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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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Optimized 5-hour multiplex PCR test for the detection of tinea unguium: performance in a routine PCR laboratory
PublicationWe recently reported the development of a 5-hour multiplex PCR test for the detection of tinea unguium and the optimization of this test by the inclusion of an inhibition control. Here we report the performance of this procedure as used in a routine clinical laboratory as compared to conventional microscopy and culture-based techniques performed in a mycology reference laboratory. We found in processing 109 samples that 22 (20.2%)...
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In vitro and in silico assessment of anti-inflammatory activity of cocoa powders
PublicationPlants are considered the major sources of biologically active compounds, which provide unlimited opportunities for their use either as medical treatments or as novel drug formulations. Cocoa powder is frequently used in nutrition and is known to have many benefits thanks to its wide range of biological activities. The presented study was focused on th evaluation of the anti-inflammatory potential of extracts obtained from cocoa...
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GREENHIB Study of the impact of green corrosion inhibitors for aluminum alloys
ProjectsProject realized in Department of Electrochemistry, Corrosion and Materials Engineering according to 0675/IP2/2016/74 agreement from 2016-10-17
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The influence of G2 arrest abrogation on the long-term cytotoxicity of different genotoxic lesions
PublicationJak się uważa, zdolność komórek nowotworowych z uszkodzeniami DNA do zatrzymywania progresji w cyklu komórkowym w fazie G2 wydłuża czas na reperację tych uszkodzeń i zwiększa zdolność do przeżycia tych komórek. W pracy badaliśmy wpływ skrócenia bloku w fazie G2 w komórkach dwóch typów białaczek, poddanych działaniu związków przeciwnowotworowych indukujących różne typy uszkodzeń DNA, na efekt cytotoksyczny tych związków. Zwiększenie...
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Prediction of the structures of proteins with the UNRES force field, including dynamic formation and breaking of disulfide bonds
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Contactless Hearing Aid for Infants Employing Signal Processing Algorithms. [Bezkontaktowy aparat słuchowy dla niemowląt wykorzystujący algorytmy przetwarzania sygnału]
PublicationZaprojektowany bezkontaktowy aparat słuchowy umiejscawiany jest w łóżeczku niemowlęcia. Aparat składający się z matrycy 4 mikrofonów oraz prototypowej karty z procesorem DSP pracuje w polu swobodnym. Przetworzony sygnał mowy emitowany jest z wykorzystaniem miniaturowych głośników. Opracowane algorytmy pozwalają na elminację akustycznych sprzężeń zwrotnych, które mogą wystepować ze względu na niewielką odległość mikrofonów od głośników...
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Enzymatic activities behind degradation of glucosinolates
PublicationMyrosinase (thioglucosidase, EC 3.2.1.147) is the enzyme responsible for the hydrolysis of glucosinolates (GLs). In plant tissue, myrosinase and GLs are sequestered in separate cellular compartments. As a result of cell disruption, e.g., after pathogen attack or on chopping or grinding during food preparation, the myrosinase comes into contact with GLs and catalyzes the hydrolysis of thioglucosidic bond in GL structure. Consequently,...
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Isolation and some properties of collagen from the backbone of Baltic cod(Gadus morhua)
PublicationOssein from Baltic cod backbone was obtained after extraction of non-collagenous protein with 0.1 M NaOH solution and demineralization with 1.0 M HCl solution. The extractions were performed at 4 C for24, 48 and 72 h using a solid/solution ratio from 1:4 to 1:8 (w/v). After 48 h of extraction in 0.5 M acetic acid only about 25% of collagen was dissolved. After 48 h of extraction at optimal concentration of pepsin(4 mg/g ossein)...
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Approaches to Static Digestion Models
PublicationIt is not possible to look in detail at the wide range of static digestion methods that have been used to date. However, this section looks at some of the general approaches that have been used to look at the digestion of various nutrients and bioactives. I have focussed on the two main nutrients that undergo digestion in the upper GI tract, namely protein and lipid. In the case of protein, the research has largely been driven...
