bluuebunny/crossref_metadata_2025 huggingface dataset:
Nucleotide substitutions in the Candida albicans ERG11 gene of azole-susceptible and azole-resistant clinical isolates.
Authors: Strzelczyk Joanna Katarzyna , Slemp-Migiel Anna , Rother Magdalena , Gołąbek Karolina , Wiczkowski Andrzej
Published: February 2019

One of the mechanisms of Candida albicans resistance to azole drugs used in antifungal therapy relies on increased expression and presence of point mutations in the ERG11 gene that encodes sterol 14α demethylase (14DM), an enzyme which is the primary target for the azole class of antifungals. The aim of the study was to analyze nucleotide substitutions in the Candida albicans ERG11 gene of azole-susceptible and azole-resistant clinical isolates. The Candida albicans isolates represented a collection of 122 strains selected from 658 strains isolated from different biological materials. Samples were obtained from hospitalized patients. Fluconazole susceptibility was tested in vitro using a microdilution assay. Candida albicans strains used in this study consisted of two groups: 61 of the isolates were susceptible to azoles and the 61 were resistant to azoles. Four overlapping regions of the ERG11 gene of the isolates of Candida albicans strains were amplified and sequenced. The MSSCP (multitemperature single strand conformation polymorphism) method was performed to select Candida albicans samples presenting genetic differences in the ERG11 gene fragments for subsequent sequence analysis. Based on the sequencing results we managed to detect 19 substitutions of nucleotides in the ERG11 gene fragments. Sequencing revealed 4 different alterations: T495A, A530C, G622A and A945C leading to changes in the corresponding amino acid sequence: D116E, K128T, V159I and E266D. The single nucleotide changes in the ERG11 gene did not affect the sensitivity of Candida albicans strains, whereas multiple nucleotide substitutions in the ERG11 gene fragments indicated a possible relation with the increase in resistance to azole drugs.
A computational approach to structural properties of glycoside hydrolase family 4 from bacteria.
Authors: Craciun Dana , Vlad-Oros Beatrice , Filimon Nicoleta , Ostafe Vasile , Isvoran Adriana
Published: February 2019

Structural bioinformatics approaches applied to the alpha- and beta-glycosidases from the GH4 enzyme family reveal that, despite low sequence identity, these enzymes possess quite similar global structural characteristics reflecting a common reaction mechanism. Locally, there are a few distinctive structural characteristics of GH4 alpha- and beta-glycosidases, namely, surface cavities with different geometric characteristics and two regions with highly dissimilar structural organizations and distinct physicochemical properties in the alpha- and beta-glucosidases from Thermotoga maritima. We suggest that these structurally dissimilar regions may be involved in specific protein-protein interactions and this hypothesis is sustained by the predicted distinct functional partners of the investigated proteins. Also, we predict that alpha- and beta-glycosidases from the GH4 enzyme family interact with difenoconazole, a fungicide, but there are different features of these interactions especially concerning the identified structurally distinct regions of the investigated proteins.
Dynamics of reactive oxygen species generation in the presence of copper(II)-histidine complex and cysteine.
Authors: Ząbek-Adamska Anna , Drożdż Ryszard , Naskalski Jerzy W
Published: February 2019

Histidine-copper(II) complex (Cu-His2) is a form of bound copper necessary for cellular copper uptake. Due to the high affinity of histidine to copper(II) ions, the binding of copper(II) by histidine is considered a substantial part of plasma antioxidative defense. Also cysteine plays a role in the antioxidative system. However, we show here that in the presence of oxygen the histidine-copper(II) complex plus cysteine produces reactive oxygen species (ROS). Cysteine concentration was assayed using a thiol specific silver-mercury electrode. Hydrogen peroxide was assayed amperometrically using platinum electrode. ROS formation was followed by chemiluminescence of luminol-fluoresceine-enhanced system. Addition of cysteine to Cu-His2 solution at pH 7.4 in the presence of atmospheric oxygen initiates the synthesis of H2O2 and generation of ROS, which manifests as a burst of chemiluminescence. The reaction has two stages; in the first stage, cysteine is utilized for the synthesis of an unstable intermediary product which becomes a substrate for ROS formation. Anaerobic conditions inhibit ROS formation. Increased cysteine concentration enhances the lag phase of the oxidative burst without influencing the amount of ROS. The synthesis of ROS (measured by chemiluminescence) is proportional to the concentration of Cu-His2 employed. ROS production can be repetitively initiated by further additions of cysteine to the reaction medium. The study suggests that Cu-His2 catalyzes cysteine-dependent reduction of oxygen to superoxide employing an intermediary cysteine-copper(I) complex and enabling Fenton reaction with copper and hydrogen peroxide produced as a secondary product. In effect, Cu-His2 with cysteine may be a source of ROS in biological media.
Evaluation of the activity of thermostable DNA polymerases in the presence of heme, as a key inhibitor in the real time PCR method in diagnostics of sepsis.
Authors: Gosiewski Tomasz , Brzychczy-Włoch Monika , Pietrzyk Agata , Sroka Agnieszka , Bulanda Małgorzata
Published: February 2019

