Proinflammatory cytokines, adhesion molecules and c-IMT measurements.

<p>A) Differences in the gene expression of proinflammatory markers in the artery wall according to c-IMT tertiles. B) Differences in the quantification of proinflammatory proteins by c-IMT tertiles. *ANOVA test for VCAM-1, <i>P</i> = 0.003; Bonferroni procedure, T3 vs. T1, p = 0.003; T3 vs T2, <i>P</i> = 0.076.</p

Relationship between VCAM protein levels and overall mortality by bivariate and multivariate Cox regression analysis

<p>Abbreviations: c-IMT, carotid intima-media thickness; KT, kidney transplantation; VC, vascular calcifications; NODAT, new onset diabetes after transplantation; PD, peritoneal dialysis; T-cholesterol, total cholesterol. Group II, patients who showed an increase to the highest tertile or who maintained both values within the highest tertile; Group I, patients who showed a reduction to a lower tertile or who maintained both values within the lower or the middle tertile.</p><p>*Because the number of events was few, this analysis was performed entering risk factors two by two.</p><p>Relationship between VCAM protein levels and overall mortality by bivariate and multivariate Cox regression analysis</p

Artery Wall Assessment Helps Predict Kidney Transplant Outcome

<div><p>Background</p><p>Kidney transplant recipients have high cardiovascular risk, and vascular inflammation may play an important role. We explored whether the inflammatory state in the vessel wall was related to carotid intima-media thickness (c-IMT) and patient survival following kidney transplantation.</p><p>Methods</p><p>In this prospective observational cohort study we measured c-IMT and expression of proinflammatory cytokines and adhesion molecules in the inferior epigastric artery in 115 kidney transplant candidates. Another c-IMT measurement was done 1-year post-transplantation in 107. By stepwise multiple regression analysis we explored factors associated with baseline c-IMT and their changes over time. Multivariate Cox regression analysis was constructed to identify risk factors for mortality.</p><p>Results</p><p>A worse cardiovascular profile (older age, smoker, diabetic, carotid plaque, systolic blood pressure and vascular calcification) and higher VCAM-1 levels were found in patients in the highest baseline c-IMT tertile, who also had a worse survival. Factors independently related to baseline c-IMT were age (β=0.369, <i>P</i><0.0001), fasting glucose (β=0.168, <i>P</i>=0.045), smoking (β=0.228, <i>P</i>=0.003) and VCAM-1 levels (β=0.244, <i>P</i>=0.002). Independent factors associated with c-IMT measurement 1-year post-transplantation were baseline c-IMT (β=-0.677, <i>P</i><0.0001), post-transplant diabetes (β=0.225, <i>P</i>=0.003) and triglycerides (β=0.302, <i>P</i>=0.023). Vascular VCAM-1 levels were associated with increased risk of mortality in bivariate and multivariate Cox regression. Notably, nearly 50% of patients showed an increase or maintenance of high c-IMT 1 year post-transplantation and these patients experienced a higher mortality (13 versus 3.5%; <i>P</i>=0.021).</p><p>Conclusion</p><p>A worse cardiovascular profile and a higher vascular VCAM-1 protein levels at time of KT are related to subclinical atheromatosis. This could lead to a higher post-transplant mortality. Pre-transplant c IMT, post-transplant diabetes and triglycerides at 1-year post-transplantation may condition a high c-IMT measurement post-transplantation, which may decrease patient survival.</p></div

Functional characterization of WT and p.H83R.