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Inhibition of amyloid fibril formation of hen egg white lysozyme by trimethylamine N-oxide at low pH
PublicationIn 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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Cancer-selective, single agent chemoradiosensitising gold nanoparticles
PublicationTwo nanometre gold nanoparticles (AuNPs), bearing sugar moieties and/or thiol-polyethylene glycol-amine (PEG-amine), were synthesised and evaluated for their in vitro toxicity and ability to radiosensitise cells with 220 kV and 6 MV X-rays, using four cell lines representing normal and cancerous skin and breast tissues. Acute 3 h exposure of cells to AuNPs, bearing PEG-amine only or a 50:50 ratio of alpha-galactose derivative and...
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Synthesis of Combretastatin A-4 Analogs and their Biological Activities
PublicationCombretastatin A-4 (CA-4) is a natural product, which consists of two phenyl rings, linked by an ethylene bridge. CA-4, inhibitor of polymerization of tubulin to microtubules, possesses a strong antitumor and anti-vascular properties both in vitro and in vivo. Previous studies showed that disodium phosphate salt of CA-4, a water-soluble prodrug is well tolerated at therapeutically useful doses. However, it should be noted that...
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Demonstrator technologii C-IED
PublicationArtykuł przedstawia budowę i działanie opracowanego na Politechnice Gdańskiej demonstratora technologicznego systemu AEGIS, przeznaczonego do przeciwdziałania atakom z wykorzystaniem prowizorycznych urządzeń wybuchowych detonowanych drogą radiową (RCIED — Radio Controlled Improvised Explosive Devices). Przedstawiono zaimplementowaną oryginalną metodę generowania sygnału zagłuszającego o dowolnie ukształtowanym widmie częstotliwościowym,...
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Benzonaza(R): możliwości praktycznych zastosowań.
PublicationBenzonaza znajduje zastosowanie w efektywnym obniżaniu lepkości i usuwaniu kwasów nukleinowych z roztworów białkowych. Zbadano w praktyce jej możliwości aplikacyjne w oczyszczaniu białek rekombinantowych wiążących jednoniciowe DNA (ang. SSB - single stranded DNA binding protein).
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Singlet oxygen-dominated peroxymonosulfate activation by layered crednerite for organic pollutants degradation in high salinity wastewater
PublicationAdvanced oxidation processes have been widely studied for organic pollutants treatment in water, but the degradation performance of radical-dominated pathway was severely inhibited by the side reactions between the anions and radicals, especially in high salinity conditions. Here, a singlet oxygen (1O2)-dominated non-radical process was developed for organic pollutants degradation in high salinity wastewater, with layered crednerite...
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The Effect of Conjugation with Octaarginine, a Cell-Penetrating Peptide on Antifungal Activity of Imidazoacridinone Derivative
PublicationAcridine cell-penetrating peptide conjugates are an extremely important family of compounds in antitumor chemotherapy. These conjugates are not so widely analysed in antimicrobial therapy, although bioactive peptides could be used as nanocarriers to smuggle antimicrobial compounds. An octaarginine conjugate of an imidazoacridinone derivative (Compound 1-R8) synthetized by us exhibited high antifungal activity against reference...
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Evaluation of antimicrobial activities of powdered cuttlebone against Klebsiella oxytoca, Staphylococcus aureus, and Aspergillus flavus
PublicationBackground: The presence of medicines in the environment is considered as a serious threat to the human health. The entrance of these substances into the water sources causes soil pollution, which eventually leads to the environmental pollution and it creates some problems for the public health. Also, increasing antibiotic resistant bacteria has attracted the attention of researchers to the use of natural resources such as marine...
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Investigation of the Antifungal Activity and Mode of Action of Thymus vulgaris, Citrus limonum, Pelargonium graveolens, Cinnamomum cassia, Ocimum basilicum, and Eugenia caryophyllus Essential Oils
PublicationThe antimicrobial activity of plant oils and extracts has been recognized for many years. In this study the activity of Thymus vulgaris, Citrus limonum, Pelargonium graveolens, Cinnamomum cassia, Ocimumbasilicum, and Eugenia caryophyllus essential oils (EOs) distributed by Pollena Aroma (Nowy Dwór Mazowiecki, Poland) was investigated against a group of 183 clinical isolates of C. albicans and 76 isolates of C. glabrata. All of...
