Integrative multiomics analysis of human atherosclerosis reveals a serum response factor-driven network associated with intraplaque hemorrhage.

BACKGROUND: While single-omics analyses on human atherosclerotic plaque have been very useful to map stage- or disease-related differences in expression, they only partly capture the array of changes in this tissue and suffer from scale-intrinsic limitations. In order to better identify processes associated with intraplaque hemorrhage and plaque instability, we therefore combined multiple omics into an integrated model. METHODS: In this study, we compared protein and gene makeup of low- versus high-risk atherosclerotic lesion segments from carotid endarterectomy patients, as judged from the absence or presence of intraplaque hemorrhage, respectively. Transcriptomic, proteomic, and peptidomic data of this plaque cohort were aggregated and analyzed by DIABLO, an integrative multivariate classification and feature selection method. RESULTS: We identified a protein-gene associated multiomics model able to segregate stable, nonhemorrhaged from vulnerable, hemorrhaged lesions at high predictive performance (AUC >0.95). The dominant component of this model correlated with αSMA- PDGFRα+ fibroblast-like cell content (p = 2.4E-05) and Arg1+ macrophage content (p = 2.2E-04) and was driven by serum response factor (SRF), possibly in a megakaryoblastic leukemia-1/2 (MKL1/2) dependent manner. Gene set overrepresentation analysis on the selected key features of this model pointed to a clear cardiovascular disease signature, with overrepresentation of extracellular matrix synthesis and organization, focal adhesion, and cholesterol metabolism terms, suggestive of the model's relevance for the plaque vulnerability. Finally, we were able to corroborate the predictive power of the selected features in several independent mRNA and proteomic plaque cohorts. CONCLUSIONS: In conclusion, our integrative omics study has identified an intraplaque hemorrhage-associated cardiovascular signature that provides excellent stratification of low- from high-risk carotid artery plaques in several independent cohorts. Further study revealed suppression of an SRF-regulated disease network, controlling lesion stability, in vulnerable plaque, which can serve as a scaffold for the design of targeted intervention in plaque destabilization.

Duplex-guided versus Conventional Percutaneous Transluminal Angioplasty of Iliac TASC II A and B Lesion: A Randomized Controlled Trial.

BACKGROUND: Percutaneous transluminal angioplasty (PTA) requires the use of nephrotoxic contrast. Patients with chronic kidney disease are more prone to develop contrast-induced nephropathy after utilization of contrast. Doppