Promoter variant-dependent mRNA expression of the MEF2A in longissimus dorsi muscle in cattle

The myocyte enhancer factor 2A (MEF2A) gene encodes a member of the myocyte enhancer factor 2 (MEF2) protein family that is involved in vertebrate skeletal, cardiac, and smooth muscle development and differentiation during myogenesis. According to recent studies, MEF2 genes might be major regulators of postnatal skeletal muscle growth; thus, they are considered to be important, novel candidates for muscle development and body growth in farm animals. The aim of the present study was to search for polymorphisms in the bovine MEF2A gene and analyze their effect on the MEF2A mRNA expression level in the longissimus dorsi muscle of Polish Holstein-Fresian cattle. In total, 4094?bp of the whole coding sequence and the promoter region of MEF2A were re-sequenced in 30 animals, resulting in the detection of 6 novel variants as well as one previously reported SNP. Three linked mutations in the promoter region (-780T/G, g.-768T/G, and g.-222A/G) and only two genotypes were identified in two Polish breeds (TTA/TTA and TTA/GGG). Three SNPs in the coding region [g.1599G/A (421aa), g.1626G/A (429aa), and g.1641G/A (434aa)] appeared to be silent substitutions and segregated as two intragene haplotypes: GGG and AAA. Expression analysis showed that the mutations in the promoter region are highly associated with the MEF2A mRNA level in the longissimus dorsi muscle of bulls carrying two different genotypes. The higher MEF2A mRNA level was estimated in the muscle of bulls carrying the TTA/TTA (p<0.01) genotype as compared with those with TTA/GGG. The results obtained suggest that the nucleotide sequence mutation in MEF2A might be useful marker for body growth traits in cattle.     

Multiplexing analysis of the polyspecific intrathecal immune response in multiple sclerosis

Intrathecal synthesis of the antibodies specific to neurotrofic viruses: measles (M), rubella (R), Varicella-Zoster (Z), and/or H. simplex (H), known as "MRZH-reaction" plays important diagnostic role in multiple sclerosis (MS). Whereas the analysis of the oligoclonal IgG bands provides high sensitivity, the MRZH-reaction shows high specificity, and hence these methods complement each other. For the first time we applied multiplexing bead-based technology to simultaneously analyze cerebrospinal fluid (CSF) and serum concentrations of antibodies against these viruses, and to calculate the antibody specific indices (ASI's). The method shows reasonable precision: intra-assay, 2.9-6.7%, and inter-assay, 2.0-3.2%. The results are comparable with these obtained with other methods (ELISAs), including two runs of the certified external quality control schemes. Eighty-one percent of the MS cases (n=27) and none of the sex- and age-matched controls (n=14), except one subject with "borderline" anti-measles ASI of 1.5, showed intrathecal synthesis of IgG against at least one of the viruses discussed. The ratios of the MRZH-positive cases in the MS group were: 12/22 for M, 12/19 for R, 13/26 for Z, and 7/26 for H. We conclude that the multiplexing technology can be applied as a tool to study the intrathecal immune response in the diagnosis of MS.     

Functional heterogeneity of non-small lung adenocarcinoma cell sub-populations

The morphological and functional heterogeneity of solid tumour cells can be observed in cancer cell lines cultured in vitro. We have combined analyses of microclones developed from single cells with micropore transmigration assays to demonstrate the co-existence of cellular subsets differing in morphology and motile activity, as well as Cx43 (connexin 43) and N-cadherin expression within lung carcinoma A549 populations. 'Fibroblastoid' cells, characterized by high motility, polarized morphology and plasmalemmal localization of Cx43, displayed the strongest aptitude for transmigration through narrow obstacles. Due to high mitotic activity, they maintain the whole population but can also give rise to a sub-population of quiescent and immobile 'epithelioid' cells. Our observations indicate that phenotypic transitions between the fibroblastoid and epithelioid phenotype account for the heterogeneity of metastable A549 cell populations.     

A comparative analysis of proteins that accumulate during the initial stage of root hair development in barley root hair mutants and their parent varieties

The mechanisms of root hair formation have been studied extensively in Arabidopsis but knowledge about these processes in monocot species is still limited, especially in relation to the proteome level. The aim of this study was to identify the proteins that are involved in the initiation and the early stage of root hair tip growth in barley using two-dimensional (2D) electrophoresis and mass spectrometry. A comparison of proteins that accumulate differentially in two root hair mutants and their respective parent varieties resulted in the identification of 13 proteins that take part in several processes related to the root hair morphogenesis, such as the control of vesicular trafficking, ROS signalling and homeostasis, signal transduction by phospholipids metabolism and ATP synthesis. Among the identified proteins, two ATP synthases, two ABC transporters, a small GTPase from the SAR1 family, a PDI-like protein, a monodehydroascorbate reductase, a C2 domain-containing protein and a Wali7 domain-containing protein were found. This study is the first report on the proteins identified in the initial stage of root hair formation in barley and gives new insights into the mechanisms of root hair morphogenesis in a monocot species.     