The study aim was evaluation of the usefulness of several thermostable DNA polymerases in real time PCR conducted in the presence of the heme. Our study had the advantage of testing several different polymerases, one of which proved to be the least sensitive to heme activity. We also found that there is no need of supplementing the reaction mixture with protective substances like BSA. Selection of the appropriate polymerase can increase the efficiency of the PCR reaction which is very important for diagnosis of sepsis and for other analyses performed on DNA template isolated from the blood.
Molecular genetics of PKU in Poland and potential impact of mutations on BH4 responsiveness.
Authors: Bik-Multanowski Miroslaw , Kaluzny Lukasz , Mozrzymas Renata , Oltarzewski Mariusz , Starostecka Ewa , Lange Agata , Didycz Bozena , Gizewska Maria , Ulewicz-Filipowicz Jolanta , Chrobot Agnieszka , Mikoluc Bozena , Szymczakiewicz-Multanowska Agnieszka , Cichy Wojciech , Pietrzyk Jacek J
Published: February 2019

Tetrahydrobiopterin (BH4) has been recently approved as a treatment of patients with phenylketonuria. However, as a confirmation of BH4-responsiveness, it might require a very expensive trial treatment with BH4 or prolonged BH4-loading procedures. The selection of patients eligible for BH4-therapy by means of genotyping of the PAH gene mutations may be recommended as a complementary approach. A population-wide genotyping study was carried out in 1286 Polish phenyloketonuria-patients. The aim was to estimate the BH4 demand and to cover prospectively the treatment by a National Health Fund. A total of 95 types of mutations were identified. Genetic variants corresponding with probable BH4-responsiveness were found in 28.2% of cases. However, patients with mild or classical phenylketonuria who require continuous treatment accounted for 11.4% of the studied population only. Analysis of the published data shows similar percentage of the "BH4-responsive" variants of a PAH gene in patients from other countries of Eastern Europe. Therefore, it can be concluded, that the proportion of phenylketonuria-patients who could benefit from the use of BH4 reaches approximately 10% in the entire region.
Generation of stable, non-aggregating Saccharomyces cerevisiae wild isolates.
Authors: Wloch-Salamon Dominika M , Plech Marcin , Majewska Jagoda
Published: February 2019

Cellular aggregates observed during growth of Saccharomyces cerevisiae strains derived from various natural environments makes most laboratory techniques optimized for non-aggregating laboratory strains inappropriate. We describe a method to reduce the size and percentage of the aggregates. This is achieved by replacing the native allele of the AMN1 gene with an allele found in the W303 laboratory strain. The reduction in aggregates is consistent across various environments and generations, with no change in maximum population density or strain viability, and only minor changes in maximum growth rate and colony morphology.
Prolonged renal failure in the course of atypical ethylene glycol intoxication.
Authors: Liberek Tomasz , Śliwarska Julita , Czurak Krzysztof , Perkowska-Ptasińska Agnieszka , Weber Ewa , Rutkowski Bolesław
Published: February 2019

Ethylene glycol poisoning is not an uncommon cause of an acute renal injury. In this paper we present case of prolonged renal failure in the course of ethylene glycol intoxication. Due to the low dose of ingested ethylene glycol and concomitant ethanol consumption, the neurodepressive phase of the intoxication was rather mild and patient presented to the hospital on the ninth day after poisoning with established renal failure. The diagnosis of the specific cause of renal injury was facilitated by the renal biopsy.
The use of infrared spectroscopy and artificial neural networks for detection of uropathogenic Escherichia coli strains' susceptibility to cephalothin.
Authors: Lechowicz Lukasz , Urbaniak Mariusz , Adamus-Białek Wioletta , Kaca Wiesław
Published: February 2019

Infrared spectroscopy is an increasingly common method for bacterial strains' testing. For the analysis of bacterial IR spectra, advanced mathematical methods such as artificial neural networks must be used. The combination of these two methods has been used previously to analyze taxonomic affiliation of bacteria. The aim of this study was the classification of Escherichia coli strains in terms of susceptibility/resistance to cephalothin on the basis of their infrared spectra. The infrared spectra of 109 uropathogenic E. coli strains were measured. These data are used for classification of E. coli strains by using designed artificial neural networks.
The most efficient artificial neural networks classify the E. coli sensitive/resistant strains with an error of 5%.
Bacteria can be classified in terms of their antibiotic susceptibility by using infrared spectroscopy and artificial neural networks.
Antibiotics sensitivity of Candida clinical and food-borne isolates.
Authors: Maroszyńska Marta , Kunicka-Styczyńska Alina , Rajkowska Katarzyna , Maroszyńska Iwona
Published: February 2019

Candida is a yeast species recognized as the most frequent etiological agent of systemic and invasive thrush in humans. Invasions can affect all tissues, organs and systems of human in various stages of development. In the last 10 years Candida infections have increased 15 times. The purpose of our study was to determine the sensitivity of four antibiotics belonging to three different groups of antifungal agents against clinical and food-borne Candida strains. Our studies showed that of all tested strains, 7% was resistant to nystatin, 32% to fluconazole, 23% to voriconazole, and no strains grew in the presence of caspofungin. Despite the differences in biochemical profiles of clinical and food-borne isolates of Candida, a group of strains showing resistance to antibiotics include both types of isolates. At the same time circulating of antibiotic-resistant strains outside the hospital environment and the yeast infection via food is possible.
Production of triterpenoids with cell and tissue cultures.
Authors: Malinowska Magdalena , Sikora Elżbieta , Ogonowski Jan
Published: February 2019

