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

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

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

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

Female infected versus Control.

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

Proteomic Analysis of Urine Exosomes Reveals Renal Tubule Response to Leptospiral Colonization in Experimentally Infected Rats

<div><p>Background</p><p>Infectious <i>Leptospira</i> colonize the kidneys of reservoir (e.g. rats) and accidental hosts such as humans. The renal response to persistent leptospiral colonization, as measured by urinary protein biosignatures, has not been systematically studied. Urinary exosomes--bioactive membrane-bound nanovesicles--contain cell-state specific cargo that additively reflect formation all along the nephron. We hypothesized that <i>Leptospira</i>-infection will alter the content of urine exosomes, and further, that these <i>Leptospira</i>-induced alterations will hold clues to unravel novel pathways related to bacterial-host interactions.</p><p>Methodology/Principal findings</p><p>Exosome protein content from 24 hour urine samples of <i>Leptospira</i>-infected rats was compared with that of uninfected rats using SDS-PAGE and liquid chromatography/tandem mass spectrometry (LC-MS/MS). Statistical models were used to identify significantly dysregulated proteins in <i>Leptospira</i>-infected and uninfected rat urine exosomes. In all, 842 proteins were identified by LC-MS/MS proteomics of total rat urine and 204 proteins associated specifically with exosomes. Multivariate analysis showed that 25 proteins significantly discriminated between uninfected control and infected rats. Alanyl (membrane) aminopeptidase, also known as CD13 topped this list with the highest score, a finding we validated by Western immunoblotting. Whole urine analysis showed Tamm-Horsfall protein level reduction in the infected rat urine. Total urine and exosome proteins were significantly different in male vs. female infected rats.</p><p>Conclusions</p><p>We identified exosome-associated renal tubule-specific responses to <i>Leptospira</i> infection in a rat chronic colonization model. Quantitative differences in infected male and female rat urine exosome proteins vs. uninfected controls suggest that urine exosome analysis identifies important differences in kidney function that may be of clinical and pathological significance.</p></div