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Search results for: recombinant protein
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Immunogenic and protective activity of recombinant chimeric Toxoplasma gondii proteins
PublicationToxoplasmosis, one of the most common parasitic invasions worldwide, may pose a great threat for individuals with weakened immune system. The parasite also causes considerable economic losses due to infection of livestock. To date there is no efficient immunoprophylaxis for humans and animals, which would provide long-lasting protection against T. gondii invasion. Chimeric proteins, containing several selected fragments of parasite...
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Hyperosmolarity adversely impacts recombinant protein synthesis by Yarrowia lipolytica—molecular background revealed by quantitative proteomics
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Toxoplasma gondii Recombinant antigen AMA1: Diagnostic Utility of Protein Fragments for the Detection of IgG and IgM Antibodies
PublicationToxoplasma gondii is an important zoonotic protozoan that infects a wide variety of vertebrates as intermediate hosts. For this reason, the diagnosis of this disease is very important and requires continuous improvement. One possibility is to use recombinant antigens in serological tests. Apical membrane antigen 1 (AMA1), a protein located in specific secretory organelles (micronemes) of T. gondii, is very interesting in regard...
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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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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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Function and Frustration of Multi-Drug ABC Exporter Protein and Design of Model Proteins for Drug Delivery Using Protein Hydration Thermodynamics
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Novel monovalent and multivalent recombinant proteins of Borrelia burgdorferi sensu lato with potential diagnostic value – construction and biotechnological production
PublicationThe diverse antigenic structure of Borrelia burgdorferi sensu lato (s.l.) and the low degree of protein sequence conservation between genospecies causes many limitations in serodiagnosis of Lyme disease (LD). Using expression systems based on Escherichia coli, five monovalent B. burgdorferi s.l. recombinant proteins were produced. i.e., BB0108, BB0126, BB0298, BB0323, BB0689 (each in three variants derived from Borrelia afzelii,...
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Elastic-contractile model proteins: Physical chemistry, protein function and drug design and delivery
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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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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
PublicationIntroduction 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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Toxoplasma gondii: Enzyme-Linked Immunosorbent Assay using different fragments of recombinant microneme protein 1 (MIC1) for detection of immunoglobulin G antibodies
PublicationTrzy rekombinantowe fragmenty białka antygenowego MIC1 Toxoplasma gondii (r-MIC1ex2, r-MIC1ex34 i r-MIC1) zostały wyprodukowane jako białka fuzyjne (zawierające dwie domeny oligohistydynowe na N- i C-końcu) w komórkach bakteryjnych Escherichia coli. Homogenne preparaty antygenów rekombinantowych otrzymane z zastosowaniem jednoetapowego oczyszczania metodą chromatografii metalopowinowactwa, wykorzystano następnie do immunoidentyfikacji...
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How proteins bind to DNA: target discrimination and dynamic sequence search by the telomeric protein TRF1
PublicationTarget search as performed by DNA-binding proteins is a complex process, in which multiple factors contribute to both thermodynamic discrimination of the target sequence from overwhelmingly abundant off-target sites and kinetic acceleration of dynamic sequence interrogation. TRF1, the protein that binds to telomeric tandem repeats, faces an intriguing variant of the search problem where target sites are clustered within short fragments...
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Isolation and properties of recombinant trehalose synthase
PublicationBiosynthesis of trehalose is carried out via various matabolic pathways, including the use of trehalose synthase. Bacteria of the genus Deionococcus are microorganisms which produce trehalose synthase catalizing conversion of maltose into trehalose. In this work recombinant trehalose synthase from D.radiodurans was characterized.
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Extension of the Unres Package for Physics-Based Coarse-Grained Simulations of Proteins and Protein Complexes to Very Large Systems
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Characterization of recombinant homocitrate synthase from Candida albicans
PublicationLYS21 and LYS22 genes from Candida albicans encoding isoforms of homocitrate synthase (HCS), an enzyme catalyzing the first committed step in the L-lysine biosynthetic pathway, were cloned and expressed as NoligoHistagged fusion proteins in Escherichia coli. The purified gene products revealed HCS activity, i.e. catalyzed the condensation of α-ketoglutarate with acetyl-coenzyme A to yield homocitrate. The recombinant enzymes were purified...