Triterpenes are group of biologically active compounds which can be found in higher plants. Their main source are plants bark, leaves, twigs, fruits, resins or oils. The biological activity of triterpens is very diversified and many studies have already confirmed the following therapeutic effects: anti-inflammatory, antimicrobial, antiviral, antifungial, immunomodulatory, and hepatoprotective. Synthesis of triterpenes derivates can be performed by chemical or enzymatic reactions, however biotransformation is more specific and eliminates the side products and the molecule alterations. These processes use isolated enzymes or microorganisms. Cell culture in vitro eliminates problems like extract variability as well as instability of the compounds being obtained during the extraction process. What is more, it ensures high reproducibility and optimal regio- and enantioselectivity. The most widely used technique is a classical screening of a series of microbial strains. Studies on triterpene biotransformation give a lot of information about new biologically active compounds and let predict the metabolism of biological compounds. This review presents most important advancements in the metabolic engineering of microorganisms for the production of triterpenoids. Moreover, the review highlights general strategies to obtain rich biochemical diversity of plants by employing the biocatalysts produced by microorganisms or tissue cultures.
Intra-population genetic diversity of cultivated carrot (Daucus carota L.) assessed by analysis of microsatellite markers.
Authors: Maksylewicz Anna , Baranski Rafal
Published: February 2019

Intra-population variation of 18 cultivated carrot (Daucus carota L. ssp. sativus) populations of diverse origins was evaluated using codominant microsatellite (SSR) markers. Using 27 genomic and EST-derived SSR markers, 253 alleles were identified with a mean 9.4 alleles per marker. Most of the alleles (60.5%) were rare i.e., with the frequency ≤ 0.05 while only 3.95% of alleles occurred with frequency > 0.6. EST-derived SSR markers were less polymorphic than genomic SSR markers. Differences in allele occurrence allowed 16 out of 18 populations to be assigned to either the Western or Asian carrot gene pools with high probability. Populations could be also discriminated due to the presence of private alleles (25.3% of all alleles). Most populations had excess of alleles in the homozygous state indicating their inbreeding, although heterozygous loci were common in F1 hybrids. Genetic diversity was due to allelic variation among plants within populations (62% of total variation) and between populations (38%). Accessions originating from continental Asia and Europe had more allelic variants and higher diversity than those from Japan and USA. Also, allelic richness and variability in landraces was higher than in F1 hybrids and open-pollinated cultivars.
Influence of process parameters on properties of Nanostructured Lipid Carriers (NLC) formulation.
Authors: Lasoń Elwira , Sikora Elżbieta , Ogonowski Jan
Published: February 2019

Nanostructured lipid carriers (NLC) are stable colloidal formulations with notable advantages for drug delivery systems. Thanks to their physicochemical stability, biocompatibility, biodegradability and controlled drug release, they have received increasing attention for the last several years. The aim of the study was to prepare and characterize nanostructured lipid carriers (NLC). Both, the effect of the process parameters and the effect of the preemulsion composition on the NLC properties were investigated. In the work, different type of surfactants (i.e. decyl glucoside, Poloxamer188, Tween 80, sodium cholate) and their combinations were used to stabilize NLC dispersions. Moreover, several kinds of solid lipids (modified beeswax, gliceryl behenate, cetyl palmitate and berry wax) and liquid lipids (caprilic/capric triglyceride and decyl oleate) were applied. An ultrasonication method using a probe type sonicator was used to obtain NLC, and the time and energy of the process were modified throuhout. The physicochemical properties of the formulations, such as particle size, size distribution, polidispersity index were studied using the dynamic light scattering (DLS) method. The electrophoretic mobility of obtained particles was also measured, using the Zetasizer Nano ZS Malvern Instrument based on the Laser Doppler Velocimetry (LDV) technique. Knowing the value of electrophoretic mobility of particles for given conditions, the zeta potential was determined. The obtained results showed that the process parameters and the composition of the preemulsion had significant impact on the nanoparticles structure. The optimal formulations size ranged between 60 and 80 nm, and the value of their zeta potential was up to -30mV. The stability of these systems was further confirmed by macroscopic observation.
Studies on the formation of O/W nano-emulsions, by low-energy emulsification method, suitable for cosmeceutical applications.
Authors: Jaworska Małgorzata , Sikora Elżbieta , Zielina Michał , Ogonowski Jan
Published: February 2019

