Authors:Silke Dubbert, Birgit Klinkert, Michael Schimiczek, Trudy M. Wassenaar, and Rudolf von Bünau
Probiotic Escherichia coli strain Nissle 1917 (EcN) has a long history of safe use. However, the recently discovered presence of a pks locus in its genome presumably producing colibactin has questioned its safety, as colibactin has been implicated in genotoxicity. Here, we assess the genotoxic potential of EcN. Metabolic products were tested in vitro by the Ames test, a mutagenicity assay developed to detect point mutation-inducing activity. Live EcN were tested by an adapted Ames test. Neither the standard nor the adapted Ames test resulted in increased numbers of revertant colonies, indicating that EcN metabolites or viable cells lacked mutagenic activity. The in vivo Mammalian Alkaline Comet Assay (the gold standard for detecting DNA-strand breaks) was used to determine potentially induced DNA-strand breaks in cells of the gastro-intestinal tract of rats orally administered with viable EcN. Bacteria were given at 109–1011 colony forming units (CFU) per animal by oral gavage on 2 consecutive days and daily for a period of 28 days to 5 rats per group. No significant differences compared to negative controls were found. These results demonstrate that EcN does not induce DNA-strand breaks and does not have any detectable genotoxic potential in the test animals.
Authors:Jakob Knorr, Steffen Backert, and Nicole Tegtmeyer
The gastric pathogen Helicobacter pylori colonizes approximately half of the human world population. The bacterium injects the effector protein cytotoxin associated gene A (CagA) via a type-IV secretion system into host epithelial cells, where the protein becomes phosphorylated at specific EPIYA-motifs by cellular kinases. Inside the host cell, CagA can interact with over 25 different proteins in both phosphorylation-dependent and phosphorylation-independent manners, resulting in manipulation of host-cell signaling pathways. During the course of an H. pylori infection, certain host-cell proteins undergo tyrosine dephosphorylation in a CagA-dependent manner, including the actin-binding proteins cortactin and vinculin. A predominant response of intracellular CagA is the binding and activation of tyrosine phosphatase, the human Src-homology-region-2-domain-containing-phosphatase-2 (SHP2). Here, we considered the possibility that activated SHP2 might be responsible for the dephosphorylation of cortactin and vinculin. To investigate this, phosphatase inhibitor studies were performed. Additionally, a complete knockout mutant of SHP2 in AGS cells was created by CRISPR/Cas9 technology, and these cells were infected with H. pylori. However, neither the presence of an inhibitor nor the inactivation of SHP2 prevented the dephosphorylation of cortactin and vinculin upon CagA delivery. Tyrosine dephosphorylation of these proteins is therefore independent of SHP2 and instead must be caused by another, as yet unidentified, protein tyrosine phosphatase.
Authors:E. Farkas, B. Biró, K. Szabó, K. Veres, Zs. Csintalan, and R. Engel
The terricolous species Cladonia foliacea (Cladoniaceae, lichenised Ascomycota) widely distributed in open, dry lowland steppe and rocky mountain grassland vegetation in Europe was chosen as a potential test organism for ecological experiments, since their thalli are producing cortical solar radiation-protective and UV screening pigment dibenzofuran usnic acid and medullary secondary substance depsidone fumarprotocetraric acid. Significant seasonal differences were found in the amounts of lichen secondary metabolites analysed by HPTLC and HPLC-PDA between summer and winter collected thalli in sandy grassland area in Hungary. The concentrations of usnic acid varied between 7.34 and 15.52 mg/g in summer collected samples and 13.90 and 21.61 mg/g in winter collected ones. A comparable amount (11.61±0.29 mg/g) was measured in pulverised samples. The concentrations of fumarprotocetraric acid varied between 0.60 and 3.01 mg/g in summer collected samples and 2.26 and 5.81 mg/g in winter collected thalli. A comparable amount (2.45±0.21 mg/g) was found in pulverised samples. The range of concentration values is comparable with data known from lichens. A higher amount of usnic acid is produced in winter probably to ensure sufficient protection also for summer. The fumarprotocetraric acid content of the medulla might contribute to the solar irradiation reflecting role of the pale lower surface lobes turning upwards in dry condition.
Lichenes Delicati Exsiccati Editae of little, fine, special lichens is edited in honour of Antonín Vězda (1920–2008). The fifth fascicle of the exsiccate is consisted of 20 species of lichens and lichenicolous fungi and distributed to 12 lichen herbaria of the world. Collectors are K. Buaruang, D. Kalb, K. Kalb, G. E. Lee, L. Lőkös, A. Mertens, W. Polyiam, T. Pócs, W. Saipunkaew, D. Tang, N. Varga and E. Farkas.
Authors:T. Wirth, I. Fazekas, Cs. Schmidt, and J. Csiky
Between the years 2015–2018, 147 stands of Ficus carica L. was found out of cultivation in Baranya county. In 2008 presence of fig wasp (Blastophaga psenes L.) and caprificus individuals that are necessary for pollination, then for producing fertile seeds were detected in Pécs at first time. For germination tests fig seeds were collected from several different stands in Pécs and successful reproduction of the species was confirmed under in vitro conditions. According to former and recent observations (i.e. subspontaneous seedlings and/or successful in vitro germination of common fig seeds in 2010 and 2014) fig wasp may have occurred in Budapest and Máriagyűd too, at least in the last ten years. These results confirm that F. carica is an old ‘new’ casual or may be a naturalised neophyte element of the Hungarian flora. According to the new records the northernmost escaped individuals of F. carica was found at 47.475° N in the Carpathian Basin, 116.5 km N of the closest Slovenian stand in the latitudinal direction.
