Band 7, Heft 2

Protein kinase CK2 is a highly conserved Ser/Thr protein kinase involved in cell cycle control, transcription, signal transduction and cell proliferation. It is upregulated in several diseases and by oxidative stress. CK2 is generally composed of two catalytic subunits and two regulatory subunits and utilizes either ATP or GTP as a phosphate donor. CK2 was isolated from the sea mussel Mytilus galloprovincialis, a biomarker of marine pollution, and the Mediterranean fly Ceratitis capitata, an insect capable of wreaking extensive damage to a wide range of fruit crops with great economical importance. The catalytic CK2α and regulatory CK2β subunits of M. galloprovincialis and C. capitata show similar properties. The mussel and fly catalytic subunits and holoenzymes were capable of phosphorylating the recombinant ribosomal stalk P1 protein, implying functional conservation. They also demonstrate the characteristics of a typical CK2: use of ATP and GTP as phosphate donors, inhibition by known modulators of CK2 activity (like benzotriazole derivatives and heparin), and stimulation by polycations. Both organisms seem to be ideal models for the analysis of CK2 in the control of gene expression in response to cellular stress.

It is generally accepted that the location of body fat deposits may play an important role in the risk of developing some endocrine and metabolic diseases. We have studied the effect of food restriction and food restriction/refeeding, often practiced by individuals trying to lose body weight, on the expression of genes which are associated with obesity and certain metabolic disorders in inguinal, epididymal, and perirenal rat white adipose tissues. Gene expression was analyzed by real time semi-quantitative polymerase chain reaction and by Western blot. We found that prolonged food restriction caused a significant decrease of body and adipose tissue mass as well as the increase of Scd1 and Elovl6 gene expressions in all main rat adipose tissue deposits. Food restriction/refeeding caused increases of: a) Scd1 and Elovl6 mRNA levels in adipose tissue, b) Scd1 protein level and c) desaturation index in adipose tissue. The increased expression of both genes was unusually high in inguinal adipose tissue. The results suggest that the increase of Scd1 and Elovl6 gene expressions in white adipose tissue by prolonged food restriction and prolonged food restriction/refeeding may contribute to accelerated fat recovery that often occurs in individuals after food restriction/refeeding.

A considerable variability in the incidence and prevalence of diabetic nephropathy (DN) coheres with an important contribution of multigenetic predisposition in the development of DN. Some genes, which probably participate in the pathogenesis of diabetic nephropathy, also play a role in the regulation of blood pressure, familial hyperlipidemia, familial hypertension and other diseases of the cardiovascular system. We have examined the association of diabetic nephropathy, nephropathy of non-diabetic origin, hypertension and of type 2 diabetes itself with several genetic polymorphisms (the insertion/deletion polymorphism in the gene for angiotensin-converting enzyme, the G/T polymorphism in the glucose transporter 1 gene, the G/T (894) polymorphism and the T/C (−786) polymorphism in the eNOS gene in three groups of patients with diabetes mellitus: 1) patients without diabetic nephropathy (DM); 2) patients with DN; 3) patients with nephropathy of non-diabetic origin (NDRD). Angiotensin-converting enzyme is an important factor in a development of arterial hypertension, but in our groups of Central European diabetic patients the I/D polymorphism was not associated with diabetic nephropathy. Furthermore, we have confirmed that the T/C (T786C) polymorphism in the eNOS gene is associated with metabolic syndrome including type 2 diabetes.

