The effect of thyme and tea tree oils on morphology and metabolism of Candida albicans

Members of Candida species cause significant problems in medicine and in many industrial branches also. In order to prevent from Candida sp. development, essential oils are more and more frequently applied as natural, non-toxic, non-pollutive and biodegradable agents with a broad spectrum of antimicrobial activity. The aim of the research was to determine changes in morphology and metabolic properties of Candida albicans in the presence of thyme and tea tree oils. Changes of enzymatic activity of isolates were observed in the presence of both tested essential oils, and they were primarily associated with loss or decrease of activity of all enzymes detected for control. Furthermore, only for 3 out of 11 isolates additional activity of N-acetyl-β-glucosaminidase, α-mannosidase, α-fucosidase and trypsin was detected. Vivid changes in biochemical profiles were found after treatment with tea tree oil and they were related to loss of ability to assimilate d-xylose, d-sorbitol and d-trehalose. The main differences in morphology of isolates compared to the control strain concerned formation of pseudohyphae structures. Both examined essential oils caused changes in cell and colony morphology, as well as in the metabolism of Candida albicans. However, the extent of differences depends on the type and concentration of an essential oil. The most important finding is the broad spectrum of changes in yeast enzymatic profiles induced by thyme and tea tree oils. It can be supposed that these changes, together with loss of ability to assimilate saccharides could significantly impact Candida albicans pathogenicity

Probiotička aktivnost kvasca Saccharomyces cerevisiae var. boulardii u sprečavanju patogenih bakterija

Infectious diarrhoea is associated with a modification of the intestinal microflora and colonization of pathogenic bacteria. Tests were performed for seven probiotic yeast strains of Saccharomyces cerevisiae var. boulardii, designated for the prevention and treatment of diarrhoea. To check their possible effectiveness against diarrhoea of different etiologies, the activity against a variety of human pathogenic or opportunistic bacteria was investigated in vitro. In mixed cultures with S. cerevisiae var. boulardii, a statistically significant reduction was observed in the number of cells of Listeria monocytogenes, Pseudomonas aeruginosa and Staphylococcus aureus, by even 55.9 % in the case of L. monocytogenes compared with bacterial monocultures. The influence of yeasts was mostly associated with the shortening of the bacterial lag phase duration, more rapid achievement of the maximum growth rates, and a decrease by 4.4–57.1 % (L. monocytogenes, P. aeruginosa), or an increase by 1.4–70.6 % (Escherichia coli, Enterococcus faecalis, Salmonella Typhimurium) in the exponential growth rates. Another issue included in the research was the ability of S. cerevisiae var. boulardii to bind pathogenic bacteria to its cell surface. Yeasts have shown binding capacity of E. coli, S. Typhimurium and additionally of S. aureus, Campylobacter jejuni and E. faecalis. However, no adhesion of L. monocytogenes and P. aeruginosa to the yeast cell wall was noted. The probiotic activity of S. cerevisiae var. boulardii against human pathogens is related to a decrease in the number of viable and active cells of bacteria and the binding capacity of yeasts. These processes may limit bacterial invasiveness and prevent bacterial adherence and translocation in the human intestines.Promjene u crijevnoj mikroflori uzrokuju naseljavanje patogenih bakterija i infektivni proljev. Ispitano je sedam probiotičkih sojeva kvasca Saccharomyces cerevisiae var. boulardii radi prevencije i liječenja proljeva. Istražena je in vitro aktivnost tih sojeva u sprečavanju različitih patogenih i oportunističkih bakterija da bi se utvrdila njihova učinkovitost pri liječenju različitih uzroka proljeva. U miješanim kulturama kvasca S. cerevisiae var. boulardii, za razliku od monokultura, opaženo je statistički značajno smanjenje broja bakterija Listeria monocytogenes (i to čak za 55,9 %), Pseudomonas aeruginosa i Staphylococcus aureus. Prisutnost kvasaca prije svega skraćuje fazu mirovanja bakterija (lag faza), zatim ubrzava postizanje maksimuma rasta, te smanjuje stopu eksponencijalnog rasta za 4,4-57,1 % (L. monocytogenes, P. aeruginosa) ili ju povećava za 1,4-70,6 % (Escherichia coli, Enterococcus faecalis, Salmonella Typhimurium). Osim toga, ispitana je sposobnost vezivanja patogenih bakterija na površinu stanica kvasca S. cerevisiae var. boulardii. Kvasci su uspješno vezali bakterije E. coli, S. Typhimurium, S. aureus, Campylobacter jejuni i E. faecalis, ali ne i L. monocytogenes i P. aeruginosa. Probiotičkom aktivnošću kvasca S. cerevisiae var. boulardii smanjuje se broj živih i aktivnih stanica patogenih bakterija koje su se vezale na stanice kvasca. Ti procesi ograničavaju invazivnost bakterija i sprečavaju njihovo vezanje za stijenke crijeva