Authors:Zoltán Attila Köbölkuti, Klára Cseke, Attila Benke, Mátyás Báder, Attila Borovics, and Róbert Németh
Since Populus has veritable value as timber, plywood, pulp, and paper, genomic research should create the sound basis for further breeding toward desirable wood quality attributes.
Materials and methods
In this study, we addressed the need for a research methodology that initially identifies and then characterize candidate genes encoding enzymes with wood property phenotypic traits, toward the aim of developing a genomics-based breeding technology.
On 23 different poplar species/hybrid samples, we successfully amplified 55 primers designed on Populus trichocarpa L. Considering the number of polymorphic sites, out of 73,206 bp, 51 SNPs and 31 indel events were found. Non-synonymous single base mutations could be detected in number of 30, 21 out of 164 sequences were the number of minimum recombination events and 41 significant pairwise comparisons between loci could be detected.
Discussion and conclusion
Our results provide a roadmap for a future association genetic study between nucleotide diversity and precise evaluation of phenotype.
Authors:Gergely Sámuel Bartha, Gergő Tóth, Péter Horváth, Eszter Kiss, Nóra Papp, and Monika Kerényi
Several Aristolochia species were used as medicinal herb across Europe and in recent years, their antimicrobial activity has also been investigated.
Materials and methods
In this study, A. clematitis was selected to evaluate the aristolochic acids I and II (AA I and AA II) concentrations and the antimicrobial activity of methanol, hexane, butanol, and ethyl acetate extracts of the root, stem, leaf, root, and fruit. AA I and AA II contents were measured by a validated high-performance liquid chromatography–ultraviolet method.
Each fraction of the plant contained AA I and AA II and the root was found to have the highest contents of AA I (1.09%) and AA II (0.7454%). The minimum inhibitory concentrations of all extracts were determined by standard microdilution method. The fruit’s extracts showed the most efficient antimicrobial effect against both methicillin sensitive and resistant Staphylococcus aureus strains.
Correlation between the AA I and AA II concentrations and the antimicrobial effect was not found.
Due to its overall environmental impact, the residual dye in the wastewater from the synthetic dye manufacturing and textile industries is a global concern. The discharge contains a high content of pigments and other additives, possessing complex structures. As per the requirement for dyed clothing, dyestuff in the effluent is less susceptible to acids, bases, and oxygen. Thus, conventional physical and chemical methods are not always efficient in degrading the dyes. Some microorganisms growing in an area affected with textile effluent have the capability to utilize the dyes as a source of carbon or nitrogen or both. As a very clean, inexpensive, and sufficient alternative, bioremediation of textile wastewater using these microorganisms has gained major popularity. This review primarily centers the contribution of bacteria in this sector and the isolation of such bacteria from textile effluent. A secondary focus is discussing the factors which influence the performance by different bacteria.
Authors:Rubina Tu¨nde Szabó, Mária Kovács-Weber, Márta Erdélyi, Krisztián Balogh, Natasa Fazekas, Ákos Horváth, Miklós Mézes, and Balázs Kovács
Background and aims
The aim of this study was to verify that the comet assay can be used to investigate the DNA damaging effects of T-2 and HT-2 toxins in the liver of broiler chickens. The comet assay is a favorable genotoxic analysis because it is cheap, simple, and can be used in many organisms and different tissues.
Materials and methods
Male broiler chickens were fed with T-2/HT-2 toxins-contaminated diet for 14 days. The comet assay was successfully adapted to chicken liver cells, and the DNA damage was determined by a decrease in the comet parameter (DNA % in the tail) in the experimental groups.
The method of evaluation was found to be critical because DNA damage could not be detected exactly using the CometScore software, due to inaccurate separation of head and comet. However, this problem can be solved by visual evaluation.
In the case of the visual evaluation, each toxin-treated group differed significantly from the control group, indicating that the assay can be useful for the assessment of primary DNA damage caused by T-2/HT-2 toxins.
In this study, we analyzed gynandromorphs with female terminalia, to dissect mating-related female behaviors in Drosophila.
Materials and methods
We used gynandromorphs, experimentally modified wild-type (Oregon-R) females, and mutant females that lacked different components of the female reproductive apparatus.
Many of the gynandromorphs mated but did not expel the mating plug (MP). Some of these – with thousands of sperm in the uterus – failed to take up sperm into the storage organs. There were gynandromorphs that stored plenty of sperm but failed to release them to fertilize eggs. Expelling the MP, sperm uptake into the storage organs, and the release of stored sperm along egg production are separate steps occurring during Drosophila female fertility. Cuticle landmarks of the gynandromorphs revealed that while the nerve foci that control MP expelling and also those that control sperm uptake reside in the abdominal, the sperm release foci derive from the thoracic region of the blastoderm.
Discussion and conclusion
The gynandromorph study is confirmed by analyses of (a) mutations that cause female sterility: Fs(3)Avar (preventing egg deposition), Tm2gs (removing germline cells), and iab-4DB (eliminating gonad formation) and (b) by experimentally manipulated wild-type females: decapitated or cut through ventral nerve cord.