bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of selective antibacterial peptides by the Polarity Profile method.
Authors: Polanco Carlos , Buhse Thomas , Samaniego José Lino , Castañón-González Jorge Alberto
Published: February 2019

Antimicrobial peptides occupy a prominent place in the production of pharmaceuticals, because of their effective contribution to the protection of the immune system against almost all types of pathogens. These peptides are thoroughly studied by computational methods designed to shed light on their main functions. In this paper, we propose a computational approach, named the Polarity Profile method that represents an improvement to the former Polarity Index method. The Polarity Profile method is very effective in detecting the subgroup of antibacterial peptides called selective cationic amphipathic antibacterial peptides (SCAAP) that show high toxicity towards bacterial membranes and exhibit almost zero toxicity towards mammalian cells. Our study was restricted to the peptides listed in the antimicrobial peptides database (APD2) of December 19, 2012. Performance of the Polarity Profile method is demonstrated through a comparison to the former Polarity Index method by using the same sets of peptides. The efficiency of the Polarity Profile method exceeds 85% taking into account the false positive and/or false negative peptides.
Redox properties and prooxidant cytotoxicity of a neuroleptic agent 6,7-dinitrodihydroquinoxaline-2,3-dione (DNQX).
Authors: Šarlauskas Jonas , Nemeikaitė-Čėnienė Aušra , Misevičienė Lina , Krikštopaitis Kastis , Anusevičius Žilvinas , Čėnas Narimantas
Published: February 2019

In order to characterize the possible mechanism(s) of cytotoxicity of a neuroleptic agent 6,7-dinitrodihydroquinoxaline-2,3-dione (DNQX) we examined the redox properties of DNQX, and its mononitro- (NQX) and denitro- (QX) derivatives. The irreversible electrochemical reduction of the nitro groups of DNQX was characterized by the reduction peak potentials (Ep,7) of -0.43 V and -0.72 V vs. Ag/AgCl at pH 7.0, whereas NQX was reduced at Ep,7 = -0.67 V. The reactivities of DNQX and NQX towards the single-electron transferring enzymes NADPH:cytochrome P-450 reductase and NADPH:adrenodoxin reductase/adrenodoxin complex were similar to those of model nitrobenzenes with the single-electron reduction potential (E¹₇) values of -0.29 V - -0.42 V. DNQX and NQX also acted as substrates for two-electron transferring mammalian NAD(P)H:quinone oxidoreductase (DT-diaphorase). The cytotoxicity of DNQX in bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) was prevented by antioxidants and an inhibitor of NQO1, dicoumarol, and was enhanced by the prooxidant alkylating agent 1,3-bis(2-chloromethyl)-1-nitrosourea. A comparison with model nitrobenzene compounds shows that the cytotoxicity of DNQX and NQX reasonably agrees with the ease of their electrochemical reduction, and/or their reactivities towards the used enzymatic single-electron reducing systems. Thus, our data imply that the cytotoxicity of DNQX in FLK cells is exerted mainly through oxidative stress.
Norway spruce (Picea abies) genetic transformation with modified Cry3A gene of Bacillus thuringiensis.
Authors: Bříza Jindřich , Pavingerová Daniela , Vlasák Josef , Niedermeierová Hana
Published: February 2019

Modified versions of the Cry3A gene of Bacillus thuringiensis (Bt) were transferred into Norway spruce (Picea abies). Both the biolistic approach and Agrobacterium tumefaciens mediated procedure were employed for transformation of embryogenic tissue (ET) cultures. The latter method proved to be more efficient yielding 70 transgenic embryogenic tissue lines compared with 18 lines obtained by biolistics. The modified Cry3A genes were driven by a 35S promoter and the nptII screenable selection marker gene was used in all vectors. The transgenic ETs were molecularly characterized and converted into mature somatic embryos. Germinating embryos formed plantlets which were finally planted into perlite and their Cry3A gene transcription activities were demonstrated by RT-PCR.
Detection and differentiation of Newcastle disease virus and influenza virus by using duplex real-time PCR.
Authors: Nidzworski Dawid , Wasilewska Edyta , Smietanka Krzysztof , Szewczyk Bogusław , Minta Zenon
Published: February 2019

Newcastle disease virus (NDV), member of the Paramyxoviridae family and avian influenza virus (AIV), member of the Orthomyxoviridae family, are two main avian pathogens causing serious economic problems in poultry health. Both are enveloped, single-stranded, negative-sense RNA viruses and cause similar symptoms, ranging from sub-clinical infections to severe diseases, including decrease in egg production, acute respiratory syndrome, and high mortality. Similar symptoms hinder the differentiation of infection with the two viruses by standard veterinary procedures like clinical examination or necropsy. To overcome this problem, we have developed a new duplex real-time PCR assay for the detection and differentiation of these two viruses. Eighteen NDV strains, fourteen AIV strains, and twelve other (negative control) strains viruses were isolated from allantoic fluids of specific pathogen-free (SPF), embryonated eggs. Four-weeks-old SPF chickens were co-infected with both viruses (NDV - LaSota and AIV - H7N1). Swabs from cloaca and trachea were collected and examined. The results obtained in this study show that by using duplex real-time PCR, it was possible to detect and distinguish both viruses within less than three hours and with high sensitivity, even in case a bird was co-infected. Additionally, the results show the applicability of the real-time PCR assay in laboratory practice for the identification and differentiation of Newcastle disease and influenza A viruses in birds.
Single dose of green tea extract decreases lipid digestion and absorption from a test meal in humans.
Authors: Walkowiak Jarosław , Bajerska Joanna , Kargulewicz Angelika , Lisowska Aleksandra , Siedlerski Gwidon , Szczapa Tomasz , Kobelska-Dubiel Natalia , Grzymisławski Marian
Published: February 2019