Pre-columbian archeological textiles: A source of pseudomonas aeruginosa with virulence attributes

Pseudomonas aeruginosa is an opportunistic pathogen associated with a broad spectrum of infections in humans. However, the pathogenicity of environmental P. aeruginosa strains, especially isolates from museums and conservation laboratories, is not widely recognized. In this study, the virulence attributes of P. aeruginosa isolated from pre-Columbian textiles were compared to those of a clinical strain. Both genetically identified environmental strains (KP842564 and KP842565) exhibited a high ability to form biofilms on abiotic surfaces and high hemolytic activity. In addition, strain KP842564 was a moderate pyocyanin producer and showed proteolytic properties toward bovine serum albumin, fibrinogen, mucin, and casein. In contrast to the clinical isolate, the environmental strainswere susceptible to all the tested antimicrobial agents. The strains also showed high bioadhesion and colonization capacity on archeological textile samples, in which wool fibers were the only source of nutrients, as confirmed by scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX) analysis. This study highlights the need to identify microorganisms which inhabit historic objects, in order to avoid exposure to occupational hazards. Although the strain KP842565 exhibited only some of the examined virulence-related features, given that the production of pyocyanin and hemolysins as well as the formation of biofilm are important virulence factors of P. aeruginosa, the results indicate that these strains may present a potential health risk for humans.Fil: Rajkowska, Katarzyna. Lodz University of Technology; PoloniaFil: Otlewska, Anna. Lodz University of Technology; PoloniaFil: Guiamet, Patricia Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Wrzosek, Henryk. Lodz University of Technology; PoloniaFil: Machnowski, Waldemar. Lodz University of Technology; Poloni

The Trends and Prospects of Winemaking in Poland

Viticulture and winery origins in Poland date to the tenth century, but their tradition has been reborn in the last ten years, resulting in a development of small vineyards producing excellent wines not only for the local market. Due to the cold climate, usually short summers with moderate and low temperatures, the grapes are characterized by lower sugar content and higher acidity compared to those grown in the south of Europe. According to the European Union regulations, Poland was classified as the coldest wine-growing region (A) and officially acknowledged as a wine-producing country. The grapevine cultivars adopted to the harsh climatic conditions give the Polish grape wines some unique sensory features. The most popular varieties of grapes for the production of red wine are Regent, Rondo, Pinot Noir, Maréchal Foch, Cabernet Cortis, Tryumf Alzacji, Cascade and Dornfelder. For white wine production, Solaris, Riesling, Seyval Blanc, Pinot Gris, Johanniter, Jutrzenka, Hibernal, Aurora, Bianka, Traminer, Jutrzenka and Siberia are mostly used in Poland. This chapter presents Polish grape winery with its specificity and prospects for the future. The traditional products of Polish fermentation industry, fruit wines and meads, are also mentioned

Abiotic Determinants of the Historical Buildings Biodeterioration in the Former Auschwitz II – Birkenau Concentration and Extermination Camp

The paper presents the results of a study conducted at the Auschwitz-Birkenau State Museum in Oświecim on the occurrence of biodeterioration. Visual assessment of the buildings revealed signs of deterioration of the buildings in the form of dampness, bulging and crumbling plaster, and wood fiber splitting. The external surfaces, and especially the concrete strips and ground immediately adjoining the buildings, were colonized by bryophytes, lichens, and algae. These organisms developed most intensively close to the ground on the northern sides of the buildings. Inside the buildings, molds and bacteria were not found to develop actively, while algae and wood-decaying fungi occurred locally. The factors conducive to biological corrosion in the studied buildings were excessive dampness of structural partitions close to the ground and a relative air humidity of above 70%, which was connected to ineffective moisture insulation. The influence of temperature was smaller, as it mostly affected the quantitative composition of the microorganisms and the qualitative composition of the algae. Also the impact of light was not very strong, but it was conducive to algae growth

