28 research outputs found
Neonatal anthropometry: thin-fat phenotype in fourth to fifth generation south Asian neonates in Surinam
No full textWe assessed whether the earlier described 'thin-fat phenotype' is present in Surinam South Asian babies of the fourth to fifth generation after migration from India. In this observational study we collected data from 39 South Asian term neonates and their mothers in Paramaribo, Surinam. We compared the following data with data from an earlier study in Southampton, UK (338 neonates) and in Pune, India (631 neonates): maternal body mass index, neonatal weight, length, head, mid-upper arm and abdominal circumferences and subscapular skinfold thickness. The mothers in Paramaribo were older than the Southampton mothers; their body mass index was comparable. Mean birth weight was 3159 g (Southampton: 3494 g; Pune: 2666 g). Compared with Southampton babies, the Paramaribo babies were smaller in nearly all body measurements, the smallest being abdominal circumference at the umbilicus level (s.d. score: -1.62; 95% confidence interval (CI): -2.07 to -1.16) and mid-upper arm circumference (s.d. score: -1.08; 95% CI: -1.46 to -0.69). In contrast, subscapular skinfold thickness was similar (s.d. score: +0.08; 95% CI: -0.24 to +0.55). Except for subscapular skinfold thickness and length, all neonatal measurements were intermediate between those from Southampton and Pune. The thin-fat phenotype is preserved in Surinam South Asian neonates of the fourth to fifth generation after migration from Indi
Mipu1 Inhibits Lipid Accumulation Through Down-Regulation of CD36 in RAW264.7 Cells
No full textBackground/Aims: Our recent data indicated that Mipu1 overexpression reduces lipid intake and CD36 expression of macrophages in the presence of oxLDL. However, the mechanism of Mipu1 inhibiting lipid accumulation in macrophages is not elucidated. Methods: Real-time quantitative polymerase chain reaction (PCR) and western blot analysis were used to detect expression of Mipu1 and CD36. The promoter activity of CD36 was studied using luciferase assays. Chromatin immunoprecipitation (ChIP) was used to show the recruitment of Mipu1 onto the CD36 promoter. High-performance liquid chromatography and Dil-labeled lipoprotein were used to detect cholesterol accumulation. Results: Here, we show that CD36 overexpression rescues oxLDL-induced cholesterol accumulation in RAW264.7-Mipu1 cells. Analysis of the mouse CD36 promoter revealed two potential Mipu1-response elements (MRE), one of which (from -237bp to -244bp, ACTTAC) was shown, using mutagenesis and deletion analysis, to be functional. Mipu1 was demonstrated to bind to CD36 promoter, and oxLDL treatment resulted in increases in their interaction as assessed by ChIP. Conclusions: It was demonstrated that Mipu1 inhibited the lipid accumulation of macrophages and it down-regulated CD36 expression in the presence of oxLDL
Contribuições de uma estratégia de avaliação articulando a análise escrita e o discurso oral de estudantes do ensino médio
No full textAs estratégias de avaliação da aprendizagem e as estratégias de ensino são múltiplas e permitem reconhecer de diferentes modos os conhecimentos construídos pelos estudantes. Neste trabalho buscou-se verificar as contribuições de uma estratégia de avaliação baseada no reconhecimento dos níveis cognitivos, combinando a análise da escrita, realizada através de questionários e do discurso oral, através de uma atividade denominada júri químico em uma sequência de ensino. Nossos dados apontam que a estratégia combinada permite reconhecer conhecimentos distintos construídos através de linguagens e formas de expressão diferentes utilizadas pelos alunos e possibilita aos alunos buscarem outros meios de acesso a informações e construção dos conhecimentos
Relative expression of <i>B. bigemina CCp1-3</i> by extracellular sexual stages incubated either at 37°C or 28°C for 24 h and 48 h is compared to gene expression by intracellular parasites (Intracell) incubated under identical conditions for each temperature and time point.
