Polymeric biomaterials with antibacterial effects are requisite materials in the fight against hospital-acquired infections. An effective way for constructing a second generation of antibacterials is to exploit the synergic effect of (i) patterning of polymeric materials by a laser, and (ii) deposition of noble metals in their nanostructured forms. With this approach, we prepared highly-ordered periodic structures (ripples) on polyethylene naphthalate (PEN). Subsequent deposition of Ag under the glancing angle of 70° resulted in the formation of self-organized, fully separated Ag nanowire (Ag NW) arrays homogenously distributed on PEN surface. Surface properties of these samples were characterized by AFM and XPS. Vacuum evaporation of Ag at the glancing angle geometry of 70° caused that Ag NWs were formed predominantly from one side of the ripples, near to the top of the ridges. The release of Ag(+) ions into physiological solution was studied by ICP-MS. The results of antibacterial tests predetermine these novel structures as promising materials able to fight against a broad spectrum of microorganisms, however, their observed cytotoxicity warns about their applications in the contact with living tissues.

• MeSH
• Anti-Bacterial Agents chemistry   pharmacology   MeSH
• Escherichia coli drug effects   MeSH
• Photoelectron Spectroscopy MeSH
• Lasers * MeSH
• Microscopy, Atomic Force MeSH
• Nanowires chemistry   toxicity   MeSH
• Polyethylene chemistry   MeSH
• Surface Properties MeSH
• Staphylococcus epidermidis drug effects   MeSH
• Silver chemistry   MeSH
• Publication type
• Journal Article MeSH

Antiretroviral restriction factors may play an essential role in the safety of xenotransplantation. Therefore, the present study focused on investigation of the changes in the tripartite motif-containing family (TRIM) gene expression in normal human dermal fibroblasts with and without lipopolysaccharide stimulation in response to porcine endogenous retrovirus infection. Analysis of the expression profile of TRIMs was performed using oligonucleotide microarrays and QRT-PCR. Nine (TRIM1, TRIM2, TRIM5, TRIM14, TRIM16, TRIM18, TRIM22, TRIM27 and TRIM31) statistically significantly differentially expressed genes were found (P < 0.05, one-way ANOVA). In conclusion, comprehensive analysis of retroviral restriction factor gene expression in human dermal fibroblasts before and after porcine endogenous retrovirus infection with and without LPS stimulation may suggest association of the selected TRIMs with antiretroviral activity.

• MeSH
• Amino Acid Motifs MeSH
• Endogenous Retroviruses physiology   MeSH
• Fibroblasts metabolism   virology   MeSH
• Humans MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Reverse Transcriptase Polymerase Chain Reaction MeSH
• Reproducibility of Results MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Dermis pathology   MeSH
• Gene Expression Profiling * MeSH
• Sus scrofa MeSH
• Carrier Proteins genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria.

• MeSH
• Down-Regulation MeSH
• Fibroblasts cytology   metabolism   microbiology   MeSH
• Cells, Cultured MeSH
• Chickens genetics   metabolism   MeSH
• Chick Embryo MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Macrophages metabolism   microbiology   MeSH
• Poultry Diseases metabolism   pathology   MeSH
• Salmonella enteritidis physiology   MeSH
• Salmonella Infections, Animal metabolism   physiopathology   MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Transcriptome * MeSH
• Up-Regulation MeSH
• Animals MeSH
• Check Tag
• Chick Embryo MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Burkholderia cenocepacia causes severe pulmonary infections in cystic fibrosis (CF) patients. Since the bacterium is virtually untreatable by antibiotics, chronic infections persist for years and might develop into fatal septic pneumonia (cepacia syndrome, CS). To devise new strategies to combat chronic B. cenocepacia infections, it is essential to obtain comprehensive knowledge about their pathogenesis. We conducted a comparative genomic analysis of 32 Czech isolates of epidemic clone B. cenocepacia ST32 isolated from various stages of chronic infection in 8 CF patients. High numbers of large-scale deletions were found to occur during chronic infection, affecting preferentially genomic islands and nonessential replicons. Recombination between insertion sequences (IS) was inferred as the mechanism behind deletion formation; the most numerous IS group was specific for the ST32 clone and has undergone transposition burst since its divergence. Genes functionally related to transition metal metabolism were identified as hotspots for deletions and IS insertions. This functional category was also represented among genes where nonsynonymous point mutations and indels occurred parallelly among patients. Another category exhibiting parallel mutations was oxidative stress protection; mutations in catalase KatG resulted in impaired detoxification of hydrogen peroxide. Deep sequencing revealed substantial polymorphism in genes of both categories within the sputum B. cenocepacia ST32 populations, indicating extensive adaptive evolution. Neither oxidative stress response nor transition metal metabolism genes were previously reported to undergo parallel evolution during chronic CF infection. Mutations in katG and copper metabolism genes were overrepresented in patients where chronic infection developed into CS. Among professional phagocytes, macrophages use both hydrogen peroxide and copper for their bactericidal activity; our results thus tentatively point to macrophages as suspects in pathogenesis towards the fatal CS.

