«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

Validation of microarray data using multiple <i>B. pertussis ptxP1</i> and <i>ptxP3</i> strains.

<p>Sulfate was added to the culture medium to induce high (50 mM), medium (5 mM), and low (<0.02 mM, represented as 0 mM) sulfate conditions. (A) qRT-PCR data showing the relative expression level of <i>vag8</i>, <i>lpxE, sbp, cysB, ptxA, prn,</i> and <i>fhaB</i> between <i>ptxP1</i> strains (represented by white symbols) and <i>ptxP3</i> strains (represented by black symbols) grown under low (0 mM), medium (5 mM) and high (50 mM) sulfate conditions. The data are expressed as fold changes relative to the <i>ptxP1</i> strains grown under low sulfate conditions, with horizontal bars representing the geometrical mean, so that any sulfate-dependent effects on gene expression become apparent. B) Luminex data showing Ptx, Prn, and FHA protein expression in <i>ptxP3</i> and <i>ptxP1</i> strains grown under low (0 mM), medium (5 mM) and high (50 mM) sulfate conditions. Protein expression is expressed in absolute amounts (nanograms) with horizontal bars representing the geometrical mean. The expression values of the two strains that were used for microarray analysis (B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>)) are depicted in grey. Asterisks indicate a statistically significant difference between the groups as determined by an unpaired Student's t-test: * <i>P</i> value <0.05, ** <i>P</i> value <0.005, *** <i>P</i> value <0.0005.</p

Sulfate-mediated modulation in <i>B. pertussis</i> strain B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>).

<p>Sulfate was added to the culture medium to induce a high (50 mM), medium (5 mM), and low (<0.02 mM, represented as 0 mM on the x-axis) sulfate conditions. qRT-PCR data shows the relative expression level of <i>kpsT</i>, <i>bipA</i>, and <i>ptxA</i> expressed as fold changes relative to the high sulfate condition, with the values being the mean of four biological replicate cultures. Asterisks indicate a statistically significant difference between the groups as determined by Student's t-test with Welch's correction: * <i>P</i> value <0.05, ** <i>P</i> value <0.005, *** <i>P</i> value <0.0005.</p

Genes differentially regulated between <i>B. pertussis</i> strain B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>) upon sulfate-modulation.

<p>Sulfate was used to induce high (50 mM), medium (5 mM), and low (<0.02 mM, represented as 0 mM) sulfate conditions in a <i>ptxP3</i> and <i>ptxP1</i> strain. A) The number of genes expressed at least 3-fold higher between strains are indicated for each sulfate concentration. B&C) The pie charts subdivide the 44 genes expressed higher in the <i>ptxP1</i> strain as compared to the <i>ptxP3</i> strain under high sulfate conditions (B) and the 61 genes expressed higher in the <i>ptxP3</i> strain as compared to the <i>ptxP1</i> strain under medium sulfate conditions (C) into functional gene categories. The number of genes belonging to each category is numbered between brackets.</p

Functional categorization of sulfate-regulated genes in <i>B. pertussis</i> strain B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>).

<p>Venn diagrams show overlapping and unique sulfate-regulated genes that are at least 3-fold downregulated under high sulfate conditions relative to the low sulfate condition (A; high sulfate repressed (HSR)), 3-fold upregulated under high sulfate conditions relative to the low sulfate condition (D; high sulfate induced (HSI)), 3-fold up- or down-regulated under medium sulfate conditions relative to the low sulfate condition (medium sulfate induced (MSI) and repressed (MSR)). HSR, HSI, and MSR genes were grouped by functional categories (B, E, and H) and PSORTb-predicated subcellular localization (C, F, and I). Data are expressed as the percentage that is sulfate-regulated among all annotated genes in each class. Asterisks indicate statistically significant enrichment of sulfate-regulated genes in a certain class as determined by Fisher's exact test. * <i>P</i> value <0.05, ** <i>P</i> value <0.005, *** <i>P</i> value <0.0005.</p

Medium-sulfate regulated genes in <i>B. pertussis</i> strain B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>).

<p>Abbreviations; C, cytoplasmic; Cm, cytoplasmic membrane; E, extracellular; FC, fold change; MSR, medium sulfate repressed; Om, outer membrane; ORF, open reading frame; P, periplasmic; Un, unknown. The columns with fold change difference show the ratio of absolute expression (<i>ptxP3</i>/<i>ptxP1</i>) under low (0 mM), intermediate (5 mM), and high (50 mM) sulfate conditions. * Asterisks indicate a statistically significant difference. ** indicate a fold-change and statistically significant difference as determined by qPCR (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084523#pone-0084523-g001" target="_blank">Figure 1</a>). † indicates that the gene was excluded because the microarray signal intensity was ≤500 in both the conditions compared.</p

Transcriptional regulation of sulfate genes in <i>B. pertussis</i> strain B1920 (<i>ptxP1</i>) and B1917 (<i>ptxP3</i>).

<p>Abbreviations; C, cytoplasmic; Cm, cytoplasmic membrane; FC, fold change; HSR, high sulfate repressed; ORF, open reading frame; P, periplasmic; Un, unknown. The columns with fold change difference show the ratio of absolute expression (<i>ptxP3</i>/<i>ptxP1</i>) under low (0 mM), intermediate (5 mM), and high (50 mM) sulfate conditions. Asterisks indicate a statistically significant difference. † indicates that the gene was excluded because signal intensity was ≤500 in both the conditions compared. Genes above the dotted line are genes that annotated as genes involved in sulfate metabolism.</p

Characteristics of the Dutch <i>Bordetella pertussis</i> strains used in this study.

<p>Characteristics of the Dutch <i>Bordetella pertussis</i> strains used in this study.</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