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Regulacja topologii DNA przez topoizomerazę typu II, czyli jak struktura DNA wpływa na funkcjonowanie genów
PublicationW wielu ważnych procesach jakie zachodzą w komórce zmienia się topologia DNA i struktura chromatyny. Enzym, topoizomeraza typu II, jest jednym z ważnych czynników, który reguluje zmiany topologiczne i strukturalne w jądrze, przez co ma bezpośredni lub pośredni wpływ na takie procesy jak uporządkowanie struktury chromatyny poprzez wiązanie pętli chromatyny do szkieletu jądrowego, transkrypcję i replikację DNA, reperację uszkodzeń...
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A Highly Selective Biosensor Based on Peptide Directly Derived from the HarmOBP7 Aldehyde Binding Site
PublicationThis paper presents the results of research on determining the optimal length of a peptide chain to eectively bind octanal molecules. Peptides that map the aldehyde binding site in HarmOBP7 were immobilized on piezoelectric transducers. Based on computational studies, four Odorant Binding Protein-derived Peptides (OBPPs) with dierent sequences were selected. Molecular modelling results of ligand docking with selected peptides were...
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Novel single-stranded DNA-binding proteins from extreme psychrophilic bacterium Psychromonas ingrahamii 37
PublicationWe report the identification and characterization of the single-stranded DNA binding protein (SSB) from extreme psychrophilic bacterium Psychromonas ingrahamii 37 (PinSSB) that grows expotentially at -12°C and may well grow at even lower temperatures. PinSSB is one of the largest known bacterial SSB protein consisting 222 amino acid residues with a calculated molecular mass of 25.1 kDa. The analysis by gel filtration chromatography...
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Synthesis of 5-Substituted 1H-Tetrazoles from Nitriles in the Presence of Heterogeneous Catalyst
PublicationTetrazoles are five-membered heterocyclic compounds containing in their rings four nitrogen atoms. They have wide applications as corrosion inhibitors, analytical reagents, high-energy materials and gas generating compositions. Tetrazoles also play important role in coordination chemistry as ligands and in medicinal chemistry as metabolically stable surrogates for carboxylic acids. In recent years, investigation of new method of...
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Purification and biochemical characteristic of a cold-active recombinant esterase from Pseudoalteromonas sp. 643A under denaturing conditions.
PublicationIn this paper production of a cold-active esterase EstA from the Antarctic bacterium Pseudoalteromonas sp. 643A in E. coli expression system was described. The purification and biochemical characteristic of EstA were performed in the presence of urea and then compared with results obtained for the esterase with no addition of urea and isolated from the native source. In both cases the cold-active enzyme displayed similar properties....
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Utilizing Genome-Wide mRNA Profiling to Identify the Cytotoxic Chemotherapeutic Mechanism of Triazoloacridone C-1305 as Direct Microtubule Stabilization
PublicationRational drug design and in vitro pharmacology profiling constitute the gold standard in drug development pipelines. Problems arise, however, because this process is often dicult due to limited information regarding the complete identification of a molecule’s biological activities. The increasing aordability of genome-wide next-generation technologies now provides an excellent opportunity to understand a compound’s diverse eects...
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Mechanism of reaction of RNA-dependent RNA polymerase from SARS-CoV-2
PublicationWe combine molecular dynamics, statistical mechanics, and hybrid quantum mechanics/molecular mechanics simulations to describe mechanistically the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA-dependent RNA polymerase (RdRp). Our study analyzes the binding mode of both natural triphosphate substrates as well as remdesivir triphosphate (the active form of drug), which is bound preferentially over ATP by RdRp while...
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Sporothrix schenckii: purification and partial biochemical characterisation of glucosamine-6-phosphate synthase, a potential antifungal target
PublicationW artykule opisano izolację i charakterystykę właściwości natywnego enzymu, syntazy GlcN-6-P z komórek dimorficznego grzyba patogennego Sporothrix schenckii. Masę cząsteczkową enzymu natywnego określono na 350 kDa, a enzymu zdenaturowanego na około 79 kDa, co sugeruje homotetrameryczną strukturę białka. Wartość punktu izoelektrycznego wyniosła 6,26; wyznaczono także parametry kinetyczne katalizowanej reakcji. Aktywność enzymu była...
