methylthioadenosine phosphorylase gene, called rsfp, grown on LB agar supplemented with a fluorescent dye rhodamine B. For this purpose, an rsfp gene was cloned and expressed in an LMG194 E. coli strain using an arabinose promoter. The resulting RSFP protein was purified...
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A new division of bacterial UvrA homologues
PublikacjaThe 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...
Rok 2012
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Sick building syndrome due to molds
PublikacjaThe 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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Sequence variants of p21 gene of the MRL/MpJ mouse: a possible association with the deficit of p21 protein and regenerative phenotype.
PublikacjaThe MRL/MpJ mouse is known for its enhanced regeneration abilities that manifested themselves by scarless ear-hole wound closure. The deficit of p21 found in the MRL is associated with the regenerative capacity as the p21 gene knockout in a mouse strain unrelated to the MRL mouse results in an ear-hole closure effect, similar to that observed in the MRL mouse. Cdkn1a gene encodes p21 protein, a cyclin-dependent kinase inhibitor,...
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Real-Time PCR: molecular technique of many applications
PublikacjaReal-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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Reakcje krzyżowe w immunodiagnostyce
PublikacjaZaprezentowano problem reakcji krzyżowych w immunodiagnostyce.
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Production of Toxoplasma gondii MAG1 recombinant antigen in Leishmania tarentolae expression system
PublikacjaThe 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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Problem reakcji krzyżowych w immunodiagnostyce na przykładzie neosporozy i toksoplazmozy
PublikacjaW 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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Postępy w zakresie analizy powietrza mającej na celu identyfikację grzybów środowiskowych
PublikacjaPowietrze 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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Podłoże nieswoistego zapalenia jelit
PublikacjaZaprezentowano możliwe czynniki ryzyka wystąpienia nieswoistego zapalenia jelit.
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Pilicides effectively hinder chaperone-usher interactions in all Gram-negative bacteria
PublikacjaThe 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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Peptidoglycan hydrolases - potential weapons against Staphylococcus aureus
PublikacjaBacteria 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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PCR-ELISA: inexpensive alternative to quantitative PCR
PublikacjaPCR-ELISA (polymerase chain reaction-enzyme linked immunosorbent assay), a combination of PCR and ELISA methods, has been used since late 1980s. The technique is based on specially labelled DNA fragments which are captured by specific DNA sequences and detected by antibodies. The whole procedure of PCR-ELISA is divided into three steps: DNA extraction, PCR reaction and detection by ELISA. The method has been found as very specific...
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Nieswoiste zapalenie jelit
PublikacjaNieswoiste 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
PublikacjaThe 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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MIC1-MAG1-SAG1 chimeric protein, a most effective antigen for detection of human toxoplasmosis
PublikacjaThis study describes a Toxoplasma gondii IgG ELISA test based on a new chimeric antigen containing three immunodominant regions from MIC1, MAG1 and SAG1 proteins of a parasite and shows that this test is useful for diagnostic purposes and may replace the lysed and whole-cell antigens.
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Material Flow Analysis in a cooked mussel processing industry
PublikacjaGalicia (NW Spain) is an ideal environment for mussel culture owing to its excellent natural conditions. It is the first Spanish and European mussel producer and the second worldwide after China. More than 50% of the mussel collected in Galicia is used in the processing industry, which produces different products, as cooked mussel that is canned, frozen or used in the food industry. In this paper a methodology based on Life Cycle...
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Isolation and identification of Mycobacterium avium subspecies silvaticum from a horse
PublikacjaPo 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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Induction of immune responses in sheep by vaccination with liposome-entrapped DNA complexes encoding Toxoplasma gondii MIC3 gene
PublikacjaToxoplasma gondii is a parasite that has been extensively studied due to its medical and veterinary importance in terminating pregnancies. Consequently, a satisfactory vaccine is required to control itsadverse effects on pregnant animals. The microneme protein, MIC3, is a major adhesion protein that binds to the surface of host cells and parasites, and is therefore a potential vaccine against T. gondii.The viability of MIC3 as...
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Importance of biofilm formation by uropathogenic E. coli Dr+ strains in urinary tract infections
PublikacjaImportance of biofilm formation by uropathogenic E. coli Dr+ strains in urinary tract infections
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Immunization with an anti-idiotypic antibody against the broadly lipopolysaccharide-reactive antibody WN1 222-5 induces Escherichia coli R3-core-type specific antibodies in rabbits
PublikacjaThe mouse monoclonal antibody (mAb) WN1 222-5 recognizes a carbohydrate epitope in the inner core region of LPS that is shared by all strains of Escherichia coli and Salmonella enterica and is able to neutralize their endotoxic activity in vitro and in vivo. Immunization of mice with mAb WN1 222-5 yielded several anti-idiotypic mAbs one of which (mAb S81-19) competitively inhibited binding of mAb WN1 222-5 to E. coli and Salmonella...