Body mass index and blood pressure in a semi-urban community in Ota, Nigeria

This study was designed to establish the relationship between body mass index (BMI) and blood pressure (BP) in an increasingly industrialised town in Nigeria due to the rising prevalence of hypertension in non-industrialised countries. Factors associated with BMI and BP levels were determined in three hundred adult male and female subjects in Ota community of Ogun State, Nigeria. The levels of the overweight among the male and female subjects were 53.03 % and 47.37 % respectively. The levels of hypertensive male and female subjects were 40.91 % and 35.34 % respectively. The overweight and underweight among the hypertensive male were 54.29 % and 0 % respectively; while the overweight and underweight among the hypertensive female were 42.86 % and 28.57 % respectively. Hypertension among the overweight, and hypotension among the underweight, are major health concern in Ota that requires intensive medical care

Wild type <i>Shigella</i> increase Caco2 monolayer paracellular permeability to macromolecules whilst vaccine candidates have no or markedly attenuated effect.

<p><b>A</b>. FITC-BSA (66 kDa) net transport after infection with wild-type <i>S. flexneri</i> 2a or CVD 1208S applied at three different inocula. <b>B</b>. FITC-BSA (66 kDa) net transport after infection with wild-type <i>S. dysenteriae</i>-1 or CVD 1256 applied at three different inocula. C. FITC-Dextran (4 kDa) net transport after infection with wild-type filtered (Cond. Media: CM) or live <i>Shigella</i> applied at 10⁶ CFU. Calcium-free medium supplemented with EGTA to disrupt TJs served as positive control. Results are expressed as mean ± SEM of triplicate samples for each condition and are representative of 3 experiments with similar results. ***<i>p</i><0.001, *<i>p</i><0.05 compared to uninfected control (ANOVA).</p

Wild type <i>Shigella</i> induce TEER decrease in Caco2 cell monolayers independent of MOIs.

<p><b>A</b>. TEER responses to wild-type <i>S. flexneri</i> 2a applied at three different inocula. <b>B</b>. TEER responses to <i>S. flexneri</i> 2a heat-killed bacteria and culture supernatants (Cond. Media). <b>C</b>. TEER responses to wild-type <i>S. dysenteriae</i> 1 applied at three different inocula. <b>D</b>. TEER responses to <i>S. dysenteriae</i> 1 heat-killed bacteria and culture supernatants. Data are expressed as means ± SEM for triplicate samples for all conditions tested. * and # indicate statistically significant differences vs. t = 0 and vs. the uninfected control at the same time point, respectively; <i>p</i><0.05 (ANOVA). These results are representative of 3 experiments with similar results.</p

Occludin is hyperphosphorylated after infection with <i>Shigella</i> strains and disassembles from tight-junctions.

<p><b>A</b>, <b>B</b>. Western blot of protein samples in the form of X-100 Triton soluble (Sol) and insoluble (Ins) fractions at 6 hours post-infection, blotted with anti-occludin (upper rows), anti-phosphothreonine (middle rows) and anti-actin (lower rows). <b>C</b>, <b>D</b>. Western blot of protein samples at 24 hours post-infection, blotted with anti-occludin (upper rows), anti-phosphothreonine (middle rows) and anti-actin (lower rows).</p

Wild type <i>Shigella</i>, but not vaccine strains, moderately affect Caco2 monolayer viability.

<p><b>A</b>. LDH release after infection with wild-type <i>S. flexneri</i> 2a or its vaccine strain CVD 1208S at different inocula. <b>B</b>. LDH release after infection with wild-type <i>S. dysenteriae</i> 1 or its vaccine strain CVD 1256 applied at different inocula. Caco2 cells were infected with bacteria for 6 hours, washed and incubated overnight at 37°C. Apical supernatants were collected at 24 hours post-infection for LDH release measurement. Values represent LDH release from cells into the medium as a percentage of total LDH (Control). Data are expressed as means ± SEM for triplicate samples for all conditions tested. These results are representative of 3 experiments with similar results. *<i>p</i><0.05 compared to uninfected cells (ANOVA).</p

<i>Shigella</i> vaccine strains have attenuated effects on Caco2 monolayer barrier function, except at the highest inoculum.