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UNRES-Dock—protein–protein and peptide–protein docking by coarse-grained replica-exchange MD simulations
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The first study on the usefulness of recombinant tetravalent chimeric proteins containing fragments of SAG2, GRA1, ROP1 and AMA1 antigens in the detection of specific anti-Toxoplasma gondii antibodies in mouse and human sera
PublicationThis study presents an evaluation of four tetravalent recombinant chimeric proteins containing fragments of the Toxoplasma gondii antigens, SAG2, GRA1, ROP1 and AMA1, as potential replacements of a the soluble, whole-cell tachyzoite lysate (TLA) used in serological assays. Recombinant chimeric proteins (SAG2-GRA1-ROP1-AMA1N, AMA1N-SAG2-GRA1-ROP1, AMA1C-SAG2-GRA1-ROP1, and AMA1-SAG2-GRA1-ROP1) obtained by genetic engineering were...
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Use of MAG1 recombinant antigen for diagnosis of Toxoplasma gondii infection in humans
PublicationPraca opisuje klonowanie, oczyszczanie oraz zastosowanie w immunodiagnostyce toksoplazmozy antygenu rekombinantowego macierzy cyst tkankowych (MAG1) Toxoplasma gondii. Zastosowany system ekspresyjny pozwolił uzyskać dużą ilość rekombinantowego antygenu, który następnie wykorzystano w teście ELISA do wykrywania specyficzych przeciwciał anty-T. gondii klasy IgG w surowicach pacjentów chorych na toksoplazmozę. W przeprowadzonych badaniach...
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Toxoplasma gondii: Chimeric Dr fimbriae as a recombinant vaccine against toxoplasmosis
PublicationW publikacji przedstawiono protekcyjne i immunogenne właściowści chimerycznych fimbrii typu Dr zawierających sekwencje epitopowe antygenów SAG1, MAG1 oraz GRA1 Toxoplasma gondii. Uzyskane wyniki porównano z właściwościami immunogennymi antygenowych białek rekombinantowych oraz poliwalentnego antygenu natywnego. Antygeny rekombinantowe indukują silną odpowiedź immunologiczną i w 89% zapobiegają formowaniu się cyst w mózgu myszy....
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Toxoplasma gondii: an evaluation of diagnostic value of recombinant antigens in a murine model
PublicationPraca dotyczy oceny użyteczności sześciu rekombinantowych białek antygenowych Toxoplasma gondii (GRA1, GRA6, GRA7, p35, SAG1 i SAG2) w diagnozowaniu doświadczalnej toksoplazmozy u myszy. Przeprowadzone badania pozwoliły na określenie stopnia reaktywności zastosowanych antygenów z surowicami z wczesnej i przewlekłej toksoplazmozy. W przypadku ostrej fazy choroby obserwowano silną odpowiedź humoralną na antygeny GRA7, GRA6 i p35,...
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Toxoplasma gondii Recombinant Antigens in the Serodiagnosis of Toxoplasmosis in Domestic and Farm Animals
PublicationToxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, and is a parasitic disease that occurs in all warm-blooded animals, including humans. Toxoplasmosis is one of the most common parasitic diseases of animals and results in reproductive losses. Toxoplasmosis in humans is usually caused by eating raw or undercooked meat or consuming dairy products containing the parasite. Diagnosis of toxoplasmosis is currently...
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The Protein Folding Problem
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Influence of Osmolytes on Protein and Water Structure: A Step To Understanding the Mechanism of Protein Stabilization
PublicationResults concerning the thermostability of hen egg white lysozyme in aqueous solutions with stabilizing osmolytes, trimethylamine-N-oxide (TMAO), glycine (Gly), and its N-methyl derivatives, N-methylglycine (NMG), N,N-dimethylglycine (DMG), and N,N,N-trimethylglycine (betaine, TMG), have been presented. The combination of spectroscopic (IR) and calorimetric (DSC) data allowed us to establish a link between osmolytes’ influence on...
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Proteins
PublicationStruktura białek wpływa na ich wartość odżywczą, właściwości funkcjonalne wżywności oraz reakcje w surowcach i produktach żywnościowych. Warunki prze-chowywania oraz obróbki w przemyśle i w czasie przygotowywania kulinarnegowywołują przemiany białek istotne dla jakości żywności. Przez celowe, enzy-matyczne i chemiczne modyfikowanie składu i struktury białek można wpływaćna ich właściwości funkcjonalne w żywności i wartość...
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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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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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Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli
PublicationA trehalose synthase gene from Deinococcus radiodurans(DSMZ 20539)containing 1659 bp reading frame encoding 552 amino acids was amplified using PCR. The gene was finally ligated into pET30Ek/LIC vector and expressed after isopropyl β-d-thiogalactopyranoside induction in Escherichia coli (DE3) Rosetta pLysS. The recombinanttrehalose synthase (DraTreS) containing a His6-tag at the C-terminus was purified by metal affinity chromatography...
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Cloning, expression, purification and characterization of recombinant trehalose synthase from Deinococcus radiodurans.