No full text<p>Gene expression was normalized to the total amount of RNA used to generate the cDNA. The transcription level was calculated as a relative expression using the formula: Relative expression <sub>(sample)</sub> = 2 <sup>[C<i>q</i> (control) – C<i>q</i> (sample)]</sup>, where the control is the highest C<i>q</i> value for a given gene of interest. Results represent the means of three experiments, each containing three technical replicates. Differences in gene expression were determined by the Student’s <i>t</i>-test. “a” indicates the expression of <i>CCp1</i>-<i>3</i> was significantly higher (P<0.001) by sexual stages incubated for 24 h at 37°C than by Intracell. “b” indicates the expression of <i>CCp1</i>and <i>CCp2</i> was significantly higher (P<0.001) by sexual stages incubated for 24 h at 28°C than by Intracell. “c” indicates the expression of <i>CCp1</i>and <i>CCp2</i> was significantly higher (P<0.001) by sexual stages incubated for 37 h at 48°C than by Intracell.</p
Immunoblot analysis demonstrating the expression of CCp1-3 by temperature-induced <i>B. bigemina</i> parasites.
No full text<p>Expression of CCp1-3 proteins was investigated in <i>B. bigemina</i> cultures incubated for 24 h at 28°C. Expression of CCp1 was detected only by extracellular sexual stages (Sex Stg) whereas CCp2 and CCp3 were detected in both Sex Stg and intracellular parasites (Intracell) populations. No bands with the expected molecular mass of the CCp proteins were detected in non-induced <i>B. bigemina</i> cultures (uCulture) or in uninfected red blood cells (uRBC). Polyclonal mouse anti-CCp1 peptide, rabbit anti-CCp2-peptides or CCp3-peptides, and bovine serum from an animal chronically infected with <i>B. bigemina</i> were used at a 1∶50 dilution. Anti-mouse, anti-rabbit, or anti-bovine HRP conjugates were used at a 1∶4,000 dilution. Black arrows indicate the expected size of CCp1-3.</p
The <i>gfp-bsd</i> and <i>MSA-1-Bm86ep</i> transgenes are co-expressed in transfected parasites.
No full text<p><b>A–B</b> Are the results from immunoblots using GFP (A) and Bm86 (B) specific antibodies. Lanes 1–5 represent recombinant MSA-1-BM86ep, recombinant GFP-BSD, whole cell lysate from the Tf-Bm86ep-gfp-bsd parasite line, whole cell lysate from Mo7 wild type, and non-infected RBC lysate control respectively. Molecular masses are indicated on the left of each panel. <b>C–J</b> Show results of fixed cell immunofluorescence using <i>B. bovis</i> Mo7 (C–F) and Tf-Bm86ep-gfp-bsd (G–J) infected erythrocytes from cultured cell lines. Parasites were stained with DAPI nucleic acid stain (C–G), GFP antibody labeled with Alexa Flour 488 (D–H), and Bm86ep antibody labeled with Alexa Flour 555 (E–I). Images were visualized using epifluorescence microscopy for each label, and a merged image was created (F–J). A 10 micron size bar is included in the bottom right panel.</p
Bm86 epitopes are expressed on the surface of Tf-Bm86ep-gfp-bsd extraerythrocytic merozoites.
No full text<p><b>A–D</b> represent permeabilized extraerythrocytic merozoites stained with DAPI (A), incubated with Bm86ep antibody labeled with Alexa Flour 647 (B), and GFP antibody labeled with Alexa Flour 488 (C). A merged image of panels B and C is shown in panel D. <b>E–H</b> represents identical staining procedures applied to non-permeabilized cells. <b>I–L</b> represent non-permeabilized extraerythrocytic merozoites stained with DAPI (I), and incubated with Bm86ep antibody labeled with Alexa Flour 647 (J), and the MSA-1 mAb Babb35 labeled with Alexa Flour 488 (K). A merged image of panels J and K is shown in panel L. A two micron size bar is included on the bottom right panel.</p
Expression of <i>CCp1</i>-<i>3</i> during acute <i>B. bovis</i> infection.