The formation of oil/water (O/W) nano-emulsions suitable for cosmeceutical application was studied. Nano-emulsions were prepared by using phase inversion composition (PIC) method, one of the low-energy emulsification methods. The process consist of stepwise water addition to oil/surfactant mixture, at T = 25°C. Caprylic/capric triglycerides (GTCC), propylene glycol dicaprylate/dicaprate (PC) and oleic acid (OA) were applied as an oil phase. Polysorbate 80 was used as the surfactant. Kinetic stability of the nano-emulsions was analyzed by measuring droplet size as a function of time for different oil/surfactant ratio. The particles size distribution was analyzed by means DLS measurement technique (Dynamic Light Scattering), using Zetasizer Nano ZS (Malvern Instruments, UK). One of triterpenoic acid, practically non-water soluble substance was selected as an active and incorporated into the stable formulation. The obtained results proved that the nanoemulsion NE-T80-GTCC-20:80 based on caprylic/capric triglycerides with the oil/surfactant ratio O/S = 2 0:80 and the droplet size r = 25 nm was the most stable one and additionally showed the highest solubilisation capacity for the triterpene.
Changes in the properties of pig manure slurry.
Authors: Kowalski Zygmunt , Makara Agnieszka , Fijorek Kamil
Published: February 2019

The paper presents the results of analyses of samples of manure from a pig farm located near Piła, Poland performed between June 2011 and May 2012 using a single sampling system. The statistical analyses of the average content of chemical and biological oxide demands, nitrogen, phosphorus, potassium, calcium and dry mass in the slurry in various seasons allowed us to draw conclusions concerning the changes in the chemical composition of the manure in specific seasons and to determine the correlations between the chemical parameters. The average content of N, BOD, P, and dry mass content tended to decrease systematically from the spring until the winter. The highest correlation coefficient, which indicates significant interdependency among the variables tested, was consistently found for COD and BOD, whereas the smallest correlation coefficient was found consistently for K and Ca and once for Ca and N.
Arbutin production via biotransformation of hydroquinone in in vitro cultures of Aronia melanocarpa (Michx.) Elliott.
Authors: Kwiecień Inga , Szopa Agnieszka , Madej Kornelia , Ekiert Halina
Published: February 2019

Arbutin (hydroquinone β-D-glucoside) is a compound of plant origin possessing valuable therapeutic (urinary tract disinfection) and cosmetic (skin whitening) properties, which can be obtained from in vitro cultures of plants belonging to different taxa via biotransformation of exogenously supplemented hydroquinone. Agitating cultures of Aronia melanocarpa were maintained on the Murashige and Skoog medium containing growth regulators: the cytokinin - BAP (6-benzylaminopurine), 2 mg/l and the auxin NAA (α-naphthaleneacetic acid), 2 mg/l. The biomass was cultured for 2 weeks and then hydroquinone was supplemented at the following doses: 96, 144, 192, 288 and 384 mg/l either undivided or divided into two or three portions added at 24-hour intervals. The content of the reaction product - arbutin, was determined using an HPLC method in methanolic extracts from biomass and lyophilized medium samples collected 24 hours after the addition of the last precursor dose. The total amounts of arbutin were very diverse, from 2.71 to 8.27 g/100g d.w. The production of arbutin rose with increasing hydroquinone concentration. The maximum content of the product was observed after hydroquinone addition at 384 mg/l divided into two portions. Biotransformation efficiency also varied widely, ranging from 37.04% do 73.80%. The identity of the product - arbutin, after its isolation and purification was confirmed by spectral analysis ((1)H-NMR spectrum). The maximum amount of arbutin obtained was higher than that required by the latest 9(th) Edition of the Polish Pharmacopoeia and by the newest 8th Edithion of European Pharmacopoeia for Uvae ursi folium (7.0 g/100g d.w.), and is interesting from practical point of view.
Purinergic signaling in the pancreas and the therapeutic potential of ecto-nucleotidases in diabetes.
Authors: Cieślak Marek , Roszek Katarzyna
Published: February 2019

It is widely accepted that purinergic signaling is involved in the regulation of functions of all known tissues and organs. Extracellular purines activate two classes of receptors, P1-adenosine receptors and P2-nucleotide receptors, in a concentration-dependent manner. Ecto-enzymes metabolizing nucleotides outside the cell are involved in the termination of the nucleotide signaling pathway through the release of ligands from their receptors. The pancreas is a central organ in nutrient and energy homeostasis with endocrine, exocrine and immunoreactive functions. The disturbances in cellular metabolism in diabetes mellitus lead also to changes in concentrations of intra- and extracellular nucleotides. Purinergic receptors P1 and P2 are present on the pancreatic islet cells as well as on hepatocytes, adipocytes, pancreatic blood vessels and nerves. The ATP-dependent P2X receptor activation on pancreatic β-cells results in a positive autocrine signal and subsequent insulin secretion. Ecto-NTPDases play the key role in regulation of extracellular ATP concentration. These enzymes, in cooperation with 5'-nucleotidase can significantly increase ecto-adenosine concentration. It has been demonstrated that adenosine, through activation of P1 receptors present on adipocytes and pancreatic islets cells, inhibits the release of insulin. Even though we know for 50 years about the regulatory role of nucleotides in the secretion of insulin, an integrated understanding of the involvement of purinergic signaling in pancreas function is still required. This comprehensive review presents our current knowledge about purinergic signaling in physiology and pathology of the pancreas as well as its potential therapeutic relevance in diabetes.
Role of mitochondria in carcinogenesis.
Authors: Tokarz Paulina , Blasiak Janusz
Published: February 2019