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KONSTRUKCJA AUKSOTROFICZNEGO WZGLĘDEM L-LIZYNY SZCZEPU CANDIDA ALBICANS
PublicationEnzymy szlaku kwasu L-α-aminoadypinowego biosyntezy L-lizyny z Candida albicans są potencjalnymi celami molekularnymi w terapii przeciwgrzybowej. Niektóre grupy badawcze wykazały, że auksotrofia wobec L-lizyny u pewnych gatunków grzybów powoduje zmniejszenie ich zjadliwości i może mieć wpływ na wzrost in vivo [Liebmann i in. 2004]. Oznacza to, że można się spodziewać zahamowania wzrostu komórek grzybowych w wyniku działania selektywnych...
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Functional domains of Candida albicans glucosamine-6-phosphate synthase.
PublicationSkonstruowano oddzielnie dwie domeny syntazy GlcN-6-P z Candida albicans jako białka fuzyjne, tj. domenę glutaminazową oraz domenę izomerazową, zawierające sekwencje poliHis, odpowiednio na C- i N-końcu. Zoptymalizowano warunki nadekspresji domen w komórkach Escherichia coli. Oba białka oczyszczono do homogenności przy zastosowaniu chromatografii metalopowinowactwa oraz zbadano ich podstawowe właściwości. Wyznaczono parametry kinetyczne...
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Evaluation of the yield, chemical composition and biological properties of essential oil from bioreactor-grown cultures of Salvia apiana microshoots
PublicationMicroshoot cultures of the North American endemic Salvia apiana were established for the first time and evaluated for essential oil production. Stationary cultures, grown on Schenk-Hildebrandt (SH) medium, supplemented with 0.22 mg/L thidiazuron (TDZ), 2.0 mg/L 6-benzylaminopurine and 3.0% (w/v) sucrose, accumulated 1.27% (v/m dry weight) essential oil, consisting mostly of 1,8-cineole, β-pinene, α-pinene, β-myrcene and camphor....
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MODELING THE pH EFFECTS ON NITROGEN REMOVAL IN THE ANAMMOX-ENRICHED GRANULAR SLUDGE
PublicationThe aim of the study was to determine the pH effects on nitrogen removal in the anammox-enriched 10 granular sludge. The experimental data were extracted from a 4L completely-mixed batch reactor with the 11 granular sludge at different initial pH values (6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5) and constant temperature 12 T=30℃. Simulations were run in GPS-X 6.4 using a comprehensive mechanistic model Mantis2. Two 13 kinetic parameters,...
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Modeling the pH effects on nitrogen removal in the anammox-enriched granular sludge
PublicationThe aim of the study was to determine the pH effects on nitrogen removal in the anammox-enriched granular sludge. The experimental data were extracted from a 4L completely-mixed batch reactor with the granular sludge at different initial pH values (6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5) and constant temperature T=30℃. Simulations were run in GPS-X 6.4 using a comprehensive mechanistic model Mantis2. Two kinetic parameters, including...
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Comparative Analysis of Phytochemical Profiles and Selected Biological Activities of Various Morphological Parts of Ligustrum vulgare
PublicationLigustrum vulgare (LV), widely cultivated in Europe and often used in hedges, has been histori-cally recognized in folk medicine for its potential health benefits. This study focused on exploring the untargeted identification of secondary metabolites in ethanol extracts (70% v/v) from differ-ent morphological parts (young shoots, leaves, flowers and fruits) of LV at various stages of plant development, using ultra-high-performance...
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High-performance activation of ozone by sonocavitation for BTEX degradation in water
PublicationThis work presents a novel advanced oxidation process (AOP) for degradation of emerging organic pollutants – benzene, toluene, ethylbenzene and xylenes (BTEXs) in water. A comparative study was performed for sonocavitation assisted ozonation under 40–120 kHz and 80–200 kHz dual frequency ultrasounds (DFUS). Based on the obtained results, the combination of 40–120 kHz i.e., low-frequency US (LFDUS) with O3 exhibited excellent oxidation...