Green tea is known worldwide for its high content of polyphenolic compounds and multifactorial beneficial effects on human health. The role of green tea as an inhibitor of lipid hydrolysis is widely discussed. The aim of the study was to assess the influence of green tea extract on lipid digestion and absorption.
The study comprised 32 healthy volunteers aged 23 to 30 years with normal exocrine pancreatic function. In all subjects (13)C-labelled mixed triglyceride breath test was performed twice with and without green tea extract ingestion. Cumulative percentage dose recovery was considered to reflect digestion and absorption of lipids. Values are expressed as medians and 1st-3rd quartile distribution.
In all subjects, cumulative percentage dose recovery values were normal in a placebo test (36.8% ). These results were significantly higher (p=0.021) than those obtained in green tea extract test (28.8% ). Results of six tests with GTE were abnormal.
Single dose of green tea extract taken with a test meal decreases lipid digestion and absorption in humans.
Alternative 3' acceptor site in the exon 2 of human PAX8 gene resulting in the expression of unknown mRNA variant found in thyroid hemiagenesis and some types of cancers.
Authors: Szczepanek-Parulska Ewelina , Szaflarski Witold , Piątek Katarzyna , Budny Bartłomiej , Jaszczyńska-Nowinka Karolina , Biczysko Maciej , Wierzbicki Tomasz , Skrobisz Jerzy , Zabel Maciej , Ruchała Marek
Published: February 2019

PAX8 gene encodes one of the transcription factors engaged in the regulation of proper development of thyroid gland as well as Müllerian and renal/upper urinary tracts. So far, six alternatively spliced transcripts were reported, however, sequences of only four were deposited in the NCBI database. Here, we evaluate a fragment of a novel variant of PAX8 mRNA formed by an alternative 3' acceptor site located in the second exon. The molecular outcome encompasses extension of the 5' untranslated region of exon two by 97 nucleotides as is evident from mRNA. This new insert may impair binding of mRNA to the ribosome and in consequence significantly decrease expression of the PAX8 protein. Here, we show for the first time that the novel insert in exon two might be associated with congenital thyroid hemiagenesis and influence development of different types of cancer.
Design, expression and characterization of a highly stable tetratricopeptide-based protein scaffold for phage display application.
Authors: Petters Edyta , Krowarsch Daniel , Otlewski Jacek
Published: February 2019

Tetratricopeptide repeat (TPR) is a structural motif mediating variety of protein-protein interactions. It has a high potential to serve as a small, stable and robust, non-immunoglobulin ligand binding scaffold. In this study, we showed the consensus approach to design the novel protein called designed tetratricopeptide repeat (dTPR), composed of three repeated 34 amino-acid tetratricopeptide motifs. The designed sequence was efficiently overexpressed in E. coli and purified to homogeneity. Recombinant dTPR is monomeric in solution and preserves its secondary structure within the pH range from 2.0 to 11.0. Its denaturation temperature at pH 7.5 is extremely high (104.5°C) as determined by differential scanning calorimetry. At extreme pH values the protein is still very stable: denaturation temperature is 90.1°C at pH 2.0 and 60.4°C at pH 11. Chemical unfolding of the dTPR is a cooperative, two-state process both at pH 7.5 and 2.0. The free energy of denaturation in the absence of denaturant equals to 15.0 kcal/mol and 13.5 kcal/mol at pH 7.5 and 2.0, respectively. Efficient expression and extraordinary biophysical properties make dTPR a promising framework for a biotechnological application, such as generation of specific ligand- binding molecules.
Microflora of the selected water reservoirs in Swietokrzyskie Voivodship.
Authors: Adamus-Bialek Wioletta , Karwacka Karolina , Bak Lukasz
Published: February 2019

One of the important environmental issues is the quality of surface waters in the world. Poland belongs to countries with a low quality of the inland waters. The sanitary condition of the five water reservoirs of south-east Poland was analyzed. Water and sediment samples were incubated on the selective and/or differential media. High concentrations of many common and pathogenic microbial indicators were shown in those samples. Those reservoirs are used by people, especially during summer. Because of the high epidemiological risk, detailed analysis of all inland waters should be performed routinely.
Phosphorus cycle - possibilities for its rebuilding.
Authors: Gorazda Katarzyna , Wzorek Zbigniew , Tarko Barbara , Nowak Anna K , Kulczycka Joanna , Henclik Anna
Published: February 2019