<p><b>A, C, E.</b> TEER of Caco2 cells infected with wild-type <i>S</i>. <i>flexneri</i> 2a and vaccine candidate <i>S. flexneri</i> 2a CVD 1208S applied apically at inocula of 10⁵, 10⁶ and 10⁷ CFU. <b>B, D, F.</b> TEER of Caco2 cells infected with wild-type <i>S</i>. <i>dysenteriae</i> 1 and attenuated strain <i>S. dysenteriae</i> 1 CVD 1256 applied apically at inocula of 10⁵, 10⁶ and 10⁷ CFU. Data are expressed as means ± SEM for triplicate samples for all conditions tested. * and ∧ indicate statistically significant differences vs. t = 0 and vs. wild-type <i>Shigella</i> at the same inoculum and time point, respectively. # denotes statistically significant differences vs. the uninfected control at the same time point; <i>p</i><0.05 (ANOVA). Statistical analysis results are shown only for the attenuated strains. Please, refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085211#pone-0085211-g001" target="_blank">Figure 1</a> for wild-type strains. These results are representative of 3 experiments with similar results.</p

Wild type <i>Shigella</i> and attenuated strains induce Caco2 cells polarized secretion of IL-8.

<p>IL-8 secreted by Caco-2 cells infected with wild-type live, filtered and heat-killed <i>S. flexneri</i> 2a or vaccine candidate <i>S. flexneri</i> 2a CVD 1208S, applied at different inocula. <b>B</b>. IL-8 released by Caco2 cells infected with wild-type live, filtered and heat-killed <i>S. dysenteriae</i> 1 or <i>S. dysenteriae</i> 1 CVD 1256 applied at different inocula. Statistical comparisons are between infected and uninfected cells (<b>#</b>) or between the attenuated and the wild-type strains at the same inoculum (*). ***<i>p</i><0.001, **<i>p</i><0.01, * and #<i>p</i><0.05 (ANOVA).</p

Tight-junction organization is disrupted following exposure to wild type <i>Shigella</i> but not to vaccine candidates.

<p><b>A, D, G.</b> Uninfected monolayer immunostained for ZO-1, Claudin-1 or Occludin, respectively. <b>B, C.</b> Caco2 cells infected with wild-type <i>S. flexneri</i> 2a (B) or CVD 1208S (C) at 10⁶ CFU, stained for ZO-1 (1∶100). <b>E, F.</b> Caco-2 monolayers infected with wild-type <i>S. flexneri</i> 2a (E) or CVD 1208S (F) at 10⁶ CFU and stained for Claudin-1 (1∶1000). <b>H, I.</b> Monolayers infected with wild-type <i>S. flexneri</i> 2a (H) or CVD 1208S (I) at 10⁶ CFU and stained for Occludin (1∶100). <b>J, K, L.</b> Caco-2 cells treated with <i>S. dysenteriae</i> 1 filtered supernatant (J), live wild-type (K) or attenuated CVD 1256 (L) <i>S. dysenteriae</i> 1 strains applied at 10⁵ CFU and stained for ZO-1 (1∶100). Nuclei are stained with DAPI (blu). Bar 25 µm.</p

Lamina propria architecture in A-CD patients.

<p>Small intestinal biopsies were immunostained with alpha-SMA (green), desmin (red) and DAPI (blue) to visualize SMC, SMC precursor, ISEMF and nuclei. (<i>A</i>) Arrowheads indicate SMC fibers positive for alpha-SMA and desmin in HC (top) and A-CD (bottom) villus core. Arrow indicates desmin-only positive round-shaped cell representative of SMC precursors. (B) Asterisks indicate ISEMFs in the crypt region of HC (top) and A-CD (bottom). Arrowheads indicate SMCs along the crypt in A-CD. Arrow indicates desmin-only positive round-shaped cell representative of SMC precursors. (<i>C</i>) Lower magnification of exemplary intestinal biopsy of HC and A-CD. Normal villus architecture is visible in HC with well defined SMC fibers along the villus axis. In A-CD, swelling of the mucosae is accompanied by crypt hypertrophy and loss of villus structure. ISEMFs are indicated by arrow. <i>Scale bar</i> = 50μm.</p

SOX9 in HC and A-CD.

<p>(A) IHC of SOX9 was performed on (N = 6) HC and (N = 6) A-CD to evaluate the WNT responding cell compartment. (B) The length of the SOX9^<b>+</b> cell domain was measured in N = 3 samples per group using NIS-Elements software and is represented in μm. An average of eight crypts per sample was scored. <i>Scale bar</i> = 100μm. Data are represented as a mean ± SD. (***) <i>P <</i> .<i>001</i>.</p