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MnCo2O4 deposited by spray pyrolysis as a protective layer for stainless steel interconects
PublicationStainless steel interconnects working in Solid Oxide Fuel Cells stacks are exposed to high temperature resulting in their corrosion. Protective layers for the hydrogen and oxygen sides are necessary to protect the interconnect material, prolongate the stack lifetime and maintain the output power. In this paper MnCo2O4 protective layer for the oxygen side of the interconnect is deposited by spray pyrolysis and is examined.
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It applications for the remote testing of hearing.
PublicationTelemedycyna odgrywa coraz wiekszą rolę w diagnostyce i leczeniu osób z ubytkami słuchu. Jest to związane m.in. ze specyfiką badań audiometrycznych. Postęp technologiczny w dziedzinie aparatów słuchowych i implantów ślimakowych wymusza nowe metody diagnozy w audiologii, jak również w praktyce otolaryngologicznej. Serwis ''Telezdrowie'', w którym zaimplementowano liczne testy przesiewowe jest przykładem prowadzenia diagnostyki w...
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Hearing aid operating in acoustical free field
PublicationAparatowanie bardzo małych dzieci (od 5 miesiąca życia) za pomocą standardowych protez słuchu natrafia na wiele trudności natury praktycznej. Dotyczy to procesu dopasowania aparatu słuchowego, czyli doboru jego ustawień stosownie do aktualnych charakterystyk ubytku słuchu dzieci. Tymczasem wczesne aparatowanie jest zagadnieniem o ogromnym zanczeniu dla rozwoju słuchu, mowy i ogólnej inteligencji dziecka. Referat prezentuje uzyskane...
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Tereny polarne - interesującym obszarem do badań Cz. II - Analityka próbek abiotycznych w Arktyce
PublicationW pracy ujęto problematykę zanieczyszczeń obszarów polarnych. Zwrócono szczególną uwagę na trwałe związki organiczne obecne w rejonie Arktyki w elementach przyrody nieożywionej. TZO pochodzenia antropogenicznego mogą pojawiać się na terenach polarnych za sprawą transportu na duże odległości z obszarów Euroazji i Ameryki Północnej. Badania poziomów zanieczyszczeń obecnych w próbkach abiotycznych na obszarze Arktyki stanowią fundament...
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Kod fontannowy z przyrastającą liczbą symboli źródłowych
PublicationKody fontannowe zabezpieczające transmisję przed wymazaniami wyróżnia brak określonej z góry długości i losowy sposób generacji kolejnych pakietów kodowych. W pracy pokazano jak właściwości tych kodów zależą od rozkładu macierzy generującej dla dwóch wariantów dostarczania pakietów do kodera. Szczególnie interesujący jest przypadek, gdy pakiety źródłowe trafiają do kodera stopniowo podczas transmisji. Uzyskane wyniki wskazują na...
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Photoinactivation of Staphylococcus aureus using protoporphyrin IX: the role of haem-regulated transporter HrtA
PublicationLight- and photosensitiser-based antimicrobial photodynamic therapy is a very promising approach to the control of microbial infections. How the phenotypic features of a microorganism affect its response to photosensitiser-based photokilling represents an area of substantial research interest. To understand the mechanisms governing the phenomenon of a strain-dependent response to photodynamic inactivation (PDI), we analysed the...
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Genetic and pharmacologic proteasome augmentation ameliorates Alzheimer’s-like pathology in mouse and fly APP overexpression models
PublicationThe proteasome has key roles in neuronal proteostasis, including the removal of misfolded and oxidized proteins, presynaptic protein turnover, and synaptic efficacy and plasticity. Proteasome dysfunction is a prominent feature of Alzheimer’s disease (AD). We show that prevention of proteasome dysfunction by genetic manipulation delays mortality, cell death, and cognitive deficits in fly and cell culture AD models. We developed...
