23 research outputs found

    The Role of the County Professional Council in Advanced Training and Professional Development of Class Teachers

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    Stručno usvršavanje je obvezan dio učiteljskog posla koji se provodi na četiri osnovne razine: individualnoj, školskoj, županijskoj i državnoj. Brojne su prednosti i nedostaci svakog oblika usavršavanja, a svi su oni na putu profesionalnog razvoja učitelja jednako važni i korisni. U radu je prikazan model stručnog usvršavanja učitelja na županijskoj razini (Županijsko stručno vijeće učitelja razredne nastave – Grad Sisak) pri čemu veliku ulogu imaju upravo županijski voditelji koji su poveznica između školske i državne razine stručnog usavršavanja učitelja.Advanced training is a mandatory part of teacher’s job and it is being carried out at four basic levels: individual, school, county and state level. There are numerous advantages and disadvantages of any form of training, and they are all equally important and useful in the course of professional development of teachers. This study presents the model of advanced training of class teachers at county level (County professional council of class teachers – the Town of Sisak), where the very county leaders, who are the link between the school and state level of advanced training of teachers, have a great role

    Restriction enzyme analyses of <i>B</i>. <i>pseudomallei</i> phages.

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    <p>Genomic DNA extracted from soil isolated (ΦBp-RE4-5) and MMC-induced (ΦBp-RE1-3) <i>B</i>. <i>pseudomallei</i> phages were digested with restriction enzyme <i>Bst</i>BI (A) or <i>Mlu</i>I (B) and analyzed using agarose gel electrophoresis. Different DNA patterns were observed when digested with the <i>Mlu</i>I restriction enzyme. A 1-kb DNA ladder was included as a DNA marker.</p

    PCR amplification and DNA sequence analysis of the DNA fragment encoding the <i>B</i>. <i>pseudomallei</i> phage tail tubular protein B.

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    <p>DNA from soil-isolated and MMC-induced temperate phages were used as a template to PCR amplify the fragment of a gene encoding the phage tail tubular protein B. The 325-bp DNA fragment was detected in each case. A negative PCR control (N) and 100-bp DNA marker were included (A). Nucleotide sequences comparison of the phage tail tubular protein B DNA amplified from ΦBp-RE1 and ΦBp-AMP1 is shown (B).</p

    Confocal micrographs indicating <i>bsaQ</i>-dependent secretion of CHBP in U937 cells infected with <i>B. pseudomallei</i>.

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    <p>PMA-activated U937 cells were separately infected with <i>B. pseudomallei</i> (K96243, <i>bsaQ</i> or <i>chbP</i> mutant strain). At different time points of infection (3, 6, 9 and 12 h), infected cells were stained using purified rabbit anti-CHBP antibody detected with anti-rabbit Ig-Alexa Fluor<sup>488</sup> (Molecular Probes). The bottom panel shows the localization of CHBP and the top panel merges this signal with DIC images showing the position of infected cells. Scale bars, 10 µm.</p

    SDS-PAGE and Western blot analysis of CHBP in <i>B. pseudomallei</i> K96243 wild-type, <i>chbP</i> mutant and <i>trans</i>-complemented strains.

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    <p>A) SDS-PAGE. Bacterial lysates and secreted proteins of <i>B. pseudomallei</i> K96243, <i>chbP</i> mutant or <i>chbP</i>/pCHBP strain cultured in LB broth for 6 h were separated by 12% polyacrylamide gel electrophoresis. B) Western blot analysis. The blotted proteins from A) were separately probed with anti-CHBP and anti-BopE antibodies. Molecular mass markers are shown on the left. Lanes 1–3 are bacterial cell lysates and lanes 4–6 are secreted proteins precipitated from culture supernatants.</p

    Effect of <i>chbP</i> mutation on <i>B. pseudomallei</i> plaque formation.

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    <p>A) Plaque-forming efficiency. HeLa cells were infected with <i>B. pseudomallei</i> (K96243, <i>chbP</i> mutant or <i>chbP</i>/pCHBP strain) at an MOI of 20. Plaque-forming efficiency was established following staining of the infected cells with crystal violet. Plaque-forming efficiency at 21 h was calculated by the following equation: number of plaques/CFU of bacteria added per well. Asterisks indicate significant differences (P value <0.05, <i>t</i>-test) between groups. Error bars represent standard errors of the means for experiments performed in triplicate. B) Photographs of plaques. Representative images of the infected cell monolayers after infection with <i>B. pseudomallei</i> K96243, <i>chbP</i> mutant or <i>chbP</i>/pCHBP strains for 21 and 24 h. Note the reduced number of plaques and reduced plaque size of the <i>chbP</i> mutant.</p

    The growth of mitomycin C (MMC)-treated <i>B</i>. <i>pseudomallei</i> cultures and Transmission Electron Microscopy assessment of the induced phages.

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    <p>Growth of different strains following MMC induction. The MMC-treated cultures show a decline in optical density (OD<sub>600nm</sub>) after addition of MMC that is not observed in the untreated culture (A). Soil-isolated (B) and MMC-induced temperate (C) phages have similar icosahedral heads with short, non-contractile tails, which are characteristic of phages in the <i>Podoviridae</i> family.</p

    Effect of <i>chbP</i> mutation on <i>B. pseudomallei-</i>induced cytotoxicity.

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    <p>A) HeLa cells and B) U937 cells were infected with <i>B. pseudomallei</i> (K96243, <i>chbP</i> mutant or <i>chbP</i>/pCHBP strain) with a range of MOIs. After 6 h, cytotoxicity was assessed using the CytoTox96 lactate dehydrogenase (LDH)-release kit (Promega). Asterisks indicate significant differences (P value <0.05, <i>t</i>-test) between groups. Error bars represent standard errors of the means for experiments performed in triplicate.</p

    Confocal micrographs of CHBP expression and localization in U937 cells infected with <i>B. pseudomallei</i>.

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    <p>PMA-activated U937 cells were separately infected with three strains of <i>B. pseudomallei</i> (K96243, <i>chbP</i> mutant or <i>chbP</i>/pCHBP strain). After 6 h, infected cells were fixed, permeabilized andstained using purified rabbit anti-CHBP antibody detected with anti-rabbit Ig-Alexa Fluor<sup>488</sup> (Molecular Probes). The bottom panel shows the localization of CHBP and the top panel merges this signal with differential interference contrast (DIC) images showing the position of infected cells. Scale bars, 20 µm.</p

    SDS-PAGE and Western blot analysis of CHBP in <i>B. pseudomallei-</i>infected U937 cells.

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    <p>A) Protein from lysates of U937 cells infected at an MOI of 2 with <i>B. pseudomallei</i> K96243, <i>chbP</i> mutant, <i>chbP</i>/pCHBP strain or <i>bsaQ</i> mutant for 3 or 6 h were separated by 12% polyacrylamide gel electrophoresis and blotted with anti-CHBP and anti-BopE antibodies. Molecular mass markers are shown on the left. Panel B shows data from an identical experiment, except using an MOI of 100.</p
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