Proteomic analyses of Urine Exosomes reveal New Biomarkers of Diabetes in Pregnancy.

ObjectiveTo evaluate 24 hour urine exosome protein content changes among pregnant US subjects with diabetes and obesity during early pregnancy.MethodsThe exosome proteome content from 24 hour urine samples of pregnant subjects with gestational diabetes mellitus (GDM, N=8) and pre-gestational Type 2 diabetes (PGD, N = 10) were compared with control samples (CTRL, N = 10) obtained at week 20 of pregnancy. Differences in exosome protein load between groups was identified by liquid chromatography/mass spectrometry, analyzed by linear regression in negative binomial distribution, visualized in MetaboAnalyst (version 3.0), and validated by western immunoblotting.ResultsAt the 20th week of pregnancy, we identified 646, 734 and 856 proteins in exosomes from 24 hour urine samples of patients from the CTRL, GDM and PGD groups, respectively. S100 calcium binding protein A9, damage associated molecular pattern (DAMP) signal, was found to be significantly increased in both GDM and PGD subjects. In GDM subjects the peptide counts for S100A9 protein independently correlated with maternal obesity and macrosomia of the newborn infants. Early to late pregnancy developmental changes in the GDM group were shown to utilize pathways and protein expression levels differently from those in PGD or CTRL groups.ConclusionsUrinary exosome proteomic analysis non-invasively provides insights into maternal changes during diabetic pregnancy. Exosome biomarkers early in pregnancy can be potentially used to better understand pathophysiologic mechanisms of diabetes at a cellular level, and to distinguish between gestational and pre-gestational diabetes at the pathway level. This information can aid intervention efforts to improve pregnancy outcomes in women with diabetes

Stress errors in polysyllabic adjective words by the 6th semester students of English Letters Department in Sanata Dharma University

<p>Although clear separation between control and infected rat urine exosomes is seen, a fraction of overlap between the infected male and infected female rat urine exosome proteins is observed.</p

Proteomic analyses of Urine Exosomes reveal New Biomarkers of Diabetes in Pregnancy.

ObjectiveTo evaluate 24 hour urine exosome protein content changes among pregnant US subjects with diabetes and obesity during early pregnancy.MethodsThe exosome proteome content from 24 hour urine samples of pregnant subjects with gestational diabetes mellitus (GDM, N=8) and pre-gestational Type 2 diabetes (PGD, N = 10) were compared with control samples (CTRL, N = 10) obtained at week 20 of pregnancy. Differences in exosome protein load between groups was identified by liquid chromatography/mass spectrometry, analyzed by linear regression in negative binomial distribution, visualized in MetaboAnalyst (version 3.0), and validated by western immunoblotting.ResultsAt the 20th week of pregnancy, we identified 646, 734 and 856 proteins in exosomes from 24 hour urine samples of patients from the CTRL, GDM and PGD groups, respectively. S100 calcium binding protein A9, damage associated molecular pattern (DAMP) signal, was found to be significantly increased in both GDM and PGD subjects. In GDM subjects the peptide counts for S100A9 protein independently correlated with maternal obesity and macrosomia of the newborn infants. Early to late pregnancy developmental changes in the GDM group were shown to utilize pathways and protein expression levels differently from those in PGD or CTRL groups.ConclusionsUrinary exosome proteomic analysis non-invasively provides insights into maternal changes during diabetic pregnancy. Exosome biomarkers early in pregnancy can be potentially used to better understand pathophysiologic mechanisms of diabetes at a cellular level, and to distinguish between gestational and pre-gestational diabetes at the pathway level. This information can aid intervention efforts to improve pregnancy outcomes in women with diabetes

A. Immunoblotting of rat urine exosome for CD13 protein.

<p>Lanes 1–3: control; lanes 4–6: lepto infected female; lanes 7–9: infected male. B. Quantification of CD13 from immunoblots in A (control, n = 3; infected, n = 6). Data are means ± SEM. p<0.05 rat urine exosome CD13 <i>Leptospira</i>- infected versus uninfected control.</p

Top discriminators between control, infected male & infected female rat urine exosome proteins.

<p>Top discriminators between control, infected male & infected female rat urine exosome proteins.</p

Proteins significantly dysregulated between exosomes from urines of control rats and infected rats.

<p>Proteins significantly dysregulated between exosomes from urines of control rats and infected rats.</p

A. Immunoblotting of rat urines for Tamm-Horsfall Protein (THP).

<p>Left panel Lanes (1–3): Control Uninfected Rat urines; Right panel Lanes (1–6): <i>Leptospira</i>-Infected Rat urines. B. Quantification of THP from immunoblots in A (control, n = 3; infected, n = 6). Data are means ± SEM. p<0.05 rat urine THP, <i>Leptospira</i>-infected versus uninfected control.</p

Variable importance in projection (VIP) plot: important features (analyzed serum free amino acids) identified by PLS-DA in a descending order of importance.

<p>The graph represents relative contribution of proteins to the variance between the <i>Leptospira</i>-infected and uninfected control rat urine exosomes. High value of VIP score indicates great contribution of the proteins to the group separation. The green and red boxes on the right indicate whether the protein concentration is increased (green) or decreased (red) in the exosome of the infected rat urine vs. uninfected rat urine samples. For higher n value, a VIP score of 1.5 is considered to enable discrimination between 2 phenotypes. Even with the low n (= 3) per group that is employed in this study, the VIP score of the top 3 proteins is higher than 3, increasing the confidence. Alanyl (membrane) aminopeptidase, also called CD13 is the top protein with a VIP score of 5.72.</p