Polyoxometalate‐Ionic Liquids (POM‐ILs) as anticorrosion and antibacterial coatings for natural stones

Resumen del póster presentado al RSC Chemical Nanoscience and Nanotechnology Early Careers Virtual Meeting, celebrado el 24 de marzo de 2021.Peer reviewe

Phenotypic and genotypic diversity of wine yeasts used for acidic musts

The aim of this study was to examine the physiological and genetic stability of the industrial wine yeasts Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum under acidic stress during fermentation. The yeasts were sub-cultured in aerobic or fermentative conditions in media with or without l-malic acid. Changes in the biochemical profiles, karyotypes, and mitochondrial DNA profiles were assessed after minimum 50 generations. All yeast segregates showed a tendency to increase the range of compounds used as sole carbon sources. The wild strains and their segregates were aneuploidal or diploidal. One of the four strains of S. cerevisiae did not reveal any changes in the electrophoretic profiles of chromosomal and mitochondrial DNA, irrespective of culture conditions. The extent of genomic changes in the other yeasts was strain-dependent. In the karyotypes of the segregates, the loss of up to 2 and the appearance up to 3 bands was noted. The changes in their mtDNA patterns were much broader, reaching 5 missing and 10 additional bands. The only exception was S. bayanus var. uvarum Y.00779, characterized by significantly greater genome plasticity only under fermentative stress. Changes in karyotypes and mtDNA profiles prove that fermentative stress is the main driving force of the adaptive evolution of the yeasts. l-malic acid does not influence the extent of genomic changes and the resistance of wine yeasts exhibiting increased demalication activity to acidic stress is rather related to their ability to decompose this acid. The phenotypic changes in segregates, which were found even in yeasts that did not reveal deviations in their DNA profiles, show that phenotypic characterization may be misleading in wine yeast identification. Because of yeast gross genomic diversity, karyotyping even though it does not seem to be a good discriminative tool, can be useful in determining the stability of wine yeasts. Restriction analysis of mitochondrial DNA appears to be a more sensitive method allowing for an early detection of genotypic changes in yeasts. Thus, if both of these methods are applied, it is possible to conduct the quick routine assessment of wine yeast stability in pure culture collections depositing industrial strains

Antifungal Activity of Polyoxometalate-Ionic Liquids on Historical Brick

Moulds inhabiting mineral-based materials may cause their biodeterioration, contributing to inestimable losses, especially in the case of cultural heritage objects and architectures. Fungi in mouldy buildings may also pose a threat to human health and constitute the main etiological factor in building related illnesses. In this context, research into novel compounds with antifungal activity is of high importance. The aim of this study was to evaluate the antifungal activity of polyoxometalate-ionic liquids (POM-ILs) and their use in the eradication of moulds from historical brick. In the disc diffusion assay, all the tested POM-ILs inhibited growth of a mixed culture of moulds including Engyodontium album, Cladosporium cladosporioides, Alternaria alternata and Aspergillus fumigatus. These were isolated from the surfaces of historical brick barracks at the Auschwitz II-Birkenau State Museum in Oświęcim, Poland. POM-IL coatings on historical brick samples, under model conditions, showed that two compounds demonstrated very high antifungal activity, completely limiting mould growth and development. The antifungal activity of the POM-ILs appeared to stem from their toxic effects on conidia, as evidenced by environmental scanning transmission electron microscopy observations. The results herein indicated that POM-ILs are promising disinfectant materials for use not only on historical objects, but probably also on other mineral-based materials

Modificirane molekule U1 RNA kot korektorji procesiranja RNA pri humanih genetskih boleznih