Enterococcus in wound infections: virulence and antimicrobial resistance

Enterococci, a complex group of facultative pathogens have become increasingly isolated in various hospital settings. They are considerable frequently cultured from traumatic and surgical wounds. We investigated 57 strains of the species E. faecalis, E. faecium and E. casseliflavus isolated from infected wounds. Their ability to produce virulence factors and their sensitivity to antibiotics were evaluated using phenotypic and genotyping methods. In the phenotype studies, significant portion of the isolates produced biofilm (66.7%) and gelatinase (36.8%). Nearly 30% of the strains expressed hemolytic properties. Only a few produced DNAse (15.8%) and lipase (7.0%). The genes esp, gelE, cylA, cylB, cylM and agg were detected in most of the isolates (38.6-87.7%). All the isolated enterococci were susceptible to vancomycin and were characterized by their low resistance to antibiotics, except aminoglycosides (HLR).     

Effect of climate,topography and habitat on species-richness of breeding birds in Poland

We developed predictive maps of farmland and forest species richness which are based on the “focal species concept” and remote sensing data. We explored environmental preferences of 17 farmland and 33 forest species in Poland, using data from the Pan-European Monitoring Scheme. The largest number of farmland species was noted in the central and eastern parts, while the opposite trend was observed in the case of forest species, where the preferred areas were in the north and south-east. The most important environmental component affecting the bird faunas of farmland and forest was a gradient from arable fields to forest. This relationship is unimodal, which means that on a meso-geographical scale the highest species richness occurred in heterogeneous landscapes where fields are interspersed with forest. Our results also indicate that the geographical gradient in Poland's bird fauna can simply be attributed to the habitat-based distinction: western and central parts are dominated by large agricultural habitats and coniferous forest, while in other areas predominate mixed forests and extensively used farmland. Among the climatic factors, only rainfall influenced the farmland bird species. Its effect is non-linear, but positive, which means that in areas with higher rainfall more species are recorded.Our predictive maps are complementary to research on population trends, and can be an essential tool for effective management and conservation of species populations on a trans-national scale.     

One-pot synthesis of new (1,3-thiazolo[5,4-b]pyridin-2-yl)benzenediols and their antiproliferative activities against human cancer cell lines

A one-pot synthesis of new 4-(1,3-thiazolo[5,4-b]pyridin-2-yl)benzene-1,3-diols has been described. The compounds were prepared by the reaction of sulfinylbis[(2,4-dihydroxyphenyl)methanethione] derivatives, with various substituents in the aryl rings, with 2-chloropyridin-3-amines. Their structures were deduced from IR and, (1) H- and (13) C-NMR spectroscopic, mass spectrometric, and elemental analyses. The antiproliferative properties of some of the products against human cancer cell lines were comparable to those of cisplatin. Structure-activity analysis showed that the presence of hydrophobic substituents in both heterocyclic fused and phenyl rings of the compounds improves their biological effects. Further, an additional OH group in the resorcinol moiety reduced the antiproliferative activity.     

CYP17 and CYP19 genetic variants are not associated with age at natural menopause in Polish women

The aim of the study was to investigate associations between two common polymorphisms of CYP17 and CYP19, encoding key enzymes of estrogen biosynthesis, and age at menopause in Polish women. One hundred fifty women after menopause (49.5 ± 3.8 years), with no previous history of hormone replacement therapy took part in the study. The genetic control group consisted of 150 newborns from the same population. We investigated an association between the age at menopause and the single nucleotide polymorphism T → C in the 5′ untranslated region (promoter) of the CYP17 gene (c.-34T>C; rs743572 – MspA1) or the number of tetranucleotide repeats [TTTA]_n (rs60271534) including deletion/insertion (D/I) of a 3 bp sequence in intron 4 of the CYP19 gene. CYP17 polymorphism was analyzed by PCR-RFLP and CYP19 by PCR and capillary electrophoresis. In the case of CYP17 polymorphism, 28.7% and 36.7% wild homozygous (TT), 50.7% and 46.0% heterozygous (TC), as well as 20.6% and 17.3% mutated homozygous (CC) types were identified in the subjects and controls, respectively. The frequency of mutated alleles (C) was 46.0% vs. 40.3% (p = 0.19). In the case of CYP19 polymorphism, 34.0% and 32.0% of homozygotes (1_1), 50.7% and 51.3% of heterozygotes (1_2), 15.3% and 16.7% of homozygotes (2_2) were identified in the subjects and controls, respectively. No association between the studied CYP17 or CYP19 polymorphisms and age at menopause was found in Polish women.