E. coli biofilms cause serious problems in medical practice by contaminating surfaces and indwelling catheters. Due to the rapid development of antibiotic resistance, alternative approaches to biofilm suppression are needed. This study addresses whether products released by antagonistic bacteria — Lactobacillus isolates from vaginal and dairy-product samples could be useful for controlling E. coli biofilms. The effects of diluted cell-free supernatants (CFS) from late-exponential Lactobacillus cultures on the growth and biofilm production of Escherichia coli were tested. Most of the CFS applied as 10−2 had no impact on bacterial growth, biofilm development however was influenced even by 10−4 of CFS. Initial screening by crystal violet assay showed that biofilm modulation varied between different CFS and E. coli combinations from inhibition to activation; however three of the tested CFS showed consistency in biofilm suppression. This was not due to antibacterial activity since Live/Dead fluorescence labeling showed insignificant differences in the amount of dead cells in control and treated samples. Some E. coli strain-specific mechanisms of response to the three CFS included reduction in hydrophobicity and motility. Released exoploysaccharides isolated from the three CFS stimulated sessile growth, but proteinase K reduced their inhibitory activities implying participation of protein or peptide biofilm suppression factor(s).

Optimal detection of pathogens by molecular methods in water samples depends on the ability to extract DNA rapidly and efficiently. In this study, an innovative method was developed using a microfluidic biochip, produced by microelectrochemical system technology, and capable of performing online cell lysis and DNA extraction during a continuous flow process. On-chip cell lysis based on chemical/physical methods was performed by employing a sufficient blend of water with the lysing buffer. The efficiency of lysis with microfluidic biochip was compared with thermal lysis in Eppendorf tubes and with two commercial DNA extraction kits: Power Water DNA isolation kit and ForensicGEM Saliva isolation kit in parallel tests. Two lysing buffers containing 1% Triton X-100 or 5% Chelex were assessed for their lysis effectiveness on a microfluidic biochip. SYBR Green real-time PCR analysis revealed that cell lysis on a microfluidic biochip using 5% Chelex buffer provided better or comparable recovery of DNA than commercial isolation kits. The system yielded better results for Gram-positive bacteria than for Gram-negative bacteria and spores of Gram-positive bacteria, within the limits of detection at 103 CFU/ml. During the continuous flow process in the system, rapid cells lysis with PCR-amplifiable genomic DNA were achieved within 20 minutes.

Members of the yeast genus Malassezia, including atypical strains, are lipophilic except for Malassezia pachydermatis. New physiological features that characterize atypical Malassezia strains are mainly associated with alteration in Tween assimilation pattern — such isolates still require lipids for growth. We isolated three non-lipid-dependent strains of Malassezia from patients with diagnosed atopic dermatitis (AD). These isolates could not be identified to the species level via their physiological properties. Phylogenetic trees, based on the D1/D2 regions of the 26S rDNA gene sequences and nucleotide sequences of the ITS1-5.8S-ITS2 rRNA region, showed the isolates to belong to Malassezia furfur. Three non-lipid dependent isolates from AD skin were conspecific, and sequences analysis proved them to be M. furfur.

The paper discusses the dynamics of the accumulation of microscopic fungi, depending on the sludge load (Bx), in activated sludge used for landfill leachate pretreatment. The propagule washout from the sludge into pretreated leachates is determined, including genera and species that may threaten environmental health. An increased accumulation of microscopic fungi in sludge flocs occurred at Bx=0.23−0.45 mg chemical oxygen demand (COD) mg−1 d−1. Microscopic fungi were eluted at the maximal Bx value tested of 1.64 mg COD mg−1 d−1. Both the activated sludge and the leachate runoff from the sequencing batch reactor (SBR) pose health risks to the environment due to the occurrence of fungi such as Aspergillus fumigatus, Purpureocillium lilacinum, Cyberlindnera jadinii (C. utilis), Geotrichum candidum and G. fragrans. Their count is sufficient to cause multi-organ infections in homeothermal animals and in humans.

Keratinophilic fungi are a highly specialized, keratin-degrading ecological group. They live in natural environments, mostly in the keratinrich remains of dead animal in the soil. We investigated species diversity in four types of soils with different physico-chemical properties. The strain material was identified based on morphological characters. Different representatives of Chrysosporium and geophilic dermatophytes dominated depending on soil pH. Geophilic dermatophytes were represented by one species, Trichophyton ajelloi, and the Chrysosporium group was represented by Chrysosporium keratinophilum. The frequency of Trichophyton ajelloi increased with an increase in pH, and it reached the maximum in strongly acidic soil (podzol), unlike the Chrysosporium group. The frequency of Chrysosporium keratinophilum was positively correlated with the content of humus, nitrogen, CaCO3 and phosphorus in the soils.