Mitochondria play the central role in supplying cells with ATP and are also the major source of reactive oxygen species (ROS) - molecules of both regulatory and destructive nature. Dysfunction of mitochondrial metabolism and/or morphology have been frequently reported in human cancers. This dysfunction can be associated with mitochondrial DNA (mtDNA) damage, which may be changed into mutations in mtDNA coding sequences, or the displacement-loop region, changes in the mtDNA copy number or mtDNA microsatellite instability. All these features are frequently associated with human cancers. Mutations in mtDNA can disturb the functioning of the ROS-producing organelle and further affect the entire cell which may contribute to genomic instability typical for cancer cells. Although the association between some mtDNA mutations and cancer is well established, the causative relationship between these two features is largely unknown. A hint suggesting the driving role of mtDNA mutations in carcinogenesis comes from the observation of tumor promotion after mtDNA depletion. Mitochondria with damaged DNA may alter signaling of the mitochondrial apoptosis pathway promoting cancer cell survival and conferring resistance to anticancer drugs. This resistance may be underlined by mtDNA copy number depletion. Therefore, mitochondria are considered a promising target in anticancer therapy and several mitochondria-targeting drugs are in preclinical and clinical trials. Some other aspects of mitochondrial structure and functions, including morphology and redox potential, can also be associated with cancer transformation and constitute new anticancer targets. Recently, several studies have disclosed new mechanisms underlying the association between mitochondria and cancer, including the protection of mtDNA by telomerase, suggesting new approaches in mitochondria-oriented anti-cancer therapy.
Elevated high sensitivity C-reactive protein and uric acid levels in coronary artery ectasia.
Authors: Demir Şerafettin , Karakoyun Gulhan , Kanadasi Mehmet
Published: February 2019

The aim of this study is to examine uric asid (UA) and high sensitive C-Reactive protein (Hs-CRP) levels in patients with coronary artery ectasia (CAE).
Ninety-eight patients with isolated CAE (mean age 57.5±10.3), (group-I), 110 patients with CAD but without CAE (mean age 56.3±10.7), (group-II), and 105 patients with normal coronary angiographies (mean age 58.1±10.8), (group-III) were included in the study. Blood samples of all individual were taken after coronary angiography from an antecubital vein, the patients uric acid and Hs-CRP levels were assessed. The severity of ectasia was evaluated and categorized according to Markis.
A significant difference was not seen in serum uric acid and Hs-CRP levels between CAE and CAD groups. However, relative to the control group, uric acid and Hs-CRP levels in CAE and CAD groups were higher to a significant degree (p=0.001, p
Discrete dynamic system oriented on the formation of prebiotic dipeptides from Rode's experiment.
Authors: Polanco Carlos , Samaniego José Lino , Buhse Thomas , Castañón González Jorge Alberto
Published: February 2019

This work attempts to rationalize the possible prebiotic profile of the first dipeptides of about 4 billion years ago based on a computational discrete dynamic system that uses the final yields of the dipeptides obtained in Rode's experiments of salt-induced peptide formation (Rode et al., 1999, Peptides 20: 773-786). The system built a prebiotic scenario that allowed us to observe that (i) the primordial peptide generation was strongly affected by the abundances of the amino acid monomers, (ii) small variations in the concentration of the monomers have almost no effect on the final distribution pattern of the dipeptides and (iii) the most plausible chemical reaction of prebiotic peptide bond formation can be linked to Rode's hypothesis of a salt-induced scenario. The results of our computational simulations were related to former simulations of the Miller, and Fox & Harada experiments on amino acid monomer and oligomer generation, respectively, offering additional information to our approach.
Differentiation by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) of Candida albicans isolated from upper respiratory tract in patients with non-small cell lung cancer.
Authors: Biernasiuk Anna , Korona-Głowniak Izabela , Grzegorczyk Agnieszka , Malm Anna
Published: February 2019

Cancer patients are predisposed to fungal infections caused by Candida albicans, especially to oral or respiratory tract candidiasis. The aim of this study was to estimate genetic diversity by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) of C. albicans isolated from upper respiratory tract of 100 patients with non-small cell lung cancer. Among 52 strains, 34 genotypes were defined. 10 clusters comprising 28 (53.85%) isolates with similarity coefficient ≥ 80% were formed. The remaining 24 (46.15%) isolates represented individual genotypes. The RAPD-PCR technique revealed genomic variability within C. albicans isolated from upper respiratory tract of the cancer patients.
Cystain C and neuropeptid Y levels in brain tissues after experimental subarachnoid hemorrhage.
Authors: Açıkgöz Şerefden , Can Murat , Güven Berrak , Edebali Nurullah , Barut Figen , Büyükuysal Çağatay , Tekin İshak Özel , Açıkgöz Bektaş
Published: February 2019

The aim of this study was to investigate the changes in the levels of cystatin C, which protects neurodegeneration in the central nervous system with the inhibition of cysteine protease and by inducing autophagy in the pathogenesis of cerebral vasospasm and levels of vasoconstrictive neuropeptid Y (NPY) in the brain tissue homogenates of rat model of subarachnoid hemorrhage (SAH). Three experimental groups were used: Day 2 and Day 7 groups after SAH, and also a control group. There were seven Wistar albino rats in each group. SAH was accomplished by transclival basilar artery puncture. Rat cystatin C, rat NPY were determined with ELISA in brain tissue homogenates. Day 2 group showed significantly enhanced cystatin C values in comparision with the control group (P=0.048). NPY levels between the Day 2 and Day 7 groups and the control groups were not significantly different (P=0.315). In histopathological examination, there was less neuronal loss in the Day 2 group than in the Day 7 group. Regarding our results, it would be more valuable to measure NPY levels in specific brain areas. The increased cystatin C levels on the second day after SAH is probably a pathophysiologic mechanism to organize protease activity.
In vivo reassortment of influenza viruses.
Authors: Urbaniak Kinga , Markowska-Daniel Iwona
Published: February 2019

