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Search results for: DNA REPAIR ENZYMES
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MutS3: a MutS homologue of unknown biological function
PublicationThe homologues of MutS proteins are widespread among both Prokaryotes and Eukaryotes. MutS designated as MutS1 is a part of MMR (mismatch repair) system which is responsible for removal of mispaired bases and small insertion/deletion loops in DNA. Initially, the only MutS homologues known were those engaged in mismatch repair and these were later designated as MutS1. Subsequently, the MutS2 homologue was distinguished. MutS2 does...
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Możliwości zastosowań, potencjalne źródła oraz ewolucja technologiczna sposobu otrzymywania trehalozy ze szczególnym uwzględnieniem enzymu syntazy trehalozy [EC 5.4.99.16]
PublicationJedna z wiodących gałęzi współczesnej biotechnologii opiera się na produkcji białek – przede wszystkim enzymów. Są one powszechnie wykorzystywane: od przemysłu chemicznego poprzez przemysł spożywczy aż po medycynę i życie codzienne (czy ktoś wyobraża sobie w dzisiejszych czasach proszek do prania „bez enzymu”). Przed rozwojem nowoczesnych metod biologii molekularnej i inżynierii genetycznej pozyskiwano interesujące substancje z...
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Identification and characterization of single-stranded DNA-binding protein from the facultative psychrophilic bacteria Pseudoalteromonas haloplanktis
PublicationSingle-stranded DNA-binding protein (SSB) plays an important role in DNA metabolism such as DNAreplication, repair, and recombination, and is essential for cell survival. This study reports on the ssb-likegene cloning, gene expression and characterization of a single-stranded DNA-binding protein of Pseudoal-teromonas haloplanktis (PhaSSB) and is the first report of such a protein from psychrophilic microorganism.PhaSSB possesses...
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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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Electrochemistry meets enzymes: Investigation of the biotransformation pathway of C-1311 based on electrochemical simulation in comparison to in vitro methods.
PublicationThe knowledge of the metabolic pathways and the biotransformation of new drugs is one of the major challenges in pharmaceutical research. It is crucial for elucidation of degradation routes of the new biologically active compounds, especially in the area of possible toxicity. Conventional in vitro drug metabolism studies are based on incubating drug candidate with e.g. hepatocytes or, most importantly, liver cell microsomes and...
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Molecular Targets for Anticandidal Chemotherapy
PublicationA relatively small number of anticandidal chemotherapeutics used in clinical practice is at least in part consequence of a limited number of their molecular targets: ergosterol in the membrane, lanosterol demethylase, b(1!3) glucan synthase, and DNA/RNA biosynthesis. Much more potential novel targets have been revealed by the comparative genomic studies identifying essential genes unique for Candida albicans or resulted from recognition...
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A new division of bacterial UvrA homologues
PublicationThe UvrA protein is a DNA-binding and damage-recognition enzyme which participates in the prokaryotic type nucleotide excision repair (NER) pathway. It has recently been noted that some bacterial genomes comprise additional uvrA genes which encode five distinct types of UvrA homologue. We investigated the sequences of over 2400 bacterial genomes and found 130 examples of bacteria containing uvrA 2 genes. The sequence analyses conducted...
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Biochemistry - Laboratory exercises - Nowy
e-Learning CoursesExerise 1. The ability to accumulate metal cations by edible mushrooms, comparison with edible plants Exercise 2. Measurement of activity of glutathione S-transferases - the detoxifying enzymes present in the cytosol of eukaryotic cells Exercise 3. Cell membranes permeability Exercise 4. Detection of mutagenic substances using Ames test Exercise 5. Gluten content in food products part 1. Exercise 6. Gluten content in food...
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Biochemistry - Laboratory exercises
e-Learning CoursesExerise 1. The ability to accumulate metal cations by edible mushrooms, comparison with edible plants Exercise 2. Measurement of activity of glutathione S-transferases - the detoxifying enzymes present in the cytosol of eukaryotic cells Exercise 3. Cell membranes permeability Exercise 4. Detection of mutagenic substances using Ames test Exercise 5. Gluten content in food products part 1. Exercise 6. Gluten content in food...