The chestnut blight pathogen Cryphonectria parasitica was studied in a chestnut collection composed of both seedlings and grafts derived from selected Castanea sativa and C. sativa × C. crenata trees located in south-east Slovakia, near village Príbelce on an area of approximately 3.5 ha. The study was conducted during eight years (2003–2010). During this period 133 trees were infected, which represents 59.82% of chestnut trees of all chestnut accessions. Based on the phenotype of the fungus culture and the type of cankers in the field, all isolates were determined to be virulent. No hypovirulent strains were found. No vegetative compatibility (vc) type diversity was observed. More than 130 isolates were analyzed for vc and all were in single vc type, which was identical with EU 12. All isolates assayed for mating type were MAT-1. No perithecia were observed. No significant differences were found between the proportion of cankered and dead cankered trees in seedlings and grafts of hybrid origin (C. sativa × C. crenata) and of C. sativa origin. However, particular seedlings and grafts of hybrid origin seemed to exhibit certain resistance to chestnut blight.

In the present study, we have evaluated the effects of increased UV-B radiation that simulates 17% ozone depletion, on fungal colonisation and concentrations of rutin, catechin and quercetin in common buckwheat (Fagopyrum esculentum) and tartary buckwheat (Fagopyrum tataricum). Induced root growth and reduced shoot:root ratios were seen in both of these buckwheat species after enhanced UV-B radiation. There was specific induction of shoot quercetin concentrations in UV-B-treated common buckwheat, whereas there were no specific responses for flavonoid metabolism in tartary buckwheat. Root colonisation with arbuscular mycorrhizal fungi significantly reduced catechin concentrations in common buckwheat roots, and induced rutin concentrations in tartary buckwheat, but did not affect shoot concentrations of the measured phenolics. Specific UV-B-related reductions in the density of microsclerotia were observed in tartary buckwheat, indicating a mechanism that potentially affects fungus-plant interactions. The data support the hypothesis that responses to enhanced UV-B radiation can be influenced by the plant pre-adaptation properties and related changes in flavonoid metabolism.

The paper presents data on the remarkable abundance of S. graffii in the parks of the industrially transformed region of Upper Silesia, Poland. Ten new sites and 36 colonies were recorded, while only 6 sites were known so far from the whole territory of Poland. New host plant species and new ant species tending aphids were also recorded. The conclusion is that the abundance of Stomaphis species may be underestimated due to their cryptic life mode.

During 2009–2011, mosquitoes were captured in CDC miniature light traps using CO2 (dry ice) at six sites in the Lower Dyje River Basin (Czech Republic). Other methods of capture — sweeping from vegetation and collection of larvae and pupae from ponds — were also used for more precise diagnostics. Thirty mosquito species of six genera were confirmed. A total of 415,218 females were captured. Most frequently found were the outbreak species Aedes vexans (56.52% of all mosquitoes collected) and Ae. sticticus (16.40%). Among other flood species, Ae. rossicus (5.17%), Ae. cantans and Ae. annulipes (2.44% of all females collected), and Ae. cinereus s. l. (1.11%) were especially abundant. Females of Ae. cataphylla were captured in spring (0.31%) and Ae. intrudens was numerous only at one site. Among the other species, Culex pipiens s. l. (6.61%) and Cx. modestus (8.87%) were abundant. Anopheles maculipennis s. l. (1.01%), An. claviger (0.43%), An. plumbeus (0.08%), An. hyrcanus (0.08%), Coquillettidia richiardii (0.52%) and Culiseta annulata (0.18%) were also detected. Sparsely occurring were Ae. excrucians, Ae. flavescens, Ae. caspius and Ae. geniculatus. Captured only very sporadically were Ae. communis, Ae. leucomelas, Ae. dorsalis, Ae. rusticus, Cx. martinii, Cx. territans, Cs. morsitans and Uranotaenia unguiculata.