The genetic material of influenza A virus consists of eight negative-sense RNA segments. Under suitable conditions, the segmented structure of the viral genome allows an exchange of the individual gene segments between different strains, causing formation of new reassorted viruses. For reassortment to occur, co-infection with two or more influenza virus strains is necessary. The reassortment is an important evolutionary mechanism which can result in antigenic shifts that modify host range, pathology, and transmission of the influenza A viruses. In this process, the influenza virus strain with epidemic and/or pandemic potential can be created. Cases of this kind were in 1957 (Asian flu), 1968 (Hong Kong flu) and recently in 2009 (Mexico). Viruses containing genes of avian, swine, and/or human origin are widespread around the world, for example the triple reassortant H1N1 virus causing the 2009 influenza pandemic in 2009 that has become a seasonal virus. The aim of the study is to present the mechanism of reassortment and the results of experimental co-infection with different influenza viruses.
Avian influenza in Poland.
Authors: Smietanka Krzysztof , Minta Zenon
Published: February 2019

Poland has experienced four episodes of avian influenza (AI) outbreaks over the past two decades. The first epidemic was caused by a low pathogenicity (LPAIV) H7N7 subtype and occurred in fattening and breeder turkeys in 1995. Two waves of H5N1 high pathogenicity avian influenza (HPAI) took place in 2006 and 2007. In spring 2006, 64 cases of the H5N1 virus were detected, mostly in mute swans. In December 2007, ten outbreaks of H5N1 HPAI were detected in commercial poultry (n = 9) and wild birds kept in captivity (n = 1). The outbreaks in 2006 and 2007 were caused by genetically similar but clearly distinguishable viruses of the 2.2 clade. In 2013, an H9N2 avian influenza virus was detected in 4 fattening turkey holdings. The virus was low pathogenic and a phylogenetic study has shown a close relatedness to the Eurasian lineage of AIV of the wild bird origin. Neither preventive nor prophylactic vaccinations have ever been used in poultry or other birds. Emergency vaccinations using autogenous vaccine were introduced only to control the H7N7 LPAI outbreaks in 1995. The baseline surveillance for AI in live migratory birds and poultry provides a valuable insight into the ecology of AIV at the wild and domestic bird interface. Passive surveillance is in place of early detection of HPAIV infection in dead or moribund birds.
Detection of the influenza virus yesterday and now.
Authors: Woźniak-Kosek Agnieszka , Kempińska-Mirosławska Bogumiła , Hoser Grażyna
Published: February 2019

Demographic changes and the development of transportation contribute to the rapid spread of influenza. Before an idea of a 'person to person' spread appeared, divergent theories were developed to explain influenza epidemics in the past. Intensified virological and serological tests became possible after isolation of the human influenza virus in 1933. The first influenza virus detection methods were based on its isolation in egg embryos or cell lines and on demonstration of the presence of the viral antigens. Molecular biology techniques associated with amplification of RNA improved the quality of tests as well as sensitivity of influenza virus detection in clinical samples. It became possible to detect mixed infections caused by influenza types A and B and to identify the strain of the virus. Development of reliable diagnostic methods enabled fast diagnosis of influenza which is important for choosing an appropriate medical treatment.
Virus-like particles as vaccine.
Authors: Chroboczek Jadwiga , Szurgot Inga , Szolajska Ewa
Published: February 2019

This review presents data on commercial and experimental virus-like particle (VLP) vaccines, including description of VLP vaccines against influenza. Virus-like particles are multimeric, sometimes multiprotein nanostructures assembled from viral structural proteins and are devoid of any genetic material. VLPs present repetitive high-density displays of viral surface proteins. Importantly, they contain functional viral proteins responsible for cell penetration by the virus, ensuring efficient cell entry and thus tissue-specific targeting, determined by the origin of the virus. The foremost application of VLPs is in vaccinology, where they provide delivery systems that combine good safety profiles with strong immunogenicity and constitute a safe alternative to inactivated infectious viruses. These stable and versatile nanoparticles display excellent adjuvant properties capable of inducing innate and cognate immune responses. They present both, high-density B-cell epitopes, for antibody production and intracellular T-cell epitopes, thus inducing, respectively, potent humoral and cellular immune responses. Uptake of VLPs by antigen-presenting cells leads to efficient immune responses resulting in control of pathogenic microorganisms.
Expression of avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) under control of the ptcB promoter in Lactococcus lactis.
Authors: Szatraj Katarzyna , Szczepankowska Agnieszka K , Sączyńska Violetta , Florys Katarzyna , Gromadzka Beata , Łepek Krzysztof , Płucienniczak Grażyna , Szewczyk Bogusław , Zagórski-Ostoja Włodzimierz , Bardowski Jacek
Published: February 2019

