Phylogenetic relationships among Porodaedalea pini from Poland and related Porodaedalea species

Porodaedalea pini has been found to be a pathogen of Pinus banksiana (1 specimen) and Pinus sylvestris (39 specimens) in north-western Poland. This fungus was initially identified by its host preferences and morphological characters of sporophores and basidiospores. The ITS 1/2 rDNA region was sequenced and analysed using the neighbor-joining, maximum likelihood and maximum parsimony methods. All P. pini from Poland, P. pini neotype and other P. pini isolates from Europe grouped together forming a moderately supported monophyletic clade. The clade included two groups which did not correlate with geographic ranges. Nucleotide polymorphism of the Polish isolates of P. pini was small. This study provides evidence for the taxonomy of some isolates of the Porodaedalea Holarctic Group in North America: grouping with P. laricis or with P. gilbertsonii suggests that the isolates belong to these species. The absence of P. pini (in a form recognized in Europe) in North America is suggested. Sequencing of the ITS 1/2 rDNA region with the basidiomycete-specific primers (ITS1-F and ITS4-B) proved to be a suitable and sufficient method for differentiation of species within the genus Porodaedalea.     

The impact of increased soil risk elements on carotenoid contents

A pot experiment was conducted to compare the responses of a non-transgenic tobacco plant (WT) and plants with genetically prolonged life-span (SAG) to risk elements of As, Cd and Zn. Plants were grown in control soil and in soil with higher levels of risk elements. The pigment contents were established by HPLC and chlorophyll fluorescence parameters were measured from slow kinetics after a 15 min dark period with the PAM fluorometer. Top (i.e. young) leaves of both WT and SAG plants were more sensitive to photoinhibition caused by these risk elements but plants showed acclimation to such elements in the bottom leaves. Plants differed in the participation of individual pigments of xanthophyll cycle: increased levels of risk elements seem to stimulate especially first (violaxanthin to antheraxanthin) and second (anhtheraxanthin to zeaxanthin) steps of the cycle in WT plants. In SAG plants, toxic elements caused an increase in the content, particularly of the initial compound of the cycle — violaxanthin.     

Biological properties of novel chitosan-based composites for medical application as bone substitute

Hydroxyapatite is the main inorganic component of bones and teeth. In order to improve mechanical properties and surgical handiness of bioceramics, a plasticizing agent e.g. polysaccharide can be added. Chitosan is a polysaccharide with biological properties that make it an ideal component of bioceramics-based composites for medical application as bone substitute. In this study, biocompatibility of two types of novel krill chitosan-based composites was evaluated. In vitro experiments were carried out using human foetal osteoblast cell line. Cytotoxicity, cell adhesion, and bone ALP activity tests were performed to assess biocompatibility of the composites. Osteoblast growth on composites was observed using confocal microscope. Our results demonstrated that fabricated novel composites are non-toxic, are favorable to cell adhesion and growth, and provoke increase in b-ALP activity with time, thus inducing osteoblast differentiation. Based on this data composites have promising clinical potential as a bone defect filler in regenerative medicine. It is worth emphasizing that our work resulted in fabrication of flexible and surgical handy, bone substitutes that possess absolute biocompatibility with structural and mechanical properties similar to trabecular bone.     

Chemical composition,antimicrobial, antioxidative and anticholinesterase activity of Satureja Montana L. ssp montana essential oil

The present study investigates the chemical compositions of three Satureja montana L. ssp montana essential oils and correlates chemical variability with biological activities. GC/MS analysis showed that with an increase in altitude (100–500–800 m), a higher content of linalool, terpinen-4-ol and cis-sabinene hydrate was found, while the percentage of phenolic compounds, thymol and carvacrol decreased. Antimicrobial activity of the essential oils was tested against 7 fungal and 23 bacterial strains. The essential oil characterized by the highest content of phenols and alcohols exhibited the highest antimicrobial potential. The correlation analysis showed that the major carriers of the obtained antioxidant activity are oxygenated monoterpenes. All essential oils inhibited human serum cholinesterase activity. High antimicrobial potential, together with moderate antioxidant capacity and strong inhibition of human serum cholinesterase, classifies S. montana essential oil as a natural source of compounds that can be used in the treatment of foodborne and neurological diseases, wound and other infections, as well as for general health improvement.     