<p>A and B) Enzyme activity measurements for WT (A) and p.H83R (B) at different L-Ala concentrations in the presence of 0.25 mM (diamonds), 0.5 mM (down triangles), 1 mM (up triangles) and 2 mM (circles) glyoxylate. Data in panel A are means±s.d from four independent measurements while data in panel B are means from two independent measurements. Lines are best fits for the different glyoxylate concentrations obtained from global fittings using a double-displacement mechanism. C and D) Absorption (C) and circular dichroism (D) spectra for holo-WT (black) and holo-p.H83R (grey) acquired upon incubation for at least 10 min in the absence (continuous lines; PLP bound) or presence (dashed lines; PMP bound) of 500 mM L-alanine.</p

Functional properties and hydrodynamic diameter of AGT variants.

a<p>Enzyme kinetic parameters were obtained from global fittings of 2–4 independent experimental series using a double-displacement mechanism;</p>b<p><i>K</i>_d values were estimated from single titrations except for AGT WT protein where <i>K</i>_d value is the mean±s.d. from three independent titrations. Data in parenthesis are <i>K</i>_d estimates obtained from the kinetic binding experiments (<i>K</i>_d = <i>k</i>_off/<i>k</i>_on).</p>c<p>Hydrodynamic diameter of holo-AGT variants determined by dynamic light scattering (DLS). Data are mean±s.d. of 3–9 independent measurements.</p

Clinical characteristics of patients after stratification according to c-IMT measurement variation after second carotid echographic study.

<p>Abbreviations: c-IMT, carotid intima-media thickness; CVD, cardiovascular disease; KT, kidney transplantation; ACEI/ARA, angiotensin-converting enzyme inhibitor/angiotensin receptor antagonist; NODAT, new onset diabetes after transplantation.</p><p>Clinical characteristics of patients after stratification according to c-IMT measurement variation after second carotid echographic study.</p

Baseline clinical and pathological data of the artery wall in kidney transplant patients according to carotid intima-media thickness tertiles.

<p>Abbreviations: c-IMT, carotid intima-media thickness; CKD, chronic kidney disease; ACEI/ARA, angiotensin-converting enzyme inhibitor/angiotensin receptor antagonist; IEA, inferior epigastric artery.</p><p>^a<i>P</i><0.0001 vs. T2 and T1;</p><p>^b<i>P</i><0.0001 vs. T1;</p><p>^c<i>P</i> = 0.033 vs T2;</p><p>^d<i>P</i> = 0.012 vs. T1;</p><p>^e<i>P</i> = 0.006 vs. T1;</p><p>^f<i>P</i> = 0.023 vs. T1;</p><p>^g<i>P</i> = 0.001</p><p>Baseline clinical and pathological data of the artery wall in kidney transplant patients according to carotid intima-media thickness tertiles.</p

Structure-energetics relationships for thermal denaturion of holo- and apo-AGT enzymes.

<p>A) Temperature dependence of denaturation enthalpies (Δ<i>H</i>) for holo- (closed symbols) and apo-(open symbols) proteins. The linear fit provides the value of ΔC_p ( = 10.2±0.6 kcal.mol⁻¹). B) Activation energy (<i>E</i>_a) plotted vs. the <i>T</i>_m for holo- (closed symbols) and apo-(open symbols) AGT enzymes. C and D) changes in activation enthalpic and entropic contributions to AGT kinetic stability as a function of changes in activation free energies for holo-(C) and apo-(D) AGT enzymes. Lines in C and D are meant to guide the eye and have no theoretical meaning.</p

Interaction of AGT variants with Hsc70 chaperones in a cell-free system.

<p>Representative autoradiograms of AGT proteins labeled with ³⁵S-Met are shown for several AGT variants (E: total AGT synthesized in extracts; I: AGT immunoprecipitated using anti-Hsc70 antibodies; note that 1 µl of the TnT reaction was loaded in E lanes, while the protein immunoprecipitated from 6 µl TnT lysate was loaded in I lanes). Data in the lower panel are expressed as percentage of immunoprecipitated AGT compared to the total AGT synthesized, and are mean±s.d. from three independent experiments.</p

Kaplan-Meier curves according to variation patterns between the c-IMT tertiles at both time periods.

<p>Solid line indicates the “decrease or stable low-middle” group and dotted line the “increase or stable high” group (log-rank analysis 5.4; <i>P</i> = 0.021).</p