The rebuilding of the phosphorus cycle can be performed with the use of both biotechnology and chemical technology. This paper presents a review of the phosphorus cycle and the different approaches that can be taken to the recovery of phosphorus from phosphate-rich waste. Critical issues in the phosphorus cycle are also discussed. Methods for the recovery of phosphorus form sewage sludge ash are widely explored and divided into two groups: wet extraction methods and thermochemical methods. Laboratory-scale methods are described, as well as proposed industrial technologies, with particular regard to the possibilities for their implementation in Poland. Phosphorus recovery methods from SSA (sewage sludge ash) in our country seems to be promising due to the increasing number of sewage sludge incineration plants, which could easily supply ash to future recovery installations. For the effective recovery of P from sewage sludge ash, it is essential to make the right choice in determining the appropriate method to use with respect to the particular properties of the ash composition available. A patented method of phosphorus recovery by acid extraction methods, developed by Cracow University of Technology, results in an efficiency of 80-96% for phosphorus recovery. 3000 to 4000 tons of phosphorus per year can be recycled and introduced back into the environment, that covers around 7% of the total amount of phosphorus ore imported into Poland between 2008 and 2009.
Phenolics, antioxidative activity and characterization of anthocyanins in berries of blackcurrant interspecific hybrids.
Authors: Anisimovienė Nijolė , Jankauskienė Jurga , Jodinskienė Milda , Bendokas Vidmantas , Stanys Vidmantas , Šikšnianas Tadeušas
Published: February 2019

The interest in phenols and anthocyanins has increased due to their antioxidant properties and to their potential usage as dietary antioxidants in human nutrition. Total phenols and anthocyanin content, composition and stability in berry extracts of blackcurrant interspecific hybrids, and antioxidative activity of extracts was evaluated. Berries of interspecific hybrids accumulated 530 to 614 mg 100 g(-1) FW of total phenolic compounds, while 621 mg 100 g(-1) FW of phenolics was established in berries of control Ribes nigrum cultivar 'Ben Tirran'. 'Ben Tirran' berries accumulated 444 mg 100 g(-1) FW of anthocyanins and higher amount was identified in berries of interspecific hybrids No. 11-13 (R. nigrum × R. petraeum) and No. 57 (R. nigrum × R. aureum), 522 and 498 mg100 g(-1) FW respectively. Berry extracts of hybrid No. 11-13 distinguished by the highest antioxidative activity (80%) and it was higher than antioxidant activity of 'Ben Tirran' (70%). Antioxidative activity of all tested berry extracts (70-80%) was twice higher compared to synthetic antioxidant BHT (39%). However correlation between phenolics or total anthocyanin content and antioxidative activity degree was not established. Amount of cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside in berries of hybrids No. 57, No. 11-13 and No. 8 ((R. nigrum × R. americanum) × (R. nigrum × R. americanum)) was higher than in berries of 'Ben Tirran'. It was established that cyanidins are more stable anthocyanins in all studied temperature and irradiation conditions. Therefore interspecific hybrids No. 57 and No. 11-13 were the most agronomically valuable hybrids.
The kinetic reduction of Cr(VI) by yeast Saccharomyces cerevisiae, Phaffia rhodozyma and their protoplasts.
Authors: Chwastowski Jarosław , Kołoczek Henryk
Published: February 2019

Chromium in the sixth oxidation state may easily penetrate cellular membranes via non-specific sulfate transporters due to its tetrahedral symmetry (high similarity to SO4(2-) and HPO4(2-)). This feature makes chromium a toxic and hazardous pollutant responsible for the deterioration of midland water quality. The aim of the study was to evaluate the capacity of two yeast species - Saccharomyces cerevisiae and Phaffia rhodozyma - and their protoplasts to reduce Cr(VI) to lower oxidation states. The study also deals with the behavior of the yeasts upon the presence of elevated sulfate ions as a competitive inhibitor of chromate transport by the sulfate transporters. The chromate-reducing activities were monitored by determination of Cr(V) free radical form with the use of L-band (1.2 GHz) EPR (electron paramagnetic resonance) spectroscopy. It was observed that both of the studied yeast strains exhibited the ability to reduce Cr(VI) applied at 4 mM. The cells of P. rhodozyma showed about 3.5 times higher reduction than S. cerevisiae. The reduction efficiency was significantly improved when the protoplasts of both strains were used and reached 100% in the first 10 minutes of the reduction process which suggests that the cellular wall may have a notable influence on the uptake and/or inhibition of chromium reduction process. The reduction effect of P. rhodozyma cells and protoplasts may be associated with the more sufficient production of metabolites (such as glutathione and cysteine), which may also be responsible for the increased tolerance of the strain towards high concentrations of toxic chromium.
Prp4 kinase is required for proper segregation of chromosomes during meiosis in Schizosaccharomyces pombe.
Authors: Pozgajova Miroslava , Cipak Lubos , Trakovicka Anna
Published: February 2019

Chromosome segregation during meiosis is a complex process, which leads to production of four haploid gametes from two precursor cells. Reversible phosphorylation of proteins plays a crucial role in this process. The Schizosaccharomyces pombe Prp4 is an essential serine/threonine protein kinase, which belongs to the Clk/Sty family. To study the role of Prp4 in meiosis, we analysed chromosome segregation in a strain carrying conditional analog-sensitive allele of Prp4 protein kinase (prp4-as2). Our data show, that Prp4 protein kinase plays important role in chromosome segregation during meiosis, as revealed by enhanced missegregation of chromosomes in prp4-as2 mutant cells.
Supporting the process of removing humic substances on activated carbon.
Authors: Olesiak Paulina , Stępniak Longina
Published: February 2019

