Correlation of coarse particulate matter (PM_10-2.5) pollution with Emergency Department daily admissions for venous thromboembolism by unadjusted and multiple adjusted Poisson regression.

†<p>: adjusted for atmospheric variables (i.e. mean temperature, humidity, and air pressure).</p>‡<p>: adjusted for atmospheric variables and PM_2.5.</p

Coarse particulate matter (PM_10-2.5) and daily admissions to the Emergency Department for venous thromboembolism (VTE).

<p>Data are presented as mean level of PM_10-2.5 concentration (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034831#pone-0034831-g002" target="_blank">Figure 2A</a>) and prevalence of days with high PM_10-2.5 concentration, defined as higher than the 75^th percentile – 19 mcg/m³ (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034831#pone-0034831-g002" target="_blank">Figure 2B</a>), according to the number of daily admissions to the Emergency Department for VTE.</p

Seasonal trend of total, fine, and coarse particulate matter (PM₁₀, PM_2.5, and PM_10-2.5) concentrations during the study period.

<p>In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034831#pone-0034831-g001" target="_blank">Figure 1A</a>, PM₁₀ levels are represented by the orange line and the area under the curve is divided in the 2 components, PM_2.5 represented by the green area and PM_10-2.5 represented by the ochre yellow area. In <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034831#pone-0034831-g001" target="_blank">Figure 1B</a>, the seasonal trend of PM_10-2.5 concentrations is separately represented and related with data of daily admissions for venous thromboembolism (VTE). The dashed line represents the PM_10-2.5 75^th percentile, at 19 µg/m³.</p

Association between daily hospital referral for venous thromboembolism (VTE) and coarse particulate matter (PM_10-2.5) concentration at different time-lags.

<p>The association was estimated by using distributed lag non-linear models with VTE risk as outcome and time-lags expressed as the number of previous days. Only the current-day (lag 0) PM_2.5-10 levels presented a significant association with VTE risk.</p

Pearson's correlations of particulate matter (PM) concentrations, considered as a whole (PM₁₀) or subdivided in the finest (PM_2.5) and the coarse component (PM_10-2.5), with activated partial thromboplastin time (aPTT) and prothrombin time (PT).^*

*<p>: This analysis was performed in a subgroup of patients admitted to Emergency Department for mild respiratory symptoms, without active thrombosis and not taking anticoagulant therapy, for whom data about coagulation times were available (n = 102).</p

Effect of the use of an internal standard on the hepcidin-25 concentrations in urine and serum of selected clinical populations.

<p>Hepc25 concentrations were calculated in nM based on the known concentration of spiked hepc24 in serum (A) and urine (B) samples. For serum, hepc24 intensities were corrected for the background intensity of unspiked samples (hepc24-bl). Note that for both urine and serum specimens the LLOD depends on the individual sample matrix and therefore varies between samples. The LLOD was determined in the 25 human serum and urine samples by using the background intensities at m/z 2400, 2515, 2846 for serum and at m/z 2299, 2510, 2910, for urine samples, respectively. The detection limit was defined as the mean+2 SD of these measurements and found be 2.0 peak intensity for serum and 1.76 peak intensity for urine. The lower level of detection (LLOD) in nM of each individual sample was determined by incorporating the sample specific hepc24 peak intensity value and these mean LLOD values in peak int for hepc25 peak at 2789 m/z in the formulas 1 and 2 for urine and serum (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002706#s4" target="_blank">Material and Methods</a> section). Ctrl, control; LPS, volunteers injected with polysaccharide (6 h after injection); IDA, iron deficiency anemia; TM, thalassemia major in various stages of disease; HH, C282Y homozygous hereditary hemochromatosis patients of various stages of disease; ◊, hepcidin concentration; Ж, sample specific LLOD, the hepcidin concentration of the sample is then</p

Fractional excretion of hepcidin.

<p>The fractional excretion (FE) is calculated by: urine hepcidin (nM)×serum creat (µM)×100/serum hepcidin (nM)×urine creat (mM)×1000. n, the number of pairs for which all data needed to calculate the FE were available. Ctrl, controls; LPS, volunteers injected with polysaccharide (6 h after injection); TM, thalassemia major in various stages of disease; HH, C282Y homozygous hereditary hemochromatosis patients at various stages of disease; tubular reabsorbtion (%) = 100- FE (%).</p

Characteristics of healthy control subjects.

<p>Data are means with 95% C.I. and p-values of males <i>vs</i> females by t-test. TS, transferrin saturation; sTfR, soluble transferrin receptor.</p><p>Body iron stores were calculated on the basis of the logarithm of the concentrations in micrograms of serum transferrin receptor/serum ferritin (TfR/ferritin ratio) and expressed as milligram per kilogram body weight, as follows: body iron (mg/kg) = −[log(TfR/Ferritin ratio) −2.8229]/0.1207 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002706#pone.0002706-Cook1" target="_blank">[30]</a>. Positive values represent the iron surplus in stores, while negative values represent iron deficit in tissues.</p

Correlation between serum hepcidin and ferritin.

<p>Serum hepcidin levels in 23 healthy volunteers (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002706#pone-0002706-t002" target="_blank">Table 2</a>), as determined by our updated MS method, were correlated with their ferritin levels. Values were log transformed prior to correlation analysis. Pearson correlation: 0.6804 (p = 0.0004).</p

SELDI-TOF MS profiles obtained in the different experiments.

<p>SELDI-TOF MS profiles of (A) hepidin-24 spiked urine sample showing next to the expected hepcidin forms also methionine oxidized (Ox) forms of Hepc24 and Hepc25; (B and C) different patient sera and urines, respectively, spiked with Hepc24 (5 nM into urines and 10 nM into sera). Note, the influence of the serum and urine matrices on the peak height of the Hepc24 spiked to patient samples; (D and E), blank serum and urine samples spiked with both Hepc24 and Hepc25 (7.5 nM of both hepcidin forms into urine and 10 nM into serum). Note that the method appears to be more sensitive for Hepc25 than for Hepc24, with an average peak intensity ratio Hepc24/Hepc25 of 0.693. This is probably due to the absence of a negatively charged aspartic acid residue in Hepc24, which negatively affects its binding on the IMAC-Cu²⁺ protein chip surface. The hepcidin isoforms Hepc20, Hepc22 (only in urine), Hepc24 (synthetic analogue) and Hepc25 are indicated by arrows.</p