Combined pharmacological therapy of the acute radiation disease using a cyclooxygenase-2 inhibitor and an adenosine A3 receptor agonist

Combined approaches to the treatment of acute radiation disease are preferred to single-agent therapies due to proven or anticipated better outcomes comprising increased therapeutic efficacy and decreased incidence of undesirable side effects. Our studies on post-exposure treatment of mice irradiated by sublethal or lethal doses of ionizing radiation included testing the effectiveness of meloxicam, a cyclooxygenase-2 inhibitor, and IB-MECA, an adenosine A3 receptor agonist. The efficacy of meloxicam and IB-MECA to positively influence the progress of the acute radiation disease has been tested in situations of their combined administration with granulocyte colony-stimulating factor (G-CSF) or with each other. The results of our studies revealed a significantly improved regeneration of hematopoietic cell populations ranging from the bone marrow progenitor cells to mature blood cells following combined treatments. Also, survival of mice exposed to lethal radiation doses was highest in the animals treated with a combination of the two drugs. It can be inferred from the results that if the drug combinations employed were used in humans, e.g. in the treatment of victims of radiation accidents, a better therapeutic outcome could be expected. Therefore, further studies directed at clinical applications of meloxicam and IB-MECA in radiation victims is recommended.     

Differentiated impact of procyanidins from evening primrose on human breast cancer cells

This study examines some of the biological activities of an evening primrose flavanol preparation (EPFP) against non-invasive human breast cancer cells (MCF-7). The results are compared with those obtained for highly invasive human breast cancer cells (MDAMB-231). The results show, for the first time, that EPFP reduces MCF-7 cell number, IC₅₀ = 75 µM gallic acid equivalents/GAE for 72 h incubation, and reduces migration to 52% of the control value at 100 µM L^?1 GAE. EPFP caused favorable changes in Bcl-2/Bax mRNA ratio, which rendered MCF-7 cells more sensitive to apoptosis: the number of apoptotic cells increased 2.2-fold vs. control at 100 µM GAE. Furthermore, 100 µ M L^?1 GAE EPFP caused a 1.8-fold reduction in the activity of metalloproteinase-9 (MMP-9) secreted to the culture medium by MCF-7 cells. Moreover, EPFP suppressed the expression of selected genes of matrix metalloproteinases (MMPs), a proliferation marker (Ki67), and vascular endothelial growth factor (VEGF). In conclusion, the results of this study suggest that EPFP may exhibit proapoptotic, antiproliferative, antimigratory, and antimetastatic potential towards both selected human breast cancer cell lines, which is more pronounced in the case of the highly invasive MDA-MB-231 cells.     

A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus

Influenza is a contagious disease of humans and animals caused by viruses belonging to the Orthomyxoviridae family. The influenza A virus genome consists of negative sense, single-stranded, segmented RNA. Influenza viruses are classified into subtypes based on two surface antigens known as hemagglutinin (H) and neuraminidase (N). The main problem with influenza A viruses infecting humans is drug resistance, which is caused by antigenic changes. A few antiviral drugs are available, but the most popular is the neuraminidase inhibitor — oseltamivir. The resistance against this drug has probably developed through antigenic drift by a point mutation in one amino acid at position 275 (H275Y). In order to prevent a possible influenza pandemic it is necessary to develop fast diagnostic tests. The aim of this project was to develop a new test for detection of influenza A virus and determination of oseltamivir resistance/sensitivity in humans. Detection and differentiation of oseltamivir resistance/sensitivity of influenza A virus was based on real-time PCR. This test contains two TaqMan probes, which work at different wavelengths. Application of techniques like multiplex real-time PCR has greatly enhanced the capability for surveillance and characterization of influenza viruses. After its potential validation, this test can be used for diagnosis before treatment.     