This study is focused on biosorption process used in water treatment. The process has a number of advantages and a lot of research has been done into its intensification by means of ultrasonic modification of solutions. The study carried out by the authors leads to the conclusion that sonication of organic solutions allows for extension of the time of operation of carbon beds. For the analysis of the results obtained during the sorption of humic substances (HS) from the solution dependencies UV/UV₀ or DOC/DOC₀ were used. In comparative studies the effectiveness of sorption and sonosorption (UV/UV₀) shows that the share of ultrasounds (US) is beneficial for extension of time deposit, both at a flow rate HS solution equal to 1 m/h and 5 m/h. Analysis of the US impact sorption on HS sorption in a biological fluidized bed, both prepared from biopreparat and the activated sludge confirms the higher efficiency compared to sonobiosorption than biosorption. These results confirm the degree of reduction UV₂₅₄/UV₀ and DOC/DOC₀ for the same processes. EMS index also confirms the improvement of HSbiodegradation by sludge microorganisms.
Hypothetical glycerol pathways of newly isolated strains capable of 1,3-propanediol production.
Authors: Leja Katarzyna , Samul Dorota , Drożdżyńska Agnieszka , Myszka Kamila , Juzwa Wojciech , Pawlicka Joanna , Czaczyk Katarzyna
Published: February 2019

Study presented here demonstrates the ability of three newly isolated strains, obtained from environmental probes (manure, bottom sediment, and food waste) and identified as Clostridium bifermentans, Clostridium butyricum, and Hafnia alvei, to synthesize 1,3-propanediol (1,3-PD), organic acids (such as lactic, acetic, fumaric, succinic, and butyric acids), and ethanol from glycerol. The production of 1,3-PD as well as the glycerol pathways in C. bifermentans and H. alvei cells have not been investigated and described yet by others. Moreover, there is no data in the available literature on the products of glycerol utilization by H. alvei and there is only some incoherent data (mainly from the first half of the twentieth century) about the ability of C. bifermentans to carry out glycerol degradation. Additionally, this study presents complete hypothetical glycerol pathways and the basic fermentation kinetic parameters (such as yield and productivity) for both strains as well as for the newly isolated C. butyricum strain.
The strategy of fusion genes construction determines efficient expression of introduced transcription factors.
Authors: Adamus Tomasz , Konieczny Paweł , Sekuła Małgorzata , Sułkowski Maciej , Majka Marcin
Published: February 2019

The main goal in gene therapy and biomedical research is an efficient transcription factors (TFs) delivery system. SNAIL, a zinc finger transcription factor, is strongly involved in tumor, what makes its signaling pathways an interesting research subject. The necessity of tracking activation of intracellular pathways has prompted fluorescent proteins usage as localization markers. Advanced molecular cloning techniques allow to generate fusion proteins from fluorescent markers and transcription factors. Depending on fusion strategy, the protein expression levels and nuclear transport ability are significantly different. The P2A self-cleavage motif through its cleavage ability allows two single proteins to be simultaneously expressed. The aim of this study was to compare two strategies for introducing a pair of genes using expression vector system. We have examined GFP and SNAI1 gene fusions by comprising common nucleotide polylinker (multiple cloning site) or P2A motif in between them, resulting in one fusion or two independent protein expressions respectively. In each case transgene expression levels and translation efficiency as well as nuclear localization of expressed protein have been analyzed. Our data showed that usage of P2A motif provides more effective nuclear transport of SNAIL transcription factor than conventional genes linker. At the same time the fluorescent marker spreads evenly in subcellular space.
Evaluation of the effects of antibiotics on cytotoxicity of EGFP and DsRed2 fluorescent proteins used for stable cell labeling.
Authors: Zakrzewska Karolina Ewa , Samluk Anna , Pluta Krzysztof Dariusz , Pijanowska Dorota Genowefa
Published: February 2019

The use of fluorescent markers has proven to be an attractive tool in biological imaging. However, its usefulness may be confined by the cytotoxicity of the fluorescent proteins. In this article, for the first time, we have examined an influence of the antibiotics present in culture medium on cytotoxicity of the EGFP and DsRed2 markers used for whole-cell labeling. Results showed that doxycycline negatively affected albumin synthesis in DsRed2-expressing hepatoma cells, and that both hepatoma cells and human skin fibroblasts, labeled with this protein, were characterized by the lowered growth rates. Thus, the cytotoxic effect of fluorescent markers depends on both protein used for cell labeling and on growth conditions that may cause cell stress.
Native nucleic acid electrophoresis as an efficient alternative for genotyping method of influenza virus.
Authors: Pajak Beata , Lepek Krzysztof
Published: February 2019

Influenza viruses are the worldwide major causative agents of human and animal acute respiratory infections. Some of the influenza subtypes have caused epidemics and pandemics among humans. The varieties of methods are available for the rapid isolation and identification of influenza viruses in clinical and environmental samples. Since nucleic acids amplification techniques such as RT-PCR have been adapted, fast and sensitive influenza type and subtype determination is possible. However, in some ambiguous cases other, more detailed assay might be desired. The genetic material of influenza virus is highly unstable and constantly mutates. It is known that single nucleotide polymorphisms (SNPs) results in resistance to commercially available anti-viral drugs. The genetic drift of the virus could also result in weakening of immune response to infection. Finally, in a substantial number of patients co-infection with various virus strains or types has been confirmed. Although the detection of co-infection or presence of minor genetic variants within flu-infected patients is not a routine procedure, a rapid and wide spectrum diagnostics of influenza virus infections could reveal an accurate picture of the disease and more importantly, is crucial for choosing the appropriate therapeutics and virus monitoring. Herein we present the evidences that native gel electrophoresis and MSSCP--a method based on multitemperature single strand conformation polymorphism could furnish a useful technique for minor variants, which escape discovery by conventional diagnostic assays.
Plant expression systems for production of hemagglutinin as a vaccine against influenza virus.
Authors: Redkiewicz Patrycja , Sirko Agnieszka , Kamel Katarzyna Anna , Góra-Sochacka Anna
Published: February 2019

Many examples of a successful application of plant-based expression systems for production of biologically active recombinant proteins exist in the literature. These systems can function as inexpensive platforms for the large scale production of recombinant pharmaceuticals or subunit vaccines. Hemagglutinin (HA) is a major surface antigen of the influenza virus, thus it is in the centre of interests of various subunit vaccine engineering programs. Large scale production of recombinant HA in traditional expression systems, such as mammalian or insect cells, besides other limitations, is expensive and time-consuming. These difficulties stimulate an ever-increasing interest in plant-based production of this recombinant protein. Over the last few years many successful cases of HA production in plants, using both transient and stable expression systems have been reported. Various forms of recombinant HA, including monomers, trimers, virus like particles (VLPs) or chimeric proteins containing its fusion with other polypeptides were obtained and shown to maintain a proper antigenicity. Immunizations of animals (mice, ferrets, rabbits or chickens) with some of these plant-derived hemagglutinin variants were performed, and their effectiveness in induction of immunological response and protection against lethal challenge with influenza virus demonstrated. Plant-produced recombinant subunit vaccines and plant-made VLPs were successfully tested in clinical trials (Phase I and II) that confirmed their tolerance and immunogenicity.
Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris.
Authors: Kopera Edyta , Dwornyk Angela , Kosson Piotr , Florys Katarzyna , Sączyńska Violetta , Dębski Janusz , Cecuda-Adamczewska Violetta , Szewczyk Bogusław , Zagórski-Ostoja Włodzimierz , Grzelak Krystyna
Published: February 2019

The A/swan/Poland/305-135V08/2006 (H5N1-subtype) hemagglutinin (HA) gene was cloned and expressed in yeast Pichia pastoris (P. pastoris). The HA cDNA lacking the C-terminal transmembrane anchor-coding sequence was fused to an α-factor leader peptide and placed under control of the methanol-inducible P. pastoris alcohol oxidase 1 (AOX1) promoter. Two P. pastoris strains: SMD 1168 and KM 71 were used for protein expression. Recombinant HA protein was secreted into the culture medium reaching an approximately 15 mg/L (KM 71 strain). Fusion protein with a His6 tag was purified to homogeneity in one step affinity chromatography. SDS-PAGE and MS/MS analysis indicated that the protein is cleaved into HA1 and HA2 domains linked by a disulfide bond. Analysis of the N-linked glycans revealed that the overexpressed HA is fully glycosylated at the same sites as the native HA in the vaccine strain. Immunological activity of the hemagglutinin protein was tested in mice, where rHA elicited a high immune response.
Genetic diversity of F1 and F2 interspecific hybrids between dwarf birch (Betula nana L.) and Himalayan birch (B. utilis var. jacquemontii (Spach) Winkl. 'Doorenbos') using RAPD-PCR markers and ploidy analysis.
Authors: Czernicka Małgorzata , Pławiak Jarosław , Muras Piotr
Published: February 2019

Crosses between Betula nana and B. utilis 'Doorenbos' were undertaken in order to obtain interspecific hybrids which could be characterized by wide spreading stems, strong branching habit, decorative clear white bark and an interesting shape of purple leaves. The research purpose was to examine genetic diversity of the 16 F1 and F2 putative progenies by using the RAPD-PCR method and the ploidy analysis. A total of 242 RAPD markers were scored with 24 primers and 220 (90.9%) polymorphic bands were found. In the NJ dendrogram, cluster I consisted of the female parent--B. nana and 12 hybrids and cluster II grouped the male parent--B. utilis 'Doorenbos' with 4 hybrids (F2/2, F1/8, F1/7 and F2/1). The 2-D scaling by PCoA was in agreement with the similarity index, i.e. two hybrids (F1/8, F2/2) grouped with the male parent while others with female parent. Classification of the hybrid plants by chromosome counting demonstrated that 13 hybrids were confirmed with accurate chromosome counts as being diploid (2n=2x=28) and 3 plants (F1/7, F1/8, F2/2) as triploid with 42 chromosomes.
Ex-translational function of tRNAs and their fragments in cancer.
Authors: Mleczko Anna M , Celichowski Piotr , Bąkowska-Żywicka Kamilla
Published: February 2019

Transfer RNA (tRNA) molecules are most commonly known as the molecular amino acids carriers and also because of the role they play in a protein biosynthesis process. However, tRNA biology has revealed stupendous levels of many unexpected discoveries that put a new light on tRNA function in different processes besides translation, like apoptosis or cancer development. In recent years various species of RNAs have been found differentially expressed in different types of cancer. In this review we focus our attention on tRNAs as well as on tRNA-derived small RNAs ex-translational functions in human cells in oncogenesis and oncobiology.
Maximal oxygen uptake is associated with allele -202 A of insulin-like growth factor binding protein-3 (IGFBP3) promoter polymorphism and (CA)n tandem repeats of insulin-like growth factor IGF1 in Caucasians from Poland.
Authors: Gronek Piotr , Holdys Joanna , Kryściak Jakub , Wieliński Dariusz , Słomski Ryszard
Published: February 2019