Oxidative stress is accepted to play a significant role in stress symptoms, caused by different stressors in a variety of organisms. In this study seedlings of spring barley (Hordeum vulgare L.) were exposed to a wide range of copper, zinc, chromium, nickel, lead and cadmium concentrations in order to determine the relationships between heavy metals-induced oxidative stress and plant growth inhibition. All investigated heavy metals induced an essential increase in lipid peroxidation and a reduction of dry biomass along with an increase in metal concentration in the nutrient solution. A very close and statistically significant exponential relationship between lipid peroxidation and growth inhibition was detected in this study. According to the results of analysis of variance (ANOVA), the intensity of nonspecific oxidative stress is identified as the main factor of barley growth inhibition, explaining 75% of total variance. Almost 10% of growth inhibition is attributed to the specific impact of heavy metals. The most pronounced increase of malondialdehyde content and growth inhibition was observed in Cu and Cd treatments, whereas the lowest changes in observed indicators were detected after exposure to Zn and Pb.

We investigated the concentration of Aluminium (Al), Cobalt (Co), Chromium (Cr), Copper (Cu), Iron (Fe), Manganese (Mn), Nickel (Ni) and Zinc (Zn) in the root and aboveground organs of four halophyte species (Salicornia europaea, Suaeda maritima, Salsola soda and Halimione portulacoides), as well as in the soil from maritime and inland saline areas. The aim of our research was to evaluate the capability of some halophyte species to absorb different heavy metals and to detect differentiation of heavy metal accumulation within populations from inland and maritime saline areas. Generally, the plant roots had significantly higher concentrations of metals when compared to stems and leaves. Zinc was the only metal with concentrations significantly higher in the leaves than in the root and stem. Populations from maritime saline areas had higher trace root and stem metal concentrations than populations from inland saline areas. Excepting zinc, populations from inland saline areas had higher heavy metal concentrations in the leaves. The factors that affected metal accumulation by halophytes included the percentage of salt in the soil. We also discuss the potential use of these halophytes in phytoremediation.

We presented a methodology for drawing continuous boundaries in the landscape differentiating between regions with different floristic composition. A region in Central Slovakia covering 2,445 km2 was investigated. Ecological indicator values for temperature (EIT) in 1,978 grassland polygons were analysed. Ordinary kriging was used to interpolate EIT across the study region. Lattice wombling was used to identify the most intensive gradients in EIT and to draw boundaries, while ANOVA was used for post-classification analysis. A strong pattern of spatial continuity was present in EIT assigned to species in grassland polygons allowing for drawing continuous boundaries in the landscape. The study region was divided into 15 districts using the proposed method. Post-classification analysis indicated that 17 out of 23 adjacent districts were found to differ significantly in term of mean value of source samples. The results implied the need for incorporating spatial autocorrelation in sample data into post-classification analysis; such factor is often neglected in ecological research. The presented findings suggested broader applicability of the proposed method for spatial modelling, as vegetation data is widely accessible in databases for many regions of Europe.

The objective of this study was to evaluate the efficacy of the ethanolic extract of endemic plant Halacsya sendtneri in inhibiting the growing of the test fungi and bacteria as well as to determine its genotoxic potential and toxicity using the Allium anaphase-telophase assay. Minimum inhibitory concentrations (MIC) were determined for 15 indicator strains of pathogens, representing both bacteria and fungi. The highest susceptibility to the ethanolic extract of H.sendtneri was exhibited by Pseudomonas glycinea (FSB4), (MIC=0.09 mg/ml) among the bacteria, and by Phialophora fastigiata (FSB81), (MIC=1.95 mg/ml) among the fungi. The composition of H. sendtneri extracts was also determined using HPLC analysis. Rosmarinic acid was found to be the dominant phenolic compound. The Allium anaphase-telophase genotoxicity assay revealed that the ethanolic extract of H. sendtneri at concentrations of 31.5 mg/l and below does not produce toxic or genotoxic effects. This is the first report of chemical constituents, genotoxic and antimicrobial activities of the endemic species, H. sendtneri.

