Tenfold magnification of Masson’s trichome staining of acute, 3 weeks, and 3 months follow-up histology.

<p>Acute (A), 3 weeks (B), and 3 months (C) histology results (10x magnification) showing the border zone of lesions in treated vessels. Sections were stained with MST staining. The arrows indicate the border zone. (A) Border zone of lesion immediately after denervation. The border zone is characterized by a red to pink color transition. At the border zone cell depletion is present (white holes). (B) Border zone of lesion three weeks after denervation. The border zone is characterized by a red to light green color transition. At the site of de border zone inflammatory cells are present (tiny dark blue/black spots). (C) Border zone of lesion 3 months after denervation. The border zone is characterized by blue to red color transition. At the border zone increased collagen deposition is present (blue fibers) and the collagen fibers are intertwined the adjacent healthy muscle tissue of the media (red).</p

Immunohistochemical staining of nerves of treated arteries.

<p>Acute (A-D), 3 weeks (E-L) and 3 months (M-P) histology and immunohistochemical staining results of nerves within the lesion area of treated vessels. The rectangular boxes are a 10x magnification of the affected nerves. a-m shows a 2x magnification and n-p a 5x magnification. Immediately after denervation no signs of nerve damage and a scattered presence of inflammatory cells were observed (A). S100 (B), PGP9.5 (C), and TH (D) showed similar staining patterns and the staining intensity was similar to controls. 3 weeks after denervation neural degeneration and inflammation of nerves and perineural tissue (F) was observed. S100 staining intensity of affected nerves was similar to control (F,J) PGP9.5 staining was slightly lower in intensity (G,K)and TH (H,L)staining was weak or absent compared to control. Scattered presence of S100 (j)and PGP9.5 (K) positive neuron cell bodies was observed around a part of the affected nerves. 3 months after denervation the majority of nerves were embedded in thick sheets of fibrotic tissue(M) and there was scattered presence of inflammatory cells (M). S100, PGP9.5, and TH showed similar staining patterns and the staining intensity was similar to controls. Around affected nerves small S100 (N), PGP9.5 (O) and TH (P) positive nerve bundles were present and they were embedded in thick sheet of fibrotic tissue (M).</p

Definitions of the parameters used.

<p>APV indicates average peak flow velocity; Paorta, aortic pressure; b, baseline; h, hyperemia</p><p>Definitions of the parameters used.</p

Nerve damage outside the lesion area.

<p>3 weeks histology results showing a treated vessel with nerve damage outside the lesion area. A 20 x magnification (a-e) zooms in on the affected nerve that is indicated with an arrow in picture 1–6. Serial sections were stained with HE, MST, α-SMA, S100, PGP9.5 and TH. The perineurial tissue and nerves located at the opposite site of the lesion were affected by an extensive inflammatory response (1,A and 2,B), increased proliferation of myofibroblasts (3,C), a reduction in neural tissue (4,D;5,E) and loss of neurotransmitter production of the affected nerves (6).</p

Relation between minor allele frequencies and levels of plasma TG.

<p>Rs964184 minor allele frequency (95% confidence interval) at increasing levels of TG. N indicates the number of patients with a plasma TG level in this category.</p

Histology and imunnostaining of control arteries.

<p>Histology results and immunostaining of a control vessel. Serial sections were stained for HE, MST, α-SMA, S100, PGP9.5 and TH. Sections show no signs of vessel damage (HE, MST), they have minimal inflammation (HE, MST) and have no increased areas of α-SMA at the media and no staining outside the media, except the arterioles. The nerves show similar staining patterns for structural (S100, PGP9.5) and functional (TH) nerve components.</p

Relation between rs964184 (per minor allele) and lipid levels.

<p>Linear regression adjusted for age and sex.</p><p>*Additionally adjusted for lipid-lowering medication.</p

Acute, 3 weeks, and 3 months follow-up histology results.

<p>Acute (A), 3 weeks (B) and 3 months (C) histology results (2x magnification) showing treated vessels with lesions and control vessels. Serial sections were stained with Haematoxylin Eosin (HE), Masson’s trichome staining (MST) and alpha-smooth muscle actin (α-SMA immunostaining). * = lesion area. (A). HE and MST staining showing a treated vessel with two lesions immediately after denervation. The lesions have a pale color and the media is most affected. MST shows no increased collagen deposition at the site of the lesion (no increased presence of blue fibers). α-SMA staining shows a diffuse increased medial staining at the site of the lesion in treated vessels. (B) HE and MST staining at three weeks follow-up. The media and adventitia of treated vessels are most affected by denervation. MST staining shows increased medial collagen deposition (blue fibers). α-SMA staining shows increased medial, adventitial and perineural staining at the site of the lesion (dark brown). (C) HE and MST staining showing a treated vessel with two lesions 3 months after denervation. MST staining shows transmural collagen deposition at the site of the lesion and the adventitia is most affected. α-SMA staining shows a slightly increased medial staining (dark brown) at the site of the lesion.</p

Overall change in renal hemodynamic parameters directly after renal denervation and after termination.

<p>Continuous variables are displayed as mean ± SD or median (range). The Students T-test or Wilcoxon signed rank test were used for paired samples analysis. APV indicates average peak flow velocity, MR indicates microvascular resistance.</p><p>*Directly after measures compared to baseline measures,</p><p>‡Termination measures compared to baseline measures.</p><p>Overall change in renal hemodynamic parameters directly after renal denervation and after termination.</p

Association between body mass index and plasma log(TG), stratified by rs964184 genotype.

<p>Black line: CC genotype, red line: CG genotype, blue line: GG genotype. Dotted lines indicate 95% confidence interval.</p