PublicationTrehalose (α-D-glucopyranosyl-1,1-α-D-glucopyranoside) is a nonreducing disacharide in which the two glucose molecules are linked trough a α-1,1-glycosidic bond. Trehalose is readily hydrolyzed to glucose and can be used as a reserve of that sugar in the cell. The presence of trehalose was found in the cells of fungi and yeasts, bacteria, nematodes, insects, eggs, pupae and some plants. The characteristics of trehalose make it...
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Efficient production of Staphylococcus simulans lysostaphin in a benchtop bioreactor by recombinant Escherichia coli.
PublicationLysostaphin is an enzyme with bactericidal activity against Staphylococcus aureus and other staphylococcal species. In spite of many advantages and promising results of preliminary research, the enzyme is still not widely used in medicine, veterinary or as a food preservative. One of the most important factors limiting application of the enzyme in clinical or technological practice is the high costs of its production. In the present...
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Testing the Diagnostic Utility of Recombinant Toxoplasma Gondii Chimeric Antigens – Generated Datasets
PublicationThe datasets titled Toxoplasma gondii recombinant chimeric antigens – IgM and IgG ELISAs – mouse serum samples and Toxoplasma gondii recombinant chimeric antigens – IgG and IgM ELISAs – human serum samples contain absorbance measurements obtained during serological tests using mouse and human sera in enzyme-linked immunosorbent assay (ELISA) tests based on recombinant chimeric antigens. The datasets allows a comparison of absorbance...
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A method for the production of D-tagatose using a recombinant Pichia pastoris strain secreting beta-D-galactosidase from Arthrobacter chlorophenolicus and a recombinant L-arabinose isomerase from Arthrobacter sp. 22c
PublicationD-Tagatose is a natural monosaccharide which can be used as a low-calorie sugar substitute in food, beverages and pharmaceutical products. It is also currently being tested as an anti-diabetic and obesity control drug. D-Tagatose is a rare sugar, but it can be manufactured by the chemical or enzymatic isomerization of D-galactose obtained by a beta-D-galactosidase-catalyzed hydrolysis of milk sugar lactose and the separation of...
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On the Origin of Protein Superfamilies and Superfolds
PublicationDistributions of protein families and folds in genomes are highly skewed, having a small number of prevalent superfamiles/superfolds and a large number of families/folds of a small size. Why are the distributions of protein families and folds skewed? Why are there only a limited number of protein families? Here, we employ an information theoretic approach to investigate the protein sequence-structure relationship that leads to...
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Colloidal aspects of protein digestion
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Stabilizing agents and protein stability
PublicationBiałka są kluczowymi elementami żywych organizmów, jak również efektywnymi katalizatorami w biotechnologii i medycynie. Szybki rozwój wielu gałęzi biotechnologii wiąże się z coraz większym zapotrzebowaniem na produkty białkowe charakteryzujące się wysoką stabilnością. Jednym ze sposobów osiągnięcia większej stabilności białek jest wykorzystanie małych związków organicznych, tzw. osmolitów. Do tej grupy związków należą aminokwasy,...
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Alternative and New Protein Sources
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The importance of the shape of the protein–water interface of a kinesin motor domain for dynamics of the surface atoms of the protein
PublicationA single kinesin motor domain immersed in water has been investigated using molecular dynamics. It has been found that local properties of water in the solvation shell change along with the nature of the neighboring protein surface. However, a detailed analysis leads to the conclusion that the geometrical features of hydrogen bonds and overall structure of kinesin hydration water are not very different from bulk water. The local...
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The importance of the shape of the protein-water interface of a kinesin motor domain for dynamics of the surface atoms of the protein
PublicationSingle kinesin motor domain immersed in water has been investigated using molecular dynamics. It has been found that local properties of water in solvation shell change along with the nature of neighboring protein surface. However, a detailed analysis leads to the conclusion that the geometrical features of hydrogen bonds and overall structure of kinesin hydration water is not very different from bulk water. The local values of...
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Endocrine responses following exhaustive strength exercise with and without the use of protein and protein-carbohydrate supplements
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Protein overproduction alters exosome secretion in Chinese hamster ovary cells
PublicationDespite the abundance of available cell lines, nearly 70% of all recombinant therapeutic proteins today are produced in Chi-nese hamster ovary (CHO) cells. The impact of protein overproduction on the secretion of exosomes by CHO cells has been investigated here. Increased secretion of extracellular vesicles (EVs) by protein overexpressing CHO cells was demonstrated with protein content assay, nanoparticle tracking analysis, and...