Vitamin E has generated immense interest because of its potential of being an antioxidant, a neuroprotector, and a protector against atherosclerosis, carcinogenesis and cardiovascular disease. However, the prooxidant chemistry of vitamin E cannot be ignored since it is related to the generation of peroxyl radicals. In the present study, 125, 250 and 500 mg/kg of vitamin E-acetate (VE) administered intraperitoneally (i.p.) to Balb/C mice significantly induced 6%, 8% and 11.33% (control value=2.33%) of chromosome aberrations (CA) and 0.88%, 1.39% and 1.81% (control value=0.61%) of micronucleus (MN), following 24 hour of treatment in the bone marrow cells. In the germ cells, VE did not induce any sperm head abnormality (SHA) after 35 days of exposure. Most importantly, it has been observed that pre-treatment with VE significantly reduces CA, MN, and SHA induction by chemotherapeutic drug cisplatin (CIS). Our findings suggest that lone treatment with VE induce genotoxicity in somatic cells after 24 and 48 hours of exposure but not in germ cells after 35 days of exposure, whereas pre-treatment with VE reduces CIS induced genotoxicity as well as cytotoxicity. There exists a thin line of difference on the behavioral transition of VE when acting alone and when acting with a drug.

Lung cancer continues to represent a major public health concern with high morbidity and mortality worldwide. Early detection of lung cancer is problematic due to a lack of diagnostic markers with high sensitivity and specificity. To determine the differently expressed proteins in the serum of lung cancer and identify the function of such proteins, two-dimensional electrophoresis (2DE) and liquid chromatography mass spectrometry (LC-MS) were used to screen the serum of lung cancer model induced by 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK). A total of 25 protein spots were qualitatively different and 6 were quantitatively different in the serum from rats bearing induced lung cancer when compared with normal controls. Two of the proteins that showed major changes in concentration in sera were identified to be Immunoglobulin γ 2A chain C region (heavy chain) and Transferrin by LC-MS/MS.

Lectins are important components of the immune defense system of invertebrates. Given their important functions, numerous investigations have been carried out on the characterization and function of lectins in invertebrates. However, lectin studies with the freshwater planarian, an evolutionarily important animal, are rare. In this paper, we demonstrate agglutination of glutaraldehyde treated erythrocytes by a lectin with preference for rabbit erythrocytes. The result of hemagglutinating activity inhibition assays with several carbohydrates showed the most potent inhibitor was maltose. A natural lectin from the crude homogenates of freshwater planarian Dugesia japonica was purified by single step affinity chromatography using amylose-coupled agarose. The purified protein appeared as one band with a molecular mass of 350 kDa in PAGE, and as one band, approximately 56 kDa, in SDS-PAGE. The purified lectin showed dependence on calcium. The activity of the purified lectin was inhibited at temperatures greater than 50°C and showed a pH optimum between 5–8. The purified lectin also has binding activity to the Gram-negative bacteria E. coli, and the Gram-positive bacteria B. subtilis. Furthermore, the purified lectin obtained from injured and bacteria-induced planarians showed increased agglutinating activity against rabbit erythrocytes. These results suggest that the purified lectin may play an important role in the innate immunity of the freshwater planarian.

Paper by Hussein et al. “Effectiveness of the entomopathogenic nematode Steinernema feltiae in agar gel formulations against larvae of the Colarado potato beetle, Leptinotarsa decemlineata (Say.) (Coleoptera: Chrysomelidae)” in Volume 7, Issue 1, 77–82 9 / February 2012; DOI: 10.2478/s11535-011-0090-0 contains an error in the title. The correct title is presented below.