In vitro organogenesis secondary metabolite production and heavy metal analysis in Swertia chirayita

An efficient protocol of plant regeneration through direct and indirect organogenesis in Swertia chirayita was developed. Explants cultured on Murashige and Skoog medium supplemented with 2,4-D (0.5 mg L^?1) with combination of Kinetin (0.5 mg L^?1) showed the highest frequency (84%) of callusing and 1.0mg L^?1 6-benzyladenine (BA) in combination with (100 mg L^?1) Adenine sulphate (Ads) + (0.1 mg L^?1) Indole acetic acid (IAA) was excellent for maximum adventitious shoot (12.69 ± 1.30) formation in four week of culture. A maximum number of (7.14 ± 0.99) shoots were developed per leaf explants through direct organogenesis. The highest frequency of rooting (11.46 ± 1.56) was observed on MS medium augmented with IAA (1.0 mg L^?1). Well-rooted shoots transferred to plastic pots containing a soilrite: sand mix and then moved to the greenhouse for further growth and development. Four major secondary metabolites were analyzed and quantified using high performance liquid chromatography. Amount of secondary metabolites was found significantly higher, in in vitro plantlets compared to in vivo plantlets and callus raised from S. chirayita. Higher heavy metal accumulation in in vitro as compared to in vivo plantlets correlates higher secondary metabolite production supporting that they play regulatory role in influencing the plant secondary metabolism.     

Head and Neck Squamous Cell Carcinoma: Prognosis using molecular approach

Head and neck squamous cell carcinoma (HNSCC) is the fifth most prevalent cancer worldwide. Apart from various known clinicopathogical factors, it is still a major concern as many genetic and epigenetic alterations bring about the possibility of this deadly disease. The aim of this review is to explore the possible role of DNA repair pathways and the polymorphic status of DNA repair genes (XPA, XPC, XPD, XRCC1 and XRCC3) in the onset of HNSCC, along with sequence variations in genes such as Glutathione S-transferases (GSTT1, M1 and P1) that are significantly associated with HNSCC risk. We also focus on the p53 gene mutation induced by various etiological agents and threat factors with its implications towards HNSCC, and emphasise the current therapeutic interventions in treating HNSCC.     

The action of CGRP and SP on cultured skin fibroblasts

Background/purpose

Calcitonin gene-related peptide (CGRP) is the most abundant neuropeptide in the skin, followed by substance P (SP), vasoactive intestinal peptide (VIP), and other neuropeptides in smaller amounts. The proliferative effect of neuropeptides on fibroblasts may affect wound healing and may be associated with hyperproliferative skin and mesenchymal disorders. Understanding the neuropeptidergic action on fibroblasts may provide relevant information to a deeper comprehension of the healing process. This study reviews the action of the main neuropeptides, CGRP and SP, on cultured human skin fibroblasts.

Methods

A systematic literature search was conducted on Medline and Web of Science databases on December 21, 2013.

Results

A total of 74 articles were retrieved using the proposed search strategies and 3 were found in the references section of the selected articles. Thirteen of the retrieved articles studied the action of CGRP and SP on cultured human skin fibroblasts, 12 of which related to SP and 1 related to both CGRP and SP.

Conclusion

Only one study was retrieved about the action of both CGRP and SP on cultured human skin fibroblasts. Further studies are necessary to investigate CGRP on skin fibroblasts and its role in the fibroplasia phase of wound healing.     

Identification of proteins involved in starch and polygalacturonic acid degradation using LC/MS

Plant biomass in the form of cheap wastes, such as straw, corn stalks, wood chips, sawdust, bagasse, pomace, etc., is abundant throughout the world. To convert these wastes into the useful value-added compounds microbial enzymes are the preferred choice. In this paper, we identify enzymes involved in the degradation of starch and polygalacturonic acid using liquid chromatography/mass spectrometry based analysis. We analysed total protein from soil and compost samples. Extracellular proteins from enrichment cultures were analysed in parallel and used as controls in the sample preparation and identification of proteins. In general, both protein sequence coverage and the number of identified peptides were higher in the samples obtained from the enrichment cultures than from the total protein from soil and compost. The influence of the nature of gel (zymography vs. SDS/polyacrylamide) was negligible. Thus, starch and polygalacturonic acid degradation associated proteins can be directly excised from the zymograms without the need to align zymograms with the SDS/polyacrylamide gels. A range of starch and polygalacturonic acid degradation associated enzymes were identified in both total protein samples and extracellular proteins from the enrichment cultures. Our results show that proteins involved in starch and polygalacturonic acid degradation can be identified by liquid chromatography/mass spectrometry from the complex protein mixtures both with and without cultivation of microorganism     

Erratum to: “The Luc2 gene enhances reliability of bicistronic assays”

Paper by Masek et al. “The Luc2 gene enhances reliability of bicistronic assays” in Volume 8, Issue 5, 423–431 / May 2013; DOI: 10.2478/s11535-013-0151-z contains incomplete graphic file inserted as Figure 2. The correct Figure 2, together with its caption is presented below.     