Gram-positive and nonpathogenic lactic acid bacteria (LAB) are considered to be promising candidates for the development of new, safe systems of heterologous protein expression. Recombinant LAB has been shown to induce specific local and systemic immune response against selected pathogens, and could be a good alternative to classical attenuated carriers. The main goal of our study was to express the avian influenza haemagglutinin (H5) and chicken interleukin 2 (chIL-2) in Lactococcus lactis. Results of this study were anticipated to lead to construction of lactococcal strain(s) with potential vaccine properties against the avian influenza A (H5N1) virus. Expression of the cloned H5 gene, its His-tagged variant and chIL-2 gene, under the control of the ptcB gene promoter was attested by RT-PCR on transcriptional level and Western or dot blot analysis on translational level, demonstrating that system can be an attractive solution for production of heterologous proteins. The results of the preliminary animal trial conducted in mice are a promising step toward development of a vaccine against avian bird flu using Lactococcus lactis cells as antigen carriers.
Pilot study on feasibility of application of gas chromatography for the assessment of acrylamide concentration in sewage sludge.
Authors: Włodarczyk Elżbieta , Próba Marta , Wolny Lidia , Wojtal Łukasz
Published: February 2019

The aim of this study was to determine the possibility of using gas chromatography to measurement of the acrylamide concentration in sewage sludge. Acrylamide, as a toxic substance, is not indifferent to human health, but it is used in the production of plastics, dyes, adhesives, cosmetics, mortar, as well as a coagulant for water treatment, wastewater or sewage sludge conditioning. Determination of acrylamide by gas chromatography was based on standard: EPA Method 8032A "Acrylamid by gas chromatography." It consists of a bromination reaction of the compound in the presence of dibromopropendial derivative, a triple extraction with the ethyl acetate, a concentration of the eluate sample up to the 1 ml volume, and an analysis by the gas chromatography using an electron capture detector (ECD). The acrylamide concentration of was calculated according to the formula presented in the mentioned standard. All samples were performed twice (the difference between the results was not greater than 10%), and the average value of the four samples was 17.64 µg/L(-1). The presence of acrylamide in sewage sludge has been confirmed.
Triterpene sapogenins with oleanene skeleton: chemotypes and biological activities.
Authors: Jatczak Kamil , Grynkiewicz Grzegorz
Published: February 2019

Critical survey of a selected class of pentacyclic triterpenes--the oleanane family, is presented based on current literature in order to underline their value for medicinal chemistry and drug development potential. Oleanenes may be considered as a renewable resource of valuable research materials which are structurally diverse, inherently biocompatible and have built-in affinity for many categories of functional proteins. Although availability of particular compounds from natural sources may be very low, synthetic methods elaborated by generations of chemists, secure a way to obtaining desirable structures from commercial starting materials.
Computational model of abiogenic amino acid condensation to obtain a polar amino acid profile.
Authors: Polanco Carlos , Buhse Thomas , Samaniego José Lino , Castañón González Jorge Alberto , Arias Estrada Miguel
Published: February 2019

In accordance with the second law of thermodynamics, the Universe as a whole tends to higher entropy. However, the sequence of far-from-equilibrium events that led to the emergence of life on Earth could have imposed order and complexity during the course of chemical reactions in the so-called primordial soup of life. Hence, we may expect to find characteristic profiles or biases in the prebiotic product mixtures, as for instance among the first amino acids. Seeking to shed light on this hypothesis, we have designed a high performance computer program that simulates the spontaneous formation of the amino acid monomers in closed environments. The program was designed in reference to a prebiotic scenario proposed by Sydney W. Fox. The amino acid abundances and their polarities as the two principal biases were also taken into consideration. We regarded the computational model as exhaustive since 200,000 amino acid dimers were formed by simulation, subsequently expressed in a vector and compared with the corresponding amino acid dimers that were experimentally obtained by Fox. We found a very high similarity between the experimental results and our simulations.
Simple molecular diagnostic method for fragile X syndrome in Egyptian patients: pilot study.
Authors: Meguid Nagwa A , Ismail Manal F , El-Mahdy Rasha S , Barakat Maged A , El-Awady Mostafa K
Published: February 2019

Poor knowledge about Fragile X syndrome (FXS) may be a major barrier to early diagnosis that could improve quality of life and prognosis especially in the developing countries.
The aim of this study was to evaluate simple and reproducible method for premutation detection in females of fragile X families for the first time in Egypt.
We have developed a rapid modified polymerase chain reaction (PCR)-based screening tool for expanded Fragile X mental retardation 1 (FMR1) alleles. This method utilizes betaine as additive to facilitate FMR 1 gene amplification. We screened fifty three males, thirty two first-degree females; twenty normal healthy controls in addition to six reference samples.
Simple PCR method showed 16 males with abnormal CGG repeats, where 10 of their mothers and four sisters had FMR 1 premutation. Consanguineous marriage was present in 66.6% percent of the studied families. Studying the correlation between genotype and clinical manifestations showed premature ovarian failure in 40% and learning disability in 50% of the studied female carriers.
FXS has to be ruled out in families with consanguineous parents, before assuming that familial mental retardation is due to autosomal recessive gene defects. Early carrier detection may reduce the number of affected children. In conclusion, more studies are still needed of much larger sample size with known allele sizes in order to guarantee the accuracy of the method used.
N-acetyl-beta-glucosaminidase urine activity as a marker of early proximal tubule damage and a predictor of the long-term function of the transplanted kidneys.
Authors: Kwiatkowska Ewa , Domański Leszek , Bober Joanna , Kłoda Karolina , Safranow Krzysztof , Szymańska-Pasternak Jolanta , Romanowski Maciej , Sulecka Aneta , Pawlik Andrzej , Ciechanowski Kazimierz
Published: February 2019

