«Високе» і «низьке» у творчості народній і писемній

«The high» and «the low», – these categories can be examined and as mythical, as poetic image of space, as metaphorical recreation of human fate, as a public vertical line of human possibilities, as the valued step at the analysis of artistic styles and at the same time as violation of such evaluation. All these aspects adds to the theme as as an object of attention in the article; the relations of «high» and «low» culture, culture folk comes forward and «lordly» in the past centuries. Categories highly/low, higher/below are examined in various scientific studios – mythological, culturological, sociological, from the point of view of theory of literature, in historical and literary measuring.Visoko i nisko, gore i dolje može se promatrati kao mitološko značenje, kao poetska vizija prostora, kao metafora ljudske sudbine, kao društvena vertikala moći, kao vrijednosna ljestvica književnih stilova, kao narušavanje takvoga vrednovanja. Sve su to tek dopune temi kojom se želimo baviti. Zanimljivi su za nas odnosi visoke i niske kulture (prvenstveno verbalne, književne), kulture pučke i gospodske u proteklim stoljećima. Pojmovi se visoko/nisko, gore/dol

Additional file 1: Table S1. of A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination

Silent mutations introduced in rIBV. Nucleotide positions and sequences refer to the IBV H52 BI genome (see Additional file 2: Figure S1). Modified nucleotides in recombinant IBV wt are depicted in lower case; n.a. = not applicable. Purpose of introduction of restriction enzyme site are indicated for each site; in case of enzyme site removal the purpose was to create unique restriction enzyme sites for cloning; n.a. = not applicable in this study. (DOCX 19 kb

Additional file 2: Figure S1. of A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination

Alignment of 3′ 9 kb of mIBV and rIBV-wt with IBV H52 BI. Alignment of the 3′ 9 kb of mIBV 1B3IIA P6 (excluding the MHV derived spike ectodomain sequence) and recombinant (r)IBV wild-type (wt) P4 with IBV H52 BI. Numbers refer to nucleotide positions in the IBV H52 BI genome. Restriction enzyme sites are highlighted in yellow, with the corresponding enzyme indicated above the sequences. An additional thymidine residue to keep the MHV spike gene ectodomain sequence in frame with the IBV spike gene signal sequence at position 20,385 is highlighted in green and marked with a # above the sequence. A spontaneous T to C silent substitution in the spike of rIBV-wt at position 22,644 is highlighted in red. (DOCX 41 kb

Effect of immunization with novel vaccine candidates on infection with <i>B. pertussis</i> in the lower and upper respiratory tract.

<p>Naïve adult female BALB/c mice were subcutaneously immunized as described in the text and infected intranasally with 2×10⁷ CFU of <i>B. pertussis</i> strain B1917. The bacterial load in the lungs and nose was quantified three (<b>A&C</b>) and seven (<b>B&D</b>) days after challenge. Each symbol represents one mouse. Horizontal lines represents the mean. Dashed lines indicate the lower limit of detection. *<i>p</i><0.05, **<i>p</i><0.005 relative to PBS mice; 2-tailed Mann-Whitney <i>U</i> test.</p

Vaccine antigen selection and functional clustering of Bvg-activated proteins.

<p><b>A</b>) Putative protein antigens were selected based on ≥2.5-fold Bvg-activation at both protein (this work) and mRNA level <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105011#pone.0105011-deGouw1" target="_blank">[10]</a>, presence in the core genome of <i>B. pertussis </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105011#pone.0105011-King1" target="_blank">[25]</a>, and PSORTb v3.0 predicted surface accessibility (outer membrane or extracellular) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105011#pone.0105011-Yu1" target="_blank">[26]</a>. This resulted in the identification of 15 putative vaccine candidates. † known <i>B. pertussis</i> protective antigens. Proteins in bold were selected for further examination. The proteins in bold The 855 and 51 proteins that were respectively identified and Bvg-activated at the protein level in both strains, were grouped by functional categories (<b>B</b>) and PSORTb-predicated subcellular localization (<b>C</b>). The relative frequency of Bvg-activated proteins compared to the total number of annotated proteins identified in both strains for each functional class are listed on the right-hand side. Asterisks indicate statistically significant enrichment of Bvg-activated proteins in a certain class as determined by Fisher's exact test. *<i>p</i><0.05, ***<i>p</i><0.0005.</p

<i>In vivo</i> expression of vaccine-candidate genes.

<p>Naïve adult female BALB/c mice (n = 4) were infected intranasally with <i>B. pertussis</i> strain B1917 or B1920. After 7 days, bacteria were collected through broncho-alveolar lavage and used for <i>in vivo</i> transcriptional analysis using antigen-specific primers as described in the text. The transcription data is expressed as 40-ΔCt value, which is a measure of expression relative to the <i>recA</i> household gene (ΔCt  =  Ct <i>target</i> – Ct <i>recA</i>). The number 40 represents the number of PCR cycles. A 40-ΔCt value of 40 indicates that the gene is expressed at equal levels as <i>recA</i>, while higher values correspond to higher expression.</p

Antibody-mediated opsonization of <i>B. pertussis</i>.

<p>The binding of serum (<b>A</b>) and nasal lavage (<b>B</b>) IgG antibodies to Bvg⁺ or Bvg⁻ grown <i>B. pertussis</i> B1917 was determined using flow cytometry. Bars represent the geometric mean ±95% confidence interval (CI) of 6 individual mice. *<i>p</i><0.05, ***<i>p</i><0.0005 relative to PBS group; Kruskal-Wallis test followed by a Dunns <i>post-hoc</i> test (α = 5%). To determine the distribution of IgG subtypes, whole-cell ELISA was performed with the post-immunization serum samples to detect pertussis-specific IgG1 (<b>C</b>), IgG2a (<b>D</b>), and IgG2b (<b>E</b>). Each symbol represents one mouse and the geometric mean is represented by a line. Dashed lines indicate the lower limit of detection (23, 4, and 27 ng/ml for IgG1, IgG2a, and IgG2b, respectively). For IgG1, a 1-tailed Wilcoxon Signed Rank Test was performed because IgG1 levels of the PBS mice were below the detection limit. IgG2a and IgG2b levels were statistically compared to the PBS mice using a 2-tailed Mann-Whitney <i>U</i>-test. *<i>p</i><0.05, **<i>p</i><0.005.</p

Detection of antigen-specific IgG in the serum of immunized mice.

<p>Post-immune serum (day 28) was used to determine the amount of total IgG specific for each recombinant antigens using ELISA as described in the text. Each symbol represents one mouse and the geometric mean is represented by a line. Dashed lines indicate the lower limit of detection (93 ng/ml). *<i>p</i><0.05; 1-tailed Wilcoxon Signed Rank Test.</p