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A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter sp. 32c - gene cloning, overexpression, purification and properties
PublicationThe development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological...
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‘Acridines’ as New Horizons in Antifungal Treatment
PublicationFrequent fungal infections in immunocompromised patients and mortality due to invasive mycosis are important clinical problems. Opportunistic pathogenic Candida species remain one of the leading causes of systemic mycosis worldwide. The repertoire of antifungal chemotherapeutic agents is very limited. Although new antifungal drugs such as lanosterol 14α-demethylase and β-glucan synthase inhibitors have been introduced into clinical...
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Solid and liquid fractionation of sugarcane and Agave bagasse during ozonolysis and enzymatic hydrolysis: Impact on biohydrogen and biogas production
PublicationSustainable and cost-efficient lignocellulosic biomass-to-energy bioprocesses require assessing the main components released during biomass fractionation. This study assessed the mesophilic biochemical potential of biohydrogen (BHP) and methane production (BMP) from sugarcane bagasse (SCB) and cooked (CAB) and uncooked (UAB) Agave bagasse. A systematic comparative analysis was performed for the liquid and solid fractions derived...
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Structural, functional, and stability change predictions in human telomerase upon specific point mutations,
PublicationOverexpression of telomerase is one of the hallmarks of human cancer. Telomerase is important for maintaining the integrity of the ends of chromosomes, which are called telomeres. A growing number of human disease syndromes are associated with organ failure caused by mutations in telomerase (hTERT or hTR). Mutations in telomerase lead to telomere shortening by decreasing the stability of the telomerase complex, reducing its accumulation,...
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Triazoloacridone C-1305 abrogates the restriction checkpoint in cells lacking functional p53 and promotes their accumulation in the G2/M phase of the cell cycle
PublicationTriazoloakrydon C-1305, nowy inhibitor topoizomerazy II, wykazuje wysoką aktywość cytotoksyczną wobec komórek wielu typów nowotworów. Komórki mysie pozbawione genu PARP-1 wykazywały nadwrażliwość na działanie tego związku. W pracy testowaliśmy działanie związku wobec dwóch różnych komóek nowotworowych różniących się statusem p53: komórkami białaczki myelocytarnej HL-60 i raka sutka MCF-7. Wykazaliśmy, że inkubacja ze związkiem...
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Allosteric control of Candida albicans glucosamine-6-phosphate synthase.
PublicationPrzedstawiono wyniki badań spektrofluorymetrycznych syntazy GlcN-6-P z Candida albicans oraz enzymu zmutowanego, w którym reszty Trp388 i 690 zostały zamienione na drodze ukierunkowanej mutagenezy na reszty Phe. Metodą spektrofluorymetryczną badano wpływ ligandów: Fru-6-P (substrat), Gln (substrat), UDP-GlcNAc (inhibitor), Glc-6-P (aktywator), DON (analog glutaminy) w odpowiednio dobranych stężeniach. Obecność większości efektorów...
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Obróbka biologiczna Investigations of the biological pre-treatment of lignocellulosic biomass on dark fermentation inhibitory compounds generation
ProjectsProject realized in Department of Process Engineering and Chemical Technology according to DEC-2021/05/X/ST8/00919 agreement from 2021-11-17
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Krajobraz nomadyczny w Muittalus samid birra (1910) Johana Turiego
PublicationThe article introduces Muittalus samid birra (1910, An account of the Sami) by Johan Turi, the fi rstbook with a secular content written in the sami language by a sami person. The author, a wolf hunterand reindeer-breeder, describes a broad spectrum of his people’s daily life. Pictures of everydayactivities are supplemented with stories about the sami people’s knowledge of the world (traditions,myths,...
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Young shoots of red cabbage are a better source of selected nutrients and glucosinolates in comparison to the vegetable at full maturity
PublicationCruciferous vegetables are a valuable source of ingredients with health benefits. The most characteristic compounds of cruciferous vegetables with identified anticancer properties are glucosinolates. Young shoots and sprouts of red cabbage are becoming a popular fresh food rich in nutrients and bioactive compounds. The objective of this research was to determine, for the first time in a comprehensive approach, whether young shoots...