Ischaemia-reperfusion injury (IRI) is a factor leading to the damages of the transplanted kidney, what affects mainly the proximal tubules. Early monitoring of tubule damage can be an efficient tool to predict the allograft dysfunction. Present in proximal tubules, N-acetyl-beta-glucosaminidase (NAG) is a lysosomal enzyme whose excretion rises as a result of IRI or acute rejection. The aim of this study was to monitor the NAG urine activity to evaluate the early proximal tubule damage, and to try to predict the long-term function of the transplanted kidney.
The study enrolled 87 Caucasian renal transplant recipients (61.7% males, 38.3% females, mean age 45.56±14.34 years). Urine samples were collected for NAG and creatinine analysis on the 1st day after transplantation, and then in the 3rd and 12th month. Protocol biopsies were performed in the 3rd and 12th month.
A significant positive correlation between NAG urine activity in the 3rd month after transplantation and creatinine concentration on the 14th (p=0.004) and 30th day (p=0.05), in the 3rd month (p=0.009) and after the 1st (p=0.005) and 2nd year (p=0.003) was observed. A statistically significantly higher urinary NAG activity in samples collected in the first 3 days and in the 3rd month after transplantation among patients with DGF (p=0.006 and p=0.03 respectively) was found. There was a significant positive correlation between NAG urine activity in the 3rd month and the grade of tubular atrophy in specimens collected in the 3rd (p=0.03) and 12th (p=0.04) month.
Monitoring of NAG urine activity is useful in the evaluation of early proximal tubule damage and predicting the long-term function of the transplanted kidneys.
PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains.
Authors: Klesiewicz Karolina , Nowak Paweł , Karczewska Elżbieta , Skiba Iwona , Wojtas-Bonior Izabela , Sito Edward , Budak Alicja
Published: February 2019

The occurrence of clarithromycin resistance among Helicobacter pylori strains is a major cause of the treatment failure. Resistance to this drug is conferred by point mutations in 23S rRNA gene and the most prevalent mutations are A2143G and A2142G. The aim of the study was to evaluate the occurrence of A2143G and A2142G mutations in a group of H. pylori strains resistant to clarithromycin.
The study included 21 clarithromycin-resistant H. pylori strains collected between 2006 and 2009 in southern Poland. Resistance to clarithromycin was quantitatively tested with the E-test to determine the minimal inhibitory concentration (MIC value). The point mutations of H. pylori isolates were detected by PCR followed by RFLP analysis.
The MIC values for clarithromycin for the analyzed strains ranged from 1.5 mg/L to 64 mg/L. Nine H. pylori strains exhibited A2143G mutation and A2142G mutation was found in 9 isolates as well. The results of RFLP analysis of 3 clarithromycin-resistant strains were negative for both mutations. The average MIC values for A2143G and A2142G mutants were 6 and 30 mg/L, respectively.
Frequencies of A2143G and A2142G mutations were the same in all isolates tested. Strains with A2143G mutation exhibited lower MIC values than A2142G mutants. Application of PCR-RFLP method for detection of clarithromycin resistance allows for better and more efficient management of H. pylori infections.
Smooth muscle contamination analysis in clinical oncology gene expression research.
Authors: Markowska Monika , Stępniak Piotr , Wojdan Konrad , Świrski Konrad
Published: February 2019

Gene expression profiling is one of the most explored methods for studying cancers and microarray data repositories have become a rich and important resource. The most common human cancers develop in organs that are walled by smooth muscles. The only method of sample extraction free of unintentional contamination with surrounding tissue is microdissection. Nevertheless, such an approach is implemented infrequently. In the light of the above, there is a possibility of smooth muscle contamination in a large portion of publicly available data. In this study, 2292 publicly available microarrays were analysed to develop a simple screening method for detecting smooth muscle contamination. Microarray Inspector software was used to perform the tests since it has the unique ability to use many selected genes and probesets in a single group as a tissue definition. Furthermore, the test was dataset-independent. Two strategies of tissue definition were explored and compared. The first one depended on Tissue Specific Genes Database (TiSGeD) and BioGPS web resources, which themselves were based on meta-analysis of thousands of microarrays. The second method was based on a differential gene expression analysis of a few hundred preselected arrays. The comparison of the two methods proved the latter to be superior. Among the tested samples of undefined contamination, nearly half were identified to possibly contain significant smooth muscle traces. The obtained results equip researches with a simple method of examining microarray data for smooth muscle contamination. The presented work serves as an example of how to create definitions when searching for other possible contaminations.