Physical fitness is a trait determined by multiple genes, and its genetic basis is modified by numerous environmental factors. The present study examines the effects of the (CA)n tandem repeats polymorphism in IGFI gene and SNP Alw21I restriction site -202 A>C polymorphism in IGF1BP3 on VO2max--a physiological index of aerobic capacity of high heritability. The study sample consisted of 239 (154 male and 85 female) students of the University School of Physical Education in Poznań and athletes practicing various sports, including members of the Polish national team. An association was found between -202 A/C polymorphism of IGFBP3 gene with VO2max in men. Higher VO2max values were attained by men with CC genotype, especially male athletes practicing endurance sports and sports featuring energy metabolism of aerobic/anaerobic character. A statistically significant influence of allele 188 and genotype 188/188 of tandem repeats (CA)n polymorphism of IGF1 gene on VO2max was found in women. Also, lower values of maximal oxygen uptake were noted in individuals with allele 186 or genotype 186/186, and higher VO2max values in athletes with allele 194.
The pH optimum of native uracil-DNA glycosylase of Archaeoglobus fulgidus compared to recombinant enzyme indicates adaption to cytosolic pH.
Authors: Knævelsrud Ingeborg , Kazazic Sabina , Birkeland Nils-Kåre , Bjelland Svein
Published: February 2019

Uracil-DNA glycosylase of Archaeoglobus fulgidus (Afung) in cell extracts exhibited maximal activity around pH 6.2 as compared to pH 4.8 for the purified recombinant enzyme expressed in Escherichia coli. Native Afung thus seems to be adapted to the intracellular pH of A. fulgidus, determined to be 7.0±0.1. Both recombinant and native Afung exhibited a broad temperature optimum for activity around 80°C, reflecting the A. fulgidus optimal growth temperature of 83°C. Adaption to the neutral conditions in the A. fulgidus cytoplasm might be due to covalent modifications or accessory factors, or due to a different folding when expressed in the native host.
Response of the pea roots defense systems to the two-element combinations of metals (Cu, Zn, Cd, Pb).
Authors: Malecka Arleta , Piechalak Aneta , Zielińska Barbara , Kutrowska Agnieszka , Tomaszewska Barbara
Published: February 2019

The presence of the single metals (Cd, Pb, Cu, Zn) induces ROS (reactive oxygen species) production and causes oxidative stress in plants. While applied in two-element combinations, trace metals impact organisms in a more complex way. To assess the resultant effect we treated the pea grown hydroponically with the trace metals in variants: CuPb, CuCd, CuZn, PbCd, ZnPb, ZnCd in concentrations of 25 µM for each metal ion. Abiotic stress inhibited root elongation growth, decreased biomass production, induced changes in root colour and morphology. It changed rate of ROS production, malondialdehyde content, increased activity and altered gene expression of defence enzymes (superoxide dysmutase, catalase, ascorbate peroxidase, glutathione reductase, γ-glutamylcysteine synthetase).
mRNA Regulatory elements and bacterial virulence.
Authors: Pawlikowska-Warych Małgorzata , Deptuła Wiesław
Published: February 2019

Pathogenic bacteria cause many diseases, some of which are fatal. For researchers, it is a challenge to understand bacterial mechanisms of pathogenicity, including their virulence pathways regulated by RNA. This work presents data on the mechanisms of regulation and expression of several virulence factors coded by RNA, namely 5' UTR fragments, riboswitches and small non-coding RNA (sRNA).
Molecular characterization of Capra hircus lysosomal α-mannosidase and potential mutant site for the therapy of locoweed poisoning.
Authors: Xiangya Kong , Jiangye Zhang , Ying Wu , Jianfei Li , Qinfan Li
Published: February 2019

Lysosomal α-Mannosidase (LAM) belongs to the glycoside hydrolyzing enzymes family 38 and is involved in the biosynthesis and turnover of N-linked glycoproteins process. Locoweeds, which contain swainsonine (SW) that inhibits LAM, are the main poisoning plants in many regions of the world, and thereby resulting in animal poisoning or even death. Based on regions of protein sequence conservation between LAM from Bos taurus and Homo sapiens, we cloned cDNA encoding Capra hircus LAM (chLAM). Expression of cDNA in Pichia pastoris resulted in the secretion of aLAM activity into the culture medium. The recombinant chLAM was activated 1.6 and 1.2-fold with Zn(2+) and Ca(2+), respectively. By homology modeling, molecular docking and mutant analysis, we obtained the probable binding modes of SW at the allosteric sites of chLAM, and the potential mutant sites for the resistance to SW. Prediction of SW sensitivity to A28 W/G, D58 Y/G mutant chLAM is lower than wild type chLAM. The obtained results lead to a better understanding of not only interactions between substrate/SW and chLAM, but also of a potential strategy for a novel therapy for locoweed poisoning.
Comparison of fecal pyruvate kinase isoform M2 and calprotectin in assessment of pediatric inflammatory bowel disease severity and activity.
Authors: Czub Elzbieta , Nowak Jan K , Szaflarska-Poplawska Anna , Grzybowska-Chlebowczyk Urszula , Landowski Piotr , Moczko Jerzy , Adamczak Daria , Mankowski Przemyslaw , Banasiewicz Tomasz , Plawski Andrzej , Walkowiak Jaroslaw
Published: February 2019