• MeSH
• Burkholderia cenocepacia genetics   MeSH
• Chronic Disease MeSH
• Cystic Fibrosis complications   microbiology   MeSH
• Burkholderia Infections genetics   MeSH
• Respiratory Tract Infections microbiology   MeSH
• Humans MeSH
• Comparative Genomic Hybridization MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: The aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections. METHODS: For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 10⁹ CFU pro toto). RESULTS: Subcutaneous inoculation of a single dose with 2.6 × 10⁹ colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation. CONCLUSIONS: Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.

• MeSH
• Time Factors MeSH
• Francisella tularensis MeSH
• Thiobarbituric Acid Reactive Substances MeSH
• Oxidative Stress MeSH
• Serum Albumin metabolism   MeSH
• Tularemia metabolism   pathology   veterinary   MeSH
• Hares MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: More than 90% of cases of anal cancers are caused by high-risk human papillomavirus (HR HPV) infection and a history of cervical intraepithelial neoplasia (CIN) is established as possible risk factor. OBJECTIVES: To demonstrate relationship between anal and cervical HPV infection in women with different grades of CIN and microinvasive cervical cancer. STUDY DESIGN: A total of 272 women were enrolled in the study. The study group included 172 women who underwent conization for high-grade CIN or microinvasive cervical cancer. The control group consisted of 100 women with non-neoplastic gynecologic diseases or biopsy-confirmed CIN 1. All participants completed a questionnaire detailing their medical history and sexual risk factors and were subjected to anal and cervical HPV genotyping using Cobas and Lynear array HPV test. RESULTS: Cervical, anal, and concurrent cervical and anal HPV infections were detected in 82.6%, 48.3% and 42.4% of women in the study group, and in 28.0%, 26.0% and 8.0% of women in the control group, respectively. The prevalence of the HR HPV genotypes was higher in the study group and significantly increased with the severity of cervical lesion. Concurrent infections of the cervix and anus occurred 5.3-fold more often in the study group than in the control group. Any contact with the anus was the only significant risk factor for development of concurrent HPV infection. CONCLUSIONS: Concurrent anal and cervical HR HPV infection was found in nearly half of women with CIN 2+. The dominant genotype found in both anatomical locations was HPV 16. Any frequency and any type of contact with the anus were shown as the most important risk factor for concurrent HPV infection.