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Toxoplasma gondii recombinant antigens as tools for serodiagnosis of human toxoplasmosis: current status of studies
PublicationToxoplasma 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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Use of Escherichia coli Nissle 1917 producing recombinant colicins for treatment of IBD patients
PublicationPatients with Crohn’s Disease and ulcerative colitis infected with Adherent-Invasive Escherichia coli strains constitute the largest group among Inflammatory Bowel Disease subjects, when taking into account all known etiological agents of the disease. A possible link between these pathogenic bacteria and inflammation process has gained the confidence in recently published papers. Observed enteric neuroglial cells apoptosis and...
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Occlusion phenomenon of redox probe by protein as a way of voltammetric detection of non-electroactive C-reactive protein
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Chimeric Dr fimbriae with a Herpes Simplex virus type 1 epitope as a model for a recombinant vaccine.
PublicationBiałko adhezyny DraE uropatogennych szczepów E. coli Dr+ zostało wykorzystane jako nośnik 11a a determinanty antygenowej glikoproteiny D wirusa HSV ty-pu 1. Sekwencja kodująca epitop HSV została wklonowana do genu draE w obrębie regionu N- terminalnego eksponowanej powierzchniowo domeny 2. Ekspresjachimerycznych białek DraE-HSV została potwierdzona za pomocą mikroskopii immunofluorescencyjnej i elektronowej.
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Efficient production of the Toxoplasma gondii GRA6, p35 and SAG2 recombinant antigens and their applications in the serodiagnosis of toxoplasmosis
PublicationSkonstruowano system ekspresji rekombinantowych białek antygenowych GRA6, p35 i SAG2 Toxoplasma gondii. Uzyskano miligramowe ilości białek fuzyjnych po zastosowaniu jednoetapowego oczyszczania z wykorzystaniem metody chromatografii metalopowinowactwa. Przydatność otrzymanych antygenów do konstrukcji serologicznego testu diagnostycznego oszacowano z wykorzystaniem testu ELISA, w którym zastosowano 99 surowic pobranych od pacjentów...
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The optimal mixture of Toxoplasma gondii recombinant antigens (GRA1, P22, ROP1) for diagnosis of ovine toxoplasmosis
PublicationToxoplasmosis, 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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Iron–Sulfur Cluster Biogenesis Chaperones: Evidence for Emergence of Mutational Robustness of a Highly Specific Protein–Protein Interaction
PublicationBiogenesis of iron–sulfur clusters (FeS) is a highly conserved process involving Hsp70 and J-protein chaperones. However, Hsp70 specialization differs among species. In most eukaryotes, including Schizosaccharomyces pombe, FeS biogenesis involves interaction between the J-protein Jac1 and the multifunctional Hsp70 Ssc1. But, in Saccharomyces cerevisiae and closely related species, Jac1 interacts with the specialized Hsp70 Ssq1,...
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The Role of Proteins in Food
PublicationThis chapter describes the effect of proteins on the sensory attributes and the biological value and safety of foods. The role of proteins depends on their amino acid composition and structure, on changes due to storage and processing, as well as on interactions with other food components. The effect on the sensory quality of foods is brought about by hydrophobicity, solubility, water holding capacity, gelling, film formation,...
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New recombinant chimeric antigens, P35-MAG1, MIC1-ROP1, and MAG1-ROP1, for the serodiagnosis of human toxoplasmosis
PublicationThe 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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Application of recombinant antigens in serodiagnosis of swine toxoplasmosis and prevalence of Toxoplasma gondii infection among pigs in Poland
PublicationNiniejsza praca przedstawia częstość występowania toksoplazmozy w populacji świń obejmującej 1754 osobników pochodzących z różnych regionów Polski, która określona została z wykorzystaniem testu IgG ELISA opartego na antygenie natywnym pasożyta. Specyficzne przeciwciała anty-T. gondii klasy IgG zostały wykryte u 19,2% zwierząt z testowanej grupy. Ponadto w pracy oszacowano użyteczność diagnostyczną w rozpoznawaniu toksoplazmozy...
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C-1305 inactivation of recombinant human cytochrome P450, CYP1A2 and CYP3A4, is due to mechanism-based inhibition
PublicationPodjęto próbę określenia mechanizmu obserwowanej inhibicji izoenzymów CYP1A2 i CYP3A4 przez pochodną triazoloakrydonu C-1305.W pierwszym kroku stwierdzono, że hamowanie aktywnosci obu enzymów jest procesem nieodwracalnym. W kolejnych etapach badań wykazano między innymi, że obecność NADPH zwiększa istotnie stopień obserwowanej inhibicji, co wskazało, że mamy do czynienia z inhibicją samobójczą opartą na mechanizmie działania (ang....