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A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification
PublicationCandida sp. have been responsible for an increasing number of infections, especially in patients with immunodeficiency. Species specific differentiation of the Candida sp. is difficult in routine diagnosis. This identification can have a highly significant association in therapy and prophylaxis. This work has shown a new application of terminal restriction fragment length polymorphism (t-RFLP) method in molecular identification...
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Epigenetic inhibitor zebularine activates ear pinna wound closure in the mouse
PublicationBackground:Most studies on regenerative medicine focus on cell-based therapies and transplantations.Small-molecule therapeutics, though proved effective in different medical conditions, have not been extensivelyinvestigated in regenerative research. It is known that healing potential decreases with development and devel-opmental changes are driven by epigenetic mechanisms, which suggests epigenetic repression of regenerativecapacity.Methods:We...
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Genetic and Epigenetic Aspects of Atopic Dermatitis
PublicationAtopic dermatitis is a heterogeneous disease, in which the pathogenesis is associated with mutations in genes encoding epidermal structural proteins, barrier enzymes, and their inhibitors; the role of genes regulating innate and adaptive immune responses and environmental factors inducing the disease is also noted. Recent studies point to the key role of epigenetic changes in the development of the disease. Epigenetic modifications...
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Activation of endogenous regenerative potential in mammals using epigenetic inhibitor zebularine
PublicationIn this work, a nucleoside inhibitor of DNA methyltransferases, zebularine, was investigated as a wound healing and regeneration promoting agent. It was found that a high dose of intraperitoneally delivered zebularine improved punch wound closure in the ear pinna in mice. Both cytidine, a zebularine analogue, and uridine, a zebularine metabolite, did not promote ear pinna hole closure. The effect of zebularine on ear pinna healing...
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Interactions of antitumor triazoloacridinones with DNA
PublicationW niniejszej pracy zbadano oddziaływanie sześciu wybranych pochodnych triazoloakrydonu z DNA. Analiza wiskozymetryczna pokazała, że związki te interkalują do DNA. Jednocześnie dalsze badania wykazały, że proces interkalacji nie jest istotny aktywności cytotoksycznej tej grupy związków. Badania z wykorzystaniem fibroblastów dowiodły, że jedna z pochodnych triazoloakrydonu, związek C-1305, po uprzedniej metabolicznej aktywacji indukuje...
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Abstract DNA-type systems
PublicationAbstrakcyjny system typu DNA jest definiowany przez zestaw nieliniowych kinetycznych równań z wielomianowymi nieliniowościami, które przyjmują rozwiązania solitonowe związane z geometrią helis. Zestaw równań pozwala na dwie różne reprezentacje Laxa: forma von Neumanna i Darboux-kowariantna para Laxa. Wyjaśniamy dlaczego nie-Kolmogorowskie modele prawdopodobieństwa pojawiające się w kinetyce solitonowej są naturalnie związane z...
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An MTA-phosphorylase gene discovered in the metagenomic library derived from Antarctic top soil during screening for lipolytic active clones confers strong pink fluorescence in the presence of rhodamine B.
PublicationIn this work, we present the construction of a metagenomic library in Escherichiacoli using the pUC19 vector and environmental DNA directly isolated fromAntarctic topsoil and screened for lipolytic enzymes. Unexpectedly, the screeningon agar supplemented with olive oil and rhodamine B revealed one unusual pinkfluorescent clone (PINKuv) out of 85 000 clones. This clone harbored a plasmid,pPINKuv, which has an insert of 8317 bp that...
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Identification and molecular modeling of a novel lipase from an Antarctic soil metagenomic library.