No full text<p>In panel A, relative fold expression of <i>CCp1-3</i> transcripts in bovine blood during acute <i>B. bovis</i> infection is shown in the right y-axis. Absolute quantification of <i>B. bovis msa-1</i> DNA in bovine blood is shown in the left y-axis. <i>CCp</i> gene expression was normalized to the total amount of RNA used to generate the cDNA and the transcription level was calculated as a relative expression using the formula: Relative expression <sub>(sample)</sub> = 2 <sup>[C<i>q</i> (control) – C<i>q</i> (sample)]</sup>, where the control is the highest C<i>q</i> value for a given gene of interest. To confirm the presence of amplifiable cDNA in all tested samples a standard PCR for bovine actin was performed. In panel B, absolute expression of <i>B. bovis msa-1</i> RNA in bovine blood is shown in the right y-axis and the percentage of engorged ticks infected with <i>B. bovis</i> is shown in the left y-axis.</p
Gene identification, cDNA length, protein length, expected protein size and schematic domain representation of CCp1-3 in <i>Babesia bovis</i> and <i>Theileria equi</i>.
No full text<p>Black lines represent the full-length of the CCp proteins in <i>B. bovis</i> and <i>T. equi</i>. Grey lines represent the regions of the CCp proteins available in <i>B. bigemina</i>. Red lines above CCp1-3 indicate the regions used to design synthetic peptides. R: Ricin B lectin domain. F: F5/F8 type C domain. L: LCCL domain. PL: PLAT domain. SR: Scavenger receptor domain. * <i>In silico</i> data of <i>B. bovis</i> CCp1-3 has been previously shown by Becker <i>et a</i>l <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067765#pone.0067765-Becker1" target="_blank">[11]</a>.</p
Full size <i>B. bovis ef-1α</i> IG region acts as bidirectional promoter.
No full text<p><b>A</b>. Plasmids <i>pLuc</i> (no promoter control), <i>pEfIgAluc</i> (one functional luciferase gene under transcriptional control of the Ig-A promoter), <i>pEfIgBluc</i> (one functional luciferase gene under transcriptional control of the Ig-B promoter), and <i>pEfIgA&Bluc</i> (two functional luciferase genes under transcriptional control of the Ig-A and Ig-B promoters) are schematically represented together with a chart showing luciferase activity generated by transiently transfected Mo7 <i>B. bovis</i>. The 1.4 kb <i>ef-1α</i> IG is shown in yellow with “A” and “B” orientation of the promoter as indicated with an arrow. The luciferase genes are represented in red and the <i>3′ rap</i> region used to stop transcription is indicated in grey. Data on the right represent luciferase expression in <i>B. bovis</i> lysates at 24 hours after transfection from each respective transfection plasmid. Luciferase activity is expressed as relative luciferase units (RLU). Error bars represent standard deviation of the mean for three independent experiments. Asterisk represents significant differences among luciferase activities on lysates from parasites transfected with plasmids containing a promoter. (<i>p</i><0.05). <b>B</b>. Percentage of parasitized erythrocytes (PPE) of transfected parasite lines transfected with <i>pEfIgAluc, pEfIgBluc</i>, and <i>pEfIgA&Bluc</i>, obtained at 4 and 24 hours post-electroporation. Error bars represent standard deviation of the mean for three independent experiments. <b>C</b>. Quantification of the number of plasmid copies per MSA-1 gene copy (luc/genome copy), considering that the <i>B. bovis</i> genome contains a single MSA-1 gene, calculated 24 hours post-electroporation using quantitative PCR. Each cell line transfected with <i>pEfIgAluc, pEfIgBluc</i>, and <i>pEfIgA&Bluc</i> are shown. Error bars represent standard deviation of the mean for three independent experiments.</p
