CMV infection of placental villous explants results in upregulation of MCP-1 and TNF-α expression.

<p>(A) MCP-1 and TNF-α expression in mock-infected, AD169-infected (0.2 pfu/ml) and Merlin-infected (0.02 pfu/ml) villous explants. Data presented as box plots with median value, Q1, Q3 and range. Significant differences between groups were determined with the Mann-Whitney U test and threshold for significance adjusted to account for multiple comparisons using Bonferroni’s correction; *<i>p</i><0.025, **<i>p</i><0.005 and ***p<0.0005. (B) Correlation between degree of CMV placental explant infection with corresponding MCP-1 and TNF-α expression. MCP-1 and TNF-α protein expression was plotted against the number of cells expressing CMV immediate early/early (IE/E) protein per mm² of AD169 and Merlin infected villous explant tissue. Significant correlations (denoted by *) were determined by two-tailed nonparametric spearmans correlation with trend plotted as straight lines.</p

MCP-1 and TNF-α expression is elevated in CMV-infected placentae from stillborn babies.

<p>(A) MCP-1 and TNF-α expression in uninfected (CMV−/Other−), other microorganism-infected (CMV−/Other+) and CMV-infected (CMV+/Other−) placentae from stillborn babies. Data presented as box plots with median value, Q1, Q3 and range. Significant differences between groups were determined with the Mann-Whitney U test and threshold for significance adjusted to account for multiple comparisons using Bonferroni’s correction; *<i>p</i><0.0167, **<i>p</i><0.003. (B) Representative images of immunohistochemical localisation of MCP-1 and TNF-α protein (red staining) in uninfected and CMV-infected placental tissue from stillborn babies. In chorionic villi, cytokine localised in syncytiotrophoblast (st) and cytotrophoblast (ct) cells, mesenchymal stromal cells (msc) and endothelial cells of fetal capillaries (ec). Scale bars represent 70 µm.</p

CMV productively infects placental villous explants.

<p>Productive AD169 and Merlin infection of villous explants was determined by staining for CMV immediate early/early (IE/E), early/late (pp65) and late (gB) protein. Representative images are of AD169 and Merlin infected villous explants 12 days post inoculation. Scale bars represent 50 µm.</p

CMV infection is significantly greater in AD169- compared with Merlin-infected placental villous explants.

<p>(A) Representative images of CMV immediate early/early (IE/E) antigen detection in AD169- (left) and Merlin-infected (right) placental villous explants 12 days post inoculation (CK7; Cytokeratin-7). Scale bar represents 70 µm. (B) Number of cells per mm² of villous explant tissue expressing CMV IE/E protein in AD169- compared with Merlin-infected explants. (C) No significant differences in villous explant area (mm²) were observed between the CMV-infected explant groups (<i>p</i> = 0.1). Data presented as box plots with median value, Q1, Q3 and range. Significant differences between groups (denoted as *) were determined using a one-tailed Spearman’s correlation.</p

CMV actively replicates in cytotrophoblast and mesenchymal cells, but not syncytiotrophoblasts, of placental villous explants.

<p>CMV laboratory strain AD169 and wild-type Merlin infected syncytiotrophoblasts (CK7+/VIM−/hPL+), cytotrophoblasts (CK7+/VIM−/hPL−) and mesenchymal cells of the villous stroma (CK7−/VIM+/hPL−) as determined by staining for CMV immediate early/early protein. Active replication (CMV IE/E+, pp65+) was observed in both cytotrophoblasts and mesenchymal cells but not syncytiotrophoblasts (CMV IE/E+, pp65−) (inserts and arrow heads). Representative images are of 4 µm consecutive histological sections of AD169 infected villous explants 12 days post inoculation. No difference in cellular tropism was observed between AD169 and Merlin strains. Scale bars represent 100 µm.</p

Rate of microvascular complications according to insulin delivery: CSII vs MDI.

<p>Rate of microvascular complications according to insulin delivery: CSII vs MDI.</p

Effect of pump therapy compared to multiple daily injections for microvascular complications.

<p>Multivariable analysis (GEE), after inclusion of HbA1c and other confounders: significant reduction for retinopathy after allowing for HbA1c, age and duration; and significant reduction for peripheral nerve abnormalities after allowing for HbA1c, male gender and height SDS.</p

Characteristics and complication rates in adolescents with type 1 diabetes according to treatment.

<p>Characteristics and complication rates in adolescents with type 1 diabetes according to treatment.</p

Combined exposure to low occupational sick score and low hand washing is associated with greater risk of type 1 diabetes onset: Evidence of interaction.

<p>The combined exposure to low occupational sick score (0–2 vs. rest) and low hand washing (never, occasionally) had an odds ratio of 3.86 (95% CI: 2.08, 7.16); among those with a low occupational sick score and high hand washing the odds ratio was 1.74 (95% CI: 0.89, 3.41); among those with low hand washing and a high occupational sick score the odds ratio was 1.67 (95% CI: 0.86, 3.24) compared to the lowest risk category associated with both high occupational sick score and high hand washing (AOR 1.00 (reference)). The Synergy Index is 5.16 (95% CI: 3.61, 7.36) with a Relative Excess Risk due to Interaction of 37.46 (95% CI: 13.96, 60.95) and Attributable Proportion of 0.79 (95% CI: 0.72, 0.86). The AOR_multi is 1.19, P = 0.03. All odds ratio adjusted for age and sex. Thus, the interaction is evident on the additive and multiplicative scale.</p

Higher parent occupational microbial contact is associated with a reduced risk of type 1 diabetes onset: Ten measures and two composite indices.

<p>Higher parent occupational microbial contact is associated with a reduced risk of type 1 diabetes onset: Ten measures and two composite indices.</p