The experimental work of this thesis was performed at the International Centre for Genetic Engineering and Biotechnology (ICGEB) in the Human Molecular Genetics Group, under the scientific direction of Prof. Franco Pagani. The project was developed during the academic years 2014-2017. Modified U1 RNAs, also named Exon Specific U1s (ExSpeU1s) represent a novel class of small RNA-based molecules that correct exons splicing defects. To evaluate their therapeutic potential focused on Familial Dysautonomia (FD), a rare autosomal recessive disorder characterized by progressive degeneration of the sensory and autonomic nervous system. More than 99% of patients are homozygous for the T to C transition in position 6 of the IKBKAP intron 20 (c.2204+6T>C). This substitution modifies the exon 20 5’ splice site (5’ss) inducing exon skipping in a tissue-specific manner and reducing the total amount of IKAP protein. The molecular mechanisms underlying the IKBKAP mis-splicing are not completely clear and there are no effective treatments. In this thesis, I investigated the therapeutic potential of ExSpeU1s and the role of cis- and trans-acting factors that regulates IKBKAP splicing. Using a splicing functional assay, I identified ExSpeU1s that bind to intron 20 sequences and rescue the exon 20 skipping defect. Interestingly, their rescue activity was modulated by several splicing factors and requires a critical exonic splicing enhancer element. Transfection experiment showed the involvement of both enhancing (TIA1, PTBP1 and PTB4) and inhibitory (SRSF3, hnRNPA1, FOX and FUS) splicing factors in IKBKAP splicing. To better evaluate the ExSpeU1s therapeutic efficacy, I transduced FD patient’s fibroblasts with a lentiviral vector expressing the most active ExSpeU1. This resulted in a complete rescue of the exon skipping defect and improvement in IKAP protein expression. Most importantly, intraperitoneal delivery of ExSpeU1s by AAV9 into a transgenic mouse model, that recapitulates the tissue-specific mis-splicing seen in FD patients, corrected the aberrant splicing patterns in several tissues increasing the amount of the corresponding IKAP protein. All together, these results identify novel regulatory splicing factors involved in the IKBKAP exon 20 regulation and provide the proof of principle that ExSpeU1s delivered in vivo by AAV vectors represent a novel therapeutic strategy for FD.Eskperimentalno delo je bilo opravljeno na Mednarodnem centru za genetski inženiring in biotehnologijo (International Centre fo Genetic Enginnerring and Biotechnology, ICGEB, Trst, Italija) v skupini za Humano molekularno genetiko, pod mentorstvom prof. Franco Pagani. Projekt je bil izveden v akademskem obdobju 2014-2017. Družinska disavtonomija (and. Familial Dysautonomia, FD), oz. sindrom Riley-Day, je redko avtosomalno recesivno dedno obolenje, ki je karakterizirano s progresivnim odmiranjem senzoričnega in avtonomnega živčnega sistema. Večinoma je prisotno v populaciji Ashkenazi Judov. Več kot 99% bolnikov ima hogozigotno obliko tranzicije T v C na mestu 6 introna 20 v genu IKBKAP (c.2204+6T>C). Ta nukleotidna zamenjava spremeni 5\u27 spojitveno mesto (5\u27ss) eksona 20, kar povzroči tkivno-specifično izključitev eksona v prepisu gena in posledično zmanjšano količino proteina IKAP. Molekularni mehanizmi, ki povzročajo spremembe v spajanju eksonov niso popolnoma poznani. Za bolezen ne obstajajo učinkovita zdravljenja. V doktorskem delu sem raziskovala terapevtski potencial nove skupine malih RNA molekul, ki se imenujejo ekson-specifične U1 molekule (ExSpeU1) in vpliv cis- in trans-delujočih faktorjev, ki regulirajo IKBKAP spajanje eksonov. Z uporabo funkcionalne metode spajanja sem določila ExSpeU1 molekule, ki se vežejo na zaporedja introna 20 in tako popravijo izključitev eksona 20 iz prepisa gena. Ta aktivnost je regulirana z več faktorji spajanja ter zaporedjem elementa eksonskega ojačevalca spajanja. Transfekcija celičnih linij je pokazala, da so v proces spajanja IKBKAP vključeni tako ojačevalci spajanja (TIA1, PTBP1 in PTB4) ter prav tako inhibitorni faktorji (SRSF3, hnRNPA1, FOX in FUS). Z namenom boljše določitve terapevtskega potenciala ExSpeU1, sem transducirala fibroblaste, ki so bili pridobljeni iz bolnika z FD ter uporabo lentivirusnih vektorjev, ki so izražali najbolj učinkovito ExSpeU1. To je povzročilo popolno popravilo izključitve eksona in tako izboljšanje izražanja proteina IKAP. Intraperitonalni prenos ExSpeU1 z AAV9 vektorjem v transgenem mišjem modelu, ki predstavlja tkivno-specifično okvaro spajanja podobno kot pri bolnikih FD, je popravilo vzorce okvarjenega spajanja v večih tkivih in tako povečalo izražanje pripadajočega IKAP proteina. Rezultati so določili nove regulatorne faktorje spajanja, ki so udeleženi v regulaciji spajanja IKBKAP eksona 20 in tako predstavljajo potrditev načela domneve, da ExSpeU1 molekule prenesene in vivo z AAV vektorji predstavljajo ustrezno terapevstko strategijo za bolnike FD