Erratum to Central European Journal of Biology, Volume 8, Issue 9

Paper by Osyczka and Rola “Cladonia lichens as the most effective and essential pioneers in strongly contaminated slag dumps” in Volume 8, Issue 9, 876–887 / September 2013; DOI: 10.2478/s11535-013-0210-0 contains incorrect units in Table 2 and Table 3. The correct Tables 2 and 3, together with their captions are presented below.     

Occurrence of Diploid Lines of <Emphasis Type="Italic">Puccinia graminis tritici</Emphasis> in Axenic Culture

THE dikaryophase of the wheat stem rust fungus, Puccinia graminis (Pers.) f. sp. tritici(Erikss. and E. Henn.), can now be grown on artificial medium^1,2; but cultures of this organism initiated from heavy inocula of urediospores usually sporulate or stale after 2-4 weeks and the growth period cannot be appreciably prolonged by subculture onto fresh medium^2,3. Scott and Maclean³, however, referred to the discovery of continuously subculturable isolates of P. graminis tritici. These were obtained by incubating urediospores on liquid nutrient medium, but Scott and Maclean offered no information which might explain the distinctive behaviour of the isolates. We have obtained lines of P. graminis tritici from race 126-Anz 6,7 (culture 70165) which will grow continuously on artificial medium. This communication describes how these lines were obtained and presents evidence that their mycelia are composed of diploid cells.     

Erratum to Central European Journal of Biology, Volume 8, Issue 12

Paper by Bogdan Jaroszewicz, Ewa Piro?nikow, Marcin Churski “Vegetation diversity influences endozoochoric seed dispersal by moose (Alces alces L.)“ in Voulme 8, Issue 12, 1250-1264/ December 2013; DOI: 10.2478/s11535-013-0244-3 contains incorrect affiliations of Ewa Piro?nikow and Marcin Churski. Ewa Pi?niow works in Institute of Biology, University of Bia?ystok, 15-950 Bia?ystok, Poland; while Marcin Churski works in Mammal Research Institute, Polish Academy of Sciences, 17–230 Bia?owie?a, Poland.     

The Phylogenetic Distribution and Evolution of Enzymes Within the Thymidine Kinase 2-like Gene Family in Metazoa

Deoxyribonucleoside kinases (dNKs) carry out the rate-determining step in the nucleoside salvage pathway within all domains of life where the pathway is present, and, hence, are an indication on whether or not a species/genus retains the ability to salvage deoxyribonucleosides. Here, a phylogenetic tree is constructed for the thymidine kinase 2-like dNK gene family in metazoa. Each enzyme class (deoxycytidine, deoxyguanosine, and deoxythymidine kinases, as well as the multisubstrate dNKs) falls into a monophyletic clade. However, in vertebrates, dCK contains an apparent duplication with one paralog lost in mammals, and a number of crustacean genomes (like Caligus rogercresseyi and Lepeophtheirus salmonis) unexpectedly contain not only the multisubstrate dNKs, related to Drosophila multisubstrate dNK, but also a TK2-like kinase. Additionally, crustaceans (Daphnia, Caligus, and Lepeophtheirus) and some insects (Tribolium, Danaus, Pediculus, and Acyrthosiphon) contain several multisubstrate dNK-like enzymes which group paraphyletically within the arthropod clade. This might suggest that the multisubstrate dNKs underwent multiple rounds of duplications with differential retention of duplicate copies between insect families and more complete retention within some crustaceans and insects. Genomes of several basal animalia contain more than one dNK-like sequence, some of which group outside the remaining eukaryotes (both plants and animals) and/or with bacterial dNKs. Within the vertebrates, the mammalian genomes do not contain the second dCK, while birds, fish, and amphibians do retain it. Phasianidae (chicken and turkey) have lost dGK, while it has been retained in other bird lineages, like zebra finch. Reconstruction of the ancestral sequence between the multisubstrate arthropod dNKs and the TK2 clade of vertebrates followed by homology modeling and discrete molecular dynamics calculations on this sequence were performed to examine the evolutionary path which led to the two different enzyme classes. The structural models showed that the carboxyl terminus of the ancestral sequence is more helical than dNK, in common with TK2, although any implications of this for enzyme specificity will require biochemical validation. Finally, rate-shift and conservation-shift analysis between clades with different specificities uncovered candidate residues outside the active site pocket which may have contributed to differentiation in substrate specificity between enzyme clades.     

Isolation and characterization of a lead (Pb) tolerant <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> strain HF5 for decolorization of reactive red-120 and other azo dyes

Presence of heavy metals including lead (Pb) in the textile effluents is a crucial factor affecting the growth and potential of the dye decolorizing bacterial strains. This work was planned to isolate and characterize a bacterial strain exhibiting the potential to decolorize a range of azo dyes as well as the resistance to Pb. In this study, several Pb tolerant bacteria were isolated from effluents of textile industry. These bacterial isolates were screened for their potential of decolorizing the reactive red-120 (RR120) azo dye with presence of Pb (50 mg L^?1). The most efficient isolate was further characterized for its potential to resist Pb and decolorize different azo dyes under varying cultural and incubation conditions. Out of the total 82 tested bacterial isolates, 30 bacteria were found to have varying potentials to resist the presence of lead (Pb) and carry out decolorization of an azo dye reactive red-120 (RR120) in the medium amended with Pb (50 mg L^?1). The most efficient selected bacterium, Pseudomonas aeruginosa strain HF5, was found to show a good potential not only to grow in the presence of considerable concentration of Pb but also to decolorize RR120 and other azo dyes in the media amended with Pb. The strain HF5 completely (>?90%) decolorized RR120 in mineral salt medium amended with 100 mg L^?1 of Pb and 20 g L^?1 NaCl. This strain also considerably (>?50%) decolorized RR120 up to the presence of 2000 mg L^?1 of Pb and 50 g L^?1 of NaCl but with reduced rate. The optimal decolorization of RR120 by HF5 was achieved when the pH of the Pb amended (100 mg L^?1) mineral salt media was adjusted at 7.5 and 8.5. Interestingly, this strain also showed the tolerance to a range of metal ions with varying MIC values. The Pseudomonas aeruginosa strain HF5 harboring the unique potentials to grow and decolorize the azo dyes in the presence of Pb is envisaged as a potential bioresource for devising the remediation strategies for treatment of colored textile wastewaters loaded with Pb and other heavy metal ions.     

The Evolution of Reproduction-Related NLRP Genes

NLRP proteins are important components of inflammasomes with a major role in innate immunity. A subset of NLRP genes, with unknown functions, are expressed in oocytes and early embryos. Mutations of Nlrp5 in mice are associated with maternal-effect embryonic lethality and mutations of NLRP7 in women are associated with conception of biparental complete hydatidiform moles (biCHMs), suggesting perturbed processes of genomic imprinting. Recessive mutations on NLRP2/7 in humans are associated with reproductive disorders and appear to be induced by a demethylation of the maternal pronucleus. In this study, we find that radiation of NLRP genes occurred before the common ancestor of Afrotheria and Boreoeutheria, with the clade of oocyte-expressed genes originating before the divergence of marsupial and eutherian mammals. There have been multiple independent duplications of NLRP2 genes one of which produced the NLRP7 gene associated with biCHMs.     

Erratum to: Central European Journal of Biology, Volume 6, Issue 5

Paper by Péli et al. “Ecophysiological responses of desiccation-tolerant cryptobiotic crusts” in Volume 6, Issue 5, 838–849 / October 2011; DOI: 10.2478/s11535-011-0049-1 contains incorrect graphic file inserted as Figure 6. The correct Figure 6, together with its caption is presented below.