Accurate assessment of inflammatory bowel disease (IBD) activity is the cornerstone of effective therapy. Fecal M2 isoform of pyruvate kinase (M2-PK) and fecal calprotectin (FC) are noninvasive markers of mucosal inflammation in IBD. The aim of this study was to compare performance of M2-PK and FC in assessment of pediatric ulcerative colitis (UC) and Crohn's disease (CD) severity and activity.
121 patients with IBD, including 75 with UC and 46 with CD were recruited. Control group consisted of 35 healthy children (HS). Patients were assigned to groups depending on disease severity and activity. M2-PK and calprotectin concentration were determined in stool samples using ELISA. Areas under receiver operating characteristic curves (AUC) for FC and M2-PK with cut-off level at which M2-PK specificity was matching FC specificity were calculated and compared.
Performance of M2-PK at identifying patients with IBD, UC and CD among HS was inferior to FC. The differences in AUC were respectively: -0.10 (95% confidence interval [CI] [-0.13-(-0.06)], p
Effect of nutrient and stress factors on polysaccharides synthesis in Proteus mirabilis biofilm.
Authors: Moryl Magdalena , Kaleta Aleksandra , Strzelecki Kacper , Różalska Sylwia , Różalski Antoni
Published: February 2019

The extracellular matrix in biofilm consists of water, proteins, polysaccharides, nucleic acids and phospholipids. Synthesis of these components is influenced by many factors, e.g. environment conditions or carbon source. The aim of the study was to analyse polysaccharides levels in Proteus mirabilis biofilms after exposure to stress and nutritional conditions. Biofilms of 22 P. mirabilis strains were cultivated for 24, 48, 72 hours, 1 and 2 weeks in tryptone soya broth or in modified media containing an additional amount of nutrients (glucose, albumin) or stress factors (cefotaxime, pH 4, nutrient depletion). Proteins and total polysaccharides levels were studied by Lowry and the phenol-sulphuric acid methods, respectively. Glycoproteins levels were calculated by ELLA with the use of selected lectins (WGA and HPA). For CLSM analysis dual fluorescent staining was applied with SYTO 13 and WGA-TRITC. In optimal conditions the levels of polysaccharides were from 0 to 442 μg/mg of protein and differed depending on the strains and cultivation time. The agents used in this study had a significant impact on the polysaccharides synthesis in the P. mirabilis biofilm. Among all studied components (depending on tested methods), glucose and cefotaxime stimulated the greatest production of polysaccharides by P. mirabilis strains (more than a twofold increase). For most tested strains the highest amounts of sugars were detected after one week of incubation. CLSM analysis confirmed the overproduction of N-acetyloglucosamine in biofilms after cultivation in nutrient and stress conditions, with the level 111-1134%, which varied depending on the P. mirabilis strain and the test factor.
Thermodynamic aspects of the self-assembly of DsrA, a small noncoding RNA from Escherichia coli.
Authors: Geinguenaud Frédéric , Gesson Maeva , Arluison Véronique
Published: February 2019

DsrA is an Escherichia coli small noncoding RNA that acts by base pairing to some mRNAs in order to control their translation and turnover. It was recently shown that DsrA is able to self-associate in a way similar to DNA and to build nanostructures. Although functional consequence of this RNA self-assembly in vivo is not yet understood, the formation of such an assemblage more than likely influences the noncoding RNA function. We report here for the first time the thermodynamic basis of this natural RNA self-assembly. In particular we show that assembling of the ribonucleic acid is enthalpy driven and that the versatility of the RNA molecule is important for the polymerisation; indeed, an equivalent DNA sequence is unable to make a nanoassembly. The origin of the difference is discussed herein.
Activation of MAP kinase pathways in Galleria mellonella infected with Bacillus thuringiensis.
Authors: Wojda Iwona , Koperwas Konrad , Jakubowicz Teresa
Published: February 2019

We followed changes in the level of phospho-MAP kinases in the greater wax moth Galleria mellonella after infection with Bacillus thuringiensis. We observed an enhanced level of phosphorylated p38 and JNK in fat bodies of the infected larvae. In hemocytes, injection of B. thuringiensis caused the highest increase in phospho-JNK, however, all pathways were activated after aseptic injection. We report that Galleria mellonella larvae exposed to heat shock before infection showed an enhanced level of phosphorylated JNK in fat body. This finding is relevant in the light of our previous reports, which submit evidence that pre-shocked animals are more resistant to infection.
IRAP-analysis for evaluating the genetic stability of endemic and endangered species of the Western Caucasus flora in the collection in vitro
Authors: Suprun I. I. , Malyarovskaya V. I. , Stepanov I. V. , Samarina L. S.
Published: February 2019

The characterization of genetic diversity is one of the main components of the genetic resources collection and management. Molecular markers are the most effective tool for characterizing and assessing genetic diversity in plant collections. IRAP (inter-retrotransposons amplified polymorphism) markers have proven to be some of the most effective for characterizing and evaluating germplasm, confirming the genetic fidelity of in vitro preserved cultivars and species. In this regard, the aim of this work is to test several IRAP primers to identify genetic polymorphism and study the genetic fidelity of three rare and endemic flora species of the Western Caucasus during in vitro conservation. Approbation of 16 IRAP-primers on the investigated species was carried out for Eryngium maritimum L., Galanthus woronowii Losinsk. and Campanula sclerophylla Kolak. The results made it possible to select the most efficient of them for genetic fidelity analysis of micropropagated plants. Out of 16 IRAP primers 8 amplified PCR products in Eryngium maritimum. In Galanthus woronowii as well 8 of 16 IRAP primers resulted in the amplification with the number of DNA fragments ranging from 2 to 12. In Campanula sclerophylla 9 of 16 IRAP primers amplified 1 to 11 fragments, depending on the marker. The results of the genotyping of regenerants were compared with data on stock in situ plants, which were the source of explants for in vitro conservation. In total, 60 regenerants for each species of the natural flora of the Western Caucasus were involved in the study. The results obtained demonstrated no genetic changes of the regenerants in all the studied species. These results were confirmed using ISSR analysis of an extended sample set of microplants for each species. The results obtained can serve as evidence of a low probability of genetic disorders during in vitro propagation and conservation of the species Eryngium maritimum L., Galanthus woronowii Losinsk. and Campanula sclerophylla Kolak.
Genotypic variability of Pinus sylvestris L. on the drought-resistance attribute
Authors: Amineva E. Yu. , Gureev A. P. , Tabatskaya T. M. , Mashkina O. S. , Popov V. N.
Published: February 2019

There was an increase and intensification of droughts in connection with global climate change in recent decades. Not all plant organisms are able to adapt to changing environmental conditions. Therefore, the question of stressresistant (drought-resistant) genotypes selection for breeding is quite urgent. This problem also concerns forest tree plants, including Scots pine, which is one of the main foresters in the Voronezh region. The results of vegetative and generative sphere analysis of individual Scotch pine trees with the help of biotechnology, molecular-genetic and cytogenetic method are given in this study. The possibility of applying the tissue culture method in vitro for testing initial plants for stress resistance, including drought resistance, is explained by the interconnection of cells, tissues and the whole plant properties. It is shown that the cytogenetic characteristics of the seed progeny and the indicators of callusogenic reactions do not always coincide in different genotypes: in some cases energy resources are spent on ontogeny protection, while, in other cases, to reproductive function maintain. There are trees, in which the state the generative sphere in arid years is at the level of optimal years, and their callus cultures reaction remains unchanged even under simulated conditions of drought. Based on the results obtained for the selection of Scots pine drought-resistant genotypes we suggest applying a system of criteria characterizing both the ability of the vegetative sphere to survive in drought conditions on the basis of the tissue culture method in vitro (callus tissue formation speed, its viability, frequency of callusogenesis), and the state of the generative sphere with the help of cytogenetic analysis of seed progeny (frequency of mitosis pathologies, the proportion of cells with micronuclei, mitotic activity). The expediency of applying the biotechnological approach was proved by analysis of the gene expression level of stress proteins: the level of AbaH gene expression correlates greatly with the indicator of the viability of callus cultures, including ones on a nutrient medium with an additional stress agent (NaCl). Trees that can be assigned to drought-resistant ones, according to the results of the analysis, should be recommended for use in breeding.
Challenges of in vitro conservation of Сitrus germplasm resources
Authors: Gorshkov V. M. , Samarina L. S. , Kulyan R. V. , Malyarovskaya V. I. , Ryndin A. V. , Rakhmangulov R. S. , Orlov Y. L.
Published: February 2019

The main problems of establishment a slow growth in vitro collection of citrus and other tree crops cultivars are high degree of fungal contamination of bud explants and low growth potential of shoots. In this regard, the aim of current research is to assess the efficiency of decontamination procedure and the possibility of tissue culture initiation and slow growth conservation of valuable lemon cultivars. The best results of surface sterilization were obtained using immersion solutions of 0.3 % Veltolen – 25 minutes or 10 % Domestos – 25–30 minutes. In these treatments, 27.7–33.0 % of aseptic explants were obtained, respectively. However, after the third subculture, the yield of aseptic viable explants decreased till 10 % as a result of secondary contamination by endophytic fungi. The addition of biocide (“Gavrish”) in a nutrient medium at a concentration of 1 ml/l helped to increase the yield of aseptic viable explants till 50 %. However, after the third subculture the photosynthetic activity and the pigments content as well as growth rate decreased. Plants dropped yellowish leaves and eventually died. Thus, 37.35 % of plantlets survived after 8 months of conservation, and only 14.6 % survived after 10 months. Even after the third month of conservation significant decrease in the viability index and the coefficient of photosynthetic activity occurred in plants. Chlorophyll a in leaves decreased from 1.59 to 1.14 mg/g during 12 months in vitro conservation. The similar tendency observed on clorophyll b and carotenoids content. The experiments were carried out for 5 years using different lemon cultivars and other citrus varieties and cultivars. Thus, micropropagation and slow growth in vitro conservation of valuable lemon cultivars are still problematic and requires new technical solutions due to the low growth potential of plantlets raised from the mature buds that is consistent with the data of other researchers.