Stabilnost kvasaca Saccharomyces sensu stricto i njihovih hibrida tijekom fermentacije

The objective of this paper is to investigate the technological usefulness of selected industrial wine yeasts Saccharomyces cerevisiae and Saccharomyces bayanus and their intra- and interspecific hybrids responsible for excessively acidic musts. The stability of yeast fermentation profiles in apple musts was assessed after 90–170 generations, following previous subculturing under aerobic or anaerobic conditions in media with or without L-malic acid. During this study, 35 apple wines produced by wild strains and their segregates were statistically evaluated according to 12 chemical parameters. Although the wines met the official standards for basic chemical parameters, their total acidity was too low. Both the yeasts and their segregates metabolized from 66.3 to 77.0 % of malic acid present in the must. The industrial wine yeasts and their hybrids exhibited marked polymorphism of fermentation profiles in apple must with elevated L-malic acid content. At the same time, the level of demalication activity made it possible to clearly differentiate segregates from the wild strains, which may suggest that malic acid is probably one of the principal factors in the adaptive evolution of yeasts. Our study proves that among industrial wine yeasts, there are both, strains expressing very high stability (Saccharomyces cerevisiae W-13) and labile ones (S. cerevisiae Syrena). The interspecific hybrids S. cerevisiae × S. bayanus showed low stability of technological features, while the intraspecific hybrid of S. cerevisiae preserved its fermentative capacity. The presented results indicate that fermentative stability assessment under environmental stress can help to select the yeast strains best suited for the fermentation of specific musts.Svrha je rada bila istražiti učinkovitost odabranih industrijskih sojeva kvasaca Saccharomyces cerevisiae i Saccharomyces bayanus u proizvodnji vina te njihovih hibrida, dobivenih križanjem unutar i između vrsta, koji mogu uzrokovati preveliku kiselost mošta. Procijenjena je stabilnost kvasaca, prethodno uzgojenih u aerobnim ili anaerobnim uvjetima na podlozi koja sadržava L-jabučnu kiselinu ili bez nje, pri fermentaciji jabučnog mošta tijekom 90-170 generacija. Statistički je ispitano 12 kemijskih značajki 35 uzoraka jabučnih vina proizvedenih pomoću divljih sojeva kvasaca i sojeva izoliranih nakon uzastopnih precjepljivanja. Iako su osnovna kemijska svojstva vina zadovoljavala službene kriterije kakvoće, ukupna je kiselost bila premala. Kvasci su metabolizirali 66,3-77 % jabučne kiseline iz mošta. Industrijski sojevi vinskih kvasaca i njihovi hibridi izrazito su polimorfni, pa su različito fermentirali jabučni mošt s povećanim udjelom L-jabučne kiseline. Istodobno se prema postotku razgrađene jabučne kiseline mogu razlikovati izolati nakon uzastopnih precjepljivanja od divljih sojeva, što znači da je jabučna kiselina jedan od glavnih čimbenika koji utječe na evoluciju kvasaca. U radu je dokazano da među industrijskim vinskim kvascima ima sojeva koji su jako stabilni (Saccharomyces cerevisiae W-13), a i onih koji su labilni (S. cerevisiae Syrena). Hibrid nastao križanjem vrsta S. cerevisiae i S. bayanus nedovoljno je stabilan tijekom fermentacije, a onaj nastao križanjem unutar vrste S. cerevisiae zadržao je fermentativna svojstva. Rezultati potvrđuju da se procjenom stabilnosti kvasaca u uvjetima stresa mogu odabrati sojevi najprikladniji za fermentaciju pojedinih moštova