PublicationIn this work, we present the construction of a metagenomic library in Escherichia coli using pUC19 vector and environmental DNA directly isolated from Antarctic topsoil and screened for lipolytic enzymes. The screening on agar supplemented with olive oil and rhodamine B revealed one clone with lipolytic activity (Lip1) out of 11,000 E. coli clones. This clone harbored a plasmid, pLip1, which has an insert of 4722 bp that has been...
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Effect of Ethanol Extracts of Propolis (EEPs) against Staphylococcal Biofilm—Microscopic Studies
PublicationStaphylococci growing in the form of biofilm exhibit high resistance to a plethora of antibiotics. The aim of the study was to assess the influence of ethanolic extract of propolis (EEPs) on S. epidermidis ATCC 35984 biofilm using fluorescent microscopy. Propidium iodide (PI) and SYTO 9 were used for differentiation of live and dead cells, and calcofluor white was used to stain the extracellular matrix, the self-produced extracellular...
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Conservation and diversity of MutS proteins
PublicationThe homologues of MutS, mismatch repair protein, exist in all prokaryotes, with the exception of Actinobacteria, Mollicutes and part of the Archaea. Multiple alignments of 316 MutS amino acid sequences from 169 species revealed conserved residues and sequence motifs distinguishing MutS homologues. All MutS homologues show high conservation within the ATPase domain. MutS1, the homologue responsible for DNA mismatch recognition,...
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The effect of sunscreen 4‐methylbenzylidene camphor in different and reproductive models, its bioaccumulation and molecular effects on ligand‐receptor interaction, and protein expression
Publication4-Methylbenzylidene camphor (4-MBC) is a photo-absorbing UV filter prevalently used in cosmetics, which can be absorbed into circulation and cause systemic effects. 4-MBC is continued to be released in the environment despite the growing knowledge about its bioaccumulation and endocrine disrupting effects. Previous reviews have mentioned UV-filter together but this review considers 4-MBC alone, due to its prevalence and concerning...
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IFI16 Impacts Metabolic Reprogramming during Human Cytomegalovirus Infection
PublicationCellular lipid metabolism plays a pivotal role in human cytomegalovirus (HCMV) infection, as increased lipogenesis in HCMV-infected cells favors the envelopment of newly synthesized viral particles. As all cells are equipped with restriction factors (RFs) able to exert a protective effect against invading pathogens, we asked whether a similar defense mechanism would also be in place to preserve the metabolic compartment from HCMV...
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Processing of 1-nitroacridine-induced DNA-DNA cross-links by topoisomerase I is associated with enhanced cellular survival: a possible role of topoisomerase I in the removal of DNA cross-links
PublicationW tej pracy wykazujemy, że modyfikacje plazmidu indukowane przez wzrastające dawki 1-nitroakrydyn (nitracrine i związek C-857) prowadzą do zahamowania aktywności katalitycznej topoizomerazy I. Hamowanie to związane jest z pojawieniem się pojedynczoniciowych uszkodzeń DNA i tworzeniem kowalencyjnych kompleksów topoizomeraza I-DNA. Nie obserwowano takich efektów dla topoizomerazy typu II.Dalsze badania wykazały, że jedna z 1-nitroakrydyn...
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Applying fuzzy logic to decisional DNA digital TV
PublicationWprowadzono wspomaganie oparte na logice rozmytej do modelu telewizji wykorzystujacej decyzyjny DNA.
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Bioaccumulation of Metals in Tissues of Marine Animals, Part I: the Role and Impact of Heavy Metals on Organisms
PublicationHeavy metals contribute to the anthropogenic contamination of marine ecosystems. Some of them are essential to the life processes of organisms; others are toxic, even at low concentrations. They penetrate organisms via food, respiratory pathways or the skin. The extent to which metals penetrate organisms is measured by the bioconcentration and bioaccumulation factors and also by their transport between organisms at different trophic...
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Naturally occurring mitochondrial DNA heteroplasmy in the MRL mouse
PublicationThe MRL/MpJ mouse is an inbred laboratory strain of Mus musculus, known to exhibit enhanced autoimmunity, increased wound healing, and increased regeneration properties. We report the full-length mitochondrial DNA (mtDNA) sequence of the MRL mouse (Accession # EU450583), and characterize the discovery of two naturally occurring heteroplasmic sites. The first is a T3900C substitution in the TpsiCloop of the tRNA methionine gene...
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Fuzyjne polimerazy DNA – otrzymywanie, charakterystyka i zastosowanie
PublicationObecnie reakcje PCR (ang. Polymerase Chain Reaction) wykazują bardzo szerokie zastosowanie w diagnostyce medycznej, biologii molekularnej czy inżynierii genetycznej. Efektywność tych reakcji rozumiana jako wydajność i wierność przeprowadzonej amplifikacji jest nieodłącznie związana ze stosowaną polimerazą DNA i warunkami prowadzenia reakcji PCR. Aby sprostać wymaganiom stawianym przez nowoczesne metody diagnostyczne oraz współczesną...
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Cold-adapted bacterial lipolytic enzymes and their applications
PublicationLipolytic 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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DNA modification by 1-nitroacridine derivatives. Comparison of 32P-Postlabeling and restriction enzyme analysis method for the detection of DNA adducts formed by 1-nitroacridines C-1748 and C-857
Publication4-podstawione 1-nitroakrydyny reprezentują nową grupę pochodnych akrydyny, zsyntetyzowanych na Politechnice Gdańskiej. Metoda 32P-postlabellingu wykazała, że pochodne C-1748 i C-857 tworzą od 4-6 adduktów DNA w systemie komórkowym i bezkomórkowym o innym rozkładzie plam chromatograficznych, jednakże niezależnym od zastosowanego systemu aktywacyjnego. Metoda enzymatycznej analizy restrykcyjnej pozwoliła natomiast śledzić kinetykę...
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A novel cold-active beta-D-galactosidase from the Paracoccus sp. 32d - gene cloning, purification and characterization
PublicationBeta-D-galactosidase (EC 3.2.1.23) catalyze the hydrolysis of terminal non-reducing beta-D-galactose residues in beta-D-galactosides. Cold-active beta-D-galactosidases have recently become a focus of attention of researchers and dairy product manufactures owing to theirs ability to: (I) eliminate of lactose from refrigerated milk for people afflicted with lactose intolerance, (II) convert lactose to glucose and galactose which...
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Interstrand crosslinking of DNA by C-1311 (Symadex) and other imidazoacridinones
PublicationImidazoakrydony, w tym pochodna C-1311 (Symadex) znajdująca się w pierwszej fazie badań klinicznych prowadzonych w USA, należą do grupy bardzo aktywnych związków przeciwnowotworowych zsyntetyzowanych w Katedrze Technologii Leków i Biochemii PG. Za pomocą zmodyfikowanej metody Parsons'a wykazaliśmy, że C-1311 indukuje, w sposób zależny od stężenia, międzyłańcuchowe wiązania sieciujące w komórkach różnych nowotworów ludzkiego pochodzenia...
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Electroanalytical and spectroscopic procedures for examination of interactions between double stranded DNA and intercalating drugs
PublicationPrzedstawiono metodę elektrochemicznego opisu oddziaływań dsDNA ze związkiem biologicznie czynnym w warunkach, w których zarówno badany związek jak i cząsteczka DNA są w roztworze elektrolitu. Opracowano warunki pomiaru, gdzie ani związek, ani DNA nie adsorbuje na elektrodzie. Wykazano, podobnie jak podczas badania innymi metodami, że oddziaływania są dwojakiego rodzaju: elektrostatyczne i poporzez interkalację.
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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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Interaction of antitumor triazoloacridone C-1305 and its analogs with telomeric DNA
PublicationBadaliśmy oddziaływanie pochodnej triazoloakrydonu C-1305 i jego bliskich analogów strukturalnych z oligonukleotydami zawierającycmi sekwencje telomerowe TTAGGG oraz oligonukleotydami sfałdowanymi w strukturę G-kwadrupleksu. Wykazaliśmy, że spośród badanych związków pochodna C-1305 wykazała największą specyficzność w stosunku do telomerowego DNA. Związek ten stabilizował także strukturę G-kwadrupleksu, podczas gdy jego analogi...
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Extremophile Deinococcus geothermalis as potential source of proteolytic enzymes
PublicationAmong many extremozymes, thermophilic enzymes have attracted most attention during the past four decades. Such enzymes are of considerable industrial and biotechnological interest due to the fact that the enzymes are better suited for harsh industrial processes. There are many advantages of conducting industrial processes at high temperature, such as the increased solubility of many polymeric substrates, resulting in decreased...
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Cellular senescence induced by DNA damage and its role in carcinogenesis and chemotherapy
PublicationUszkodzenia DNA indukują w komórce różne procesy, w tym starzenie komórkowe, co może uruchamiać proces karcinogenezy jak i być związane z odpowiedzią na leki przecinowotworowe. Biologiczna rola starzenia komórkowego nie jest jednak jasna. Zasugerowano, że proces starzenia komórkowego może hamować proces karcinogenezy poprzez indukcję nieodwracalnego zahamowania wzrostu przez czynniki genotoksyczne. Indukcja starzenia komórkowego...
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Detoxification of the tricyclic antidepressant opipramol and its analog – IS-noh by UGT enzymes before and after activation by phase I enzymes in rat liver microsomes
PublicationThe present studies were carried out to evaluate the simultaneous one-pot metabolism of opipramol (IS-opi) and analog (IS-noh) by phase I and phase II enzymes present in rat liver microsomes (RLM) as an alternative to separate testing with recombinant enzymes. This approach allows for more time-saving and cost-effective screening of the metabolism of newly discovered drugs. We also considered that the lack of results for phase...
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Epigenetic Basis of Regeneration: Analysis of Genomic DNA Methylation Profiles in the MRL/MpJ Mouse
PublicationEpigenetic 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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Novel thermostable single-stranded DNA-binding protein (SSB) from Deinococcus geothermalis
PublicationW celu zbadania właściwości biochemicznych białka wiążącego się do jednoniciowego DNA (SSB) pochodzącego z Deinococcus geothermalis (DgeSSB), sklonowano gen ssb otrzymany w reakcji PCR, a następnie uzyskano wydajną biosyntezę białka DgeSSB. Gen składa się z 900 par zasad, kodujących białko złożone z 300 reszt aminokwasowych, o wyliczonej masie molekularnej równej 32.45 kDa. Sekwencja aminokwasowa wykazuje 43, 44% i 75% identyczności...
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GRA1 protein vaccine confers better immune response compared to codon-optimized GRA1 DNA vaccine
PublicationW publikacji oszacowano immunogenność i właściwości protekcyjne szczepionki anty-Toxoplasma gondii opartej na białku rekombinantowym GRA1 oraz dwóch rodzajach szczepionek DNA (plazmidu zawierającego gen kodujący białko GRA1 dzikiego typu oraz modyfikowanego genu kodującego białko GRA1). Uzyskane wyniki sugerują lepsze właściowści immunoprotekcyjne rekombinantowego białka antygenowego niż szczepionek DNA.
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Geologia dna morskiego 2021/2022
e-Learning CoursesKurs przeznaczony jest dla studentów I semestru studiów II stopnia kierunku Inżynieria Morska i Brzegowa na WOiO i WILIŚ. Tematyka kursu obejmuje zagadnienia geologii morza, procesów formujących dno morskie i oceaniczne a także strefę brzegową oraz skał powstających w środowisku morskim.
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Gates of Enzymes
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Mechanical properties of mosquito nets in the context of hernia repair
PublicationThe paper deals with issue of applying mosquito nets as implants in hernia repair, which have already been used in resource-poor developing countries. Uniaxial tensile tests have been conducted on polyester mosquito meshes in two orthogonal directions. Non-linear elastic constitutive laws parameters have been identified to be applied in dense net material models. Mechanical performance of tested mosquito nets has been compared...
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Mathematical modelling of implant in an operated hernia for estimation of the repair persistence
PublicationThis paper presents mathematical modelling of an implanted surgical mesh used in the repair process of the abdominal hernia. The synthetic implant is simulated by a membrane structure. The author provides a material modelling of the implant based on the dense net model appropriate for technical fabrics. The accuracy of the proposed solution is evaluated by comparing the simulations of the dynamic behaviour of the system with the...
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Inter Applied Chemistry Programme 6 A practical approach to novel industrial enzymes Dr Natasha Bozic
e-Learning CoursesInter Applied Chemistry Programme 6 - A practical approach to novel industrial enzymes Dr Natasha Bozic The purpose of the course is to introduce students to the technologies of industrial enzymes discovery and manufacturing and to provide in depth insight about the advantages of using enzyme preparations in industry. Lectures will cover the common workflows of industrial enzymology including following topics: general enzyme...
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EXPERIMENTS AND SIMULATIONS FOR LAPAROSCOPICVENTRAL HERNIA REPAIR IMPROVEMENT
PublicationThe authors conducted a wide experimental and numerical study in order to determine effective approaches in the laparoscopic VH repair. At first the study covered identification of mechanical properties of the abdominal fascia, implants, tacks and transabdominalsutures. Basis on these data, the one and two-dimensional mathematical models were formulated and a number of numerical static and dynamic analyses were carried out. Then...
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Generation and Characterization of a DNA-GCN4 Oligonucleotide-Peptide Conjugate: The Impact DNA/Protein Interactions on the Sensitization of DNA
PublicationRadiotherapy, the most common therapy for the treatment of solid tumors, exerts its effects by inducing DNA damage. To fully understand the extent and nature of this damage, DNA models that mimic the in vivo situation should be utilized. In a cellular context, genomic DNA constantly interacts with proteins and these interactions could influence both the primary radical processes (triggered by ionizing radiation) and secondary reactions,...
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Interstrand crosslinking of DNA in tumor and fibroblast cells by C-1311 (Symadex) and other imidazoacridinones
PublicationImidazoakrydony, w tym pochodna C-1311 znajdująca się w pierwszej fazie badań klinicznych prowadzonych przez firmę Xanthus Life Sciences, należą do grupy bardzo aktywnych związków przeciwnowotworowych zsyntetyzowanych w Katedrze Technologii Leków i Biochemii PG. Za pomocą zmodyfikowanej metody Parsons'a wykazaliśmy, że C-1311 indukuje w sposób zależny od stężenia międzyłańcuchowe wiązania sieciujące w komórkach różnych nowotworów...
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Orthotropic membrane as a mechanical model of surgical implant in abdominal hernia repair
PublicationEven though the incisional hernia repair surgery is a well known procedure, mechanical properties of the tissue-implant system are unknown so the implantation of the repairing mesh is quite intuitive and, recurrences of the illness still take place. The main objective of the study is to define an operated hernia model that can be used for surgery planning and the assessment of the repair persistence. The load applied to the structure...
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IMPLANT USED IN HERNIA REPAIR UNDER PHYSIOLOGICAL MOVEMENTS OF HUMAN ABDOMEN
PublicationThe study deals with the mechanical behaviour of implant used in repair of abdominal hernia. The influence of orientation of the orthotropic implants in different areas of abdominal wall is studied.
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Assessment of usefulness of different tacks in laparoscopic ventral hernia repair
PublicationLaparoscopic ventral hernia repair is becoming a popular technique with good results and fast postoperative recovery. The mesh is placed directly under the peritoneum and anchored with transabdominal sutures and tacks. However, the ideal size of the mesh covering the hernia orifice is know, nor the ideal type or amount of tacks has to be described.