• MeSH
• Anal Canal virology   MeSH
• Cervix Uteri virology   MeSH
• Adult MeSH
• Uterine Cervical Dysplasia virology   MeSH
• Genotype MeSH
• Papillomavirus Infections complications   epidemiology   transmission   virology   MeSH
• Cohort Studies MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Molecular Epidemiology MeSH
• Papillomaviridae classification   genetics   isolation & purification   MeSH
• Surveys and Questionnaires MeSH
• Sexual Behavior MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Using high-throughput analyses and the TRANSFAC database, we characterized TF signatures of head and neck squamous cell carcinoma (HNSCC) subgroups by inferential analysis of target gene expression, correcting for the effects of DNA methylation and copy number. Using this discovery pipeline, we determined that human papillomavirus-related (HPV+) and HPV- HNSCC differed significantly based on the activity levels of key TFs including AP1, STATs, NF-κB and p53. Immunohistochemical analysis confirmed that HPV- HNSCC is characterized by co-activated STAT3 and NF-κB pathways and functional studies demonstrate that this phenotype can be effectively targeted with combined anti-NF-κB and anti-STAT therapies. These discoveries correlate strongly with previous findings connecting STATs, NF-κB and AP1 in HNSCC. We identified five top-scoring pair biomarkers from STATs, NF-κB and AP1 pathways that distinguish HPV+ from HPV- HNSCC based on TF activity and validated these biomarkers on TCGA and on independent validation cohorts. We conclude that a novel approach to TF pathway analysis can provide insight into therapeutic targeting of patient subgroup for heterogeneous disease such as HNSCC.

• MeSH
• Papillomavirus Infections genetics   metabolism   MeSH
• Humans MeSH
• DNA Methylation MeSH
• Cell Line, Tumor MeSH
• Head and Neck Neoplasms genetics   metabolism   virology   MeSH
• NF-kappa B genetics   metabolism   MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Signal Transduction MeSH
• Carcinoma, Squamous Cell genetics   metabolism   virology   MeSH
• STAT3 Transcription Factor genetics   metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The role of infection in autoimmunity is widely discussed. In this study we concentrated on relationship between HELICOBACTER PYLORI as a very important gastroduodenal pathogen and autoimmune thyroiditis (AT). Forty seven AT patients and 34 healthy controls were enrolled. They were split into: THP ( H.PYLORI positive patients, n=17), THN ( H.PYLORI negative patients, n=30), CP ( H.PYLORI positive controls, n=17) and CN groups ( H.PYLORI negative controls, n=17). By protein microarray we analysed production of 23 cytokines and chemokines prior and post stimulation with H.PYLORI lysate and its lipopolysaccharide (LPS). Reactivity to lysate as well as to bacterial LPS differed within groups. The lowest basal cytokine and chemokine production was observed in CN group but these subjects reacted significantly to specific stimulation by increasing IFN-gamma (in comparison with THP p=0.01 for LPS and p=0.004 for H.PYLORI lysate) and TGF-beta production (p=0.015 for LPS). In contrast, IL-10 and IL-5 were decreased in this group. In CP, THN and THP groups, we observed in general higher chemokine response. THP group increased proinflammatory IL-6 after specific stimulation as well (in comparison with CP p<0.0001 for LPS stimulation). We observed different "reactivity pattern" to H.PYLORI within groups with low basal cytokine and chemokine production in healthy H.PYLORI negative controls but with clear specific response in IFN-gamma and TGF-beta production in this group. Adequate immune reaction which is joined to appropriate immunoregulation leads to prevention of the chronic infection and on the other hand may prevent the development of "connected" diseases such as autoimmune. J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart.New York.

• MeSH
• Thyroiditis, Autoimmune immunology   microbiology   MeSH
• Chemokines biosynthesis   MeSH
• Protein Array Analysis MeSH
• Cytokines biosynthesis   MeSH
• Child MeSH
• Adult MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Helicobacter pylori immunology   MeSH
• Helicobacter Infections immunology   microbiology   MeSH
• Cells, Cultured MeSH
• Leukocytes, Mononuclear immunology   MeSH
• Humans MeSH
• Adolescent MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Humans MeSH
• Adolescent MeSH
• Female MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

OBJECTIVES: Umbilical cord blood (UCB) has become a useful alternative source of hematopoietic stem cells for clinical and research applications. UCB represents neonatal blood and differs from adult blood in many aspects, displaying different cell composition and various features of cellular immaturity. To understand molecular basis of phenotypic differences between neonatal and adult blood, we studied variations in transcriptome of UCB and maternal peripheral blood (PB). METHODS: Using Illumina microarrays, we determined gene expression profiles of UCB and PB samples obtained from 30 mothers giving birth to living baby. RESULTS: Out of 20,589 tested genes, 424 genes were down-regulated and 417 genes were up-regulated in UCB compared with PB. Reduced expression of many immunity-related pathways (e.g. TLR pathway, Jak-STAT pathway, cytokine-cytokine receptor interaction) in neonatal blood cells may contribute to the poor response to antigens, increasing susceptibility to infections at the time of disappearance of protective maternal antibodies. On the other hand, overexpression of erythropoiesis-related genes (glycophorins, fetal hemoglobins, enzymes catalysing heme synthesis and erythrocyte differentiation) in UCB probably enforces red cell production in newborns. CONCLUSIONS: Our study demonstrates that neonatal and maternal bloods show specific gene expression profiles, likely reflecting differences in phenotypes of immunologically immature and fully evolved hematopoietic cells.

• MeSH
• Phenotype MeSH
• Fetal Blood cytology   MeSH
• Immune System MeSH
• Humans MeSH
• Mothers MeSH
• Infant, Newborn MeSH
• Umbilical Cord pathology   MeSH
• Gene Expression Regulation MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Cluster Analysis MeSH
• Gene Expression Profiling methods   MeSH
• Pregnancy MeSH
• Pregnancy Outcome MeSH
• Check Tag
• Humans MeSH
• Infant, Newborn MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

Gene duplication is an important evolutionary mechanism and no eukaryote has more duplicated gene families than the parasitic protist Trichomonas vaginalis. Iron is an essential nutrient for Trichomonas and plays a pivotal role in the establishment of infection, proliferation, and virulence. To gain insight into the role of iron in T. vaginalis gene expression and genome evolution, we screened iron-regulated genes using an oligonucleotide microarray for T. vaginalis and by comparative EST (expressed sequence tag) sequencing of cDNA libraries derived from trichomonads cultivated under iron-rich (+Fe) and iron-restricted (-Fe) conditions. Among 19,000 ESTs from both libraries, we identified 336 iron-regulated genes, of which 165 were upregulated under +Fe conditions and 171 under -Fe conditions. The microarray analysis revealed that 195 of 4,950 unique genes were differentially expressed. Of these, 117 genes were upregulated under +Fe conditions and 78 were upregulated under -Fe conditions. The results of both methods were congruent concerning the regulatory trends and the representation of gene categories. Under +Fe conditions, the expression of proteins involved in carbohydrate metabolism, particularly in the energy metabolism of hydrogenosomes, and in methionine catabolism was increased. The iron-sulfur cluster assembly machinery and certain cysteine proteases are of particular importance among the proteins upregulated under -Fe conditions. A unique feature of the T. vaginalis genome is the retention during evolution of multiple paralogous copies for a majority of all genes. Although the origins and reasons for this gene expansion remain unclear, the retention of multiple gene copies could provide an opportunity to evolve differential expression during growth in variable environmental conditions. For genes whose expression was affected by iron, we found that iron influenced the expression of only some of the paralogous copies, whereas the expression of the other paralogs was iron independent. This finding indicates a very stringent regulation of the differentially expressed paralogous genes in response to changes in the availability of exogenous nutrients and provides insight into the evolutionary rationale underlying massive paralog retention in the Trichomonas genome.

• MeSH
• Cysteine Proteases genetics   metabolism   MeSH
• Gene Duplication MeSH
• Expressed Sequence Tags MeSH
• Genome, Protozoan MeSH
• Gene Dosage MeSH
• Gene Library MeSH
• Glycolysis genetics   MeSH
• Evolution, Molecular MeSH
• Iron-Sulfur Proteins genetics   metabolism   MeSH
• Genes, Protozoan * MeSH
• Gene Expression Regulation * MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Transcriptome * MeSH
• Trichomonas vaginalis genetics   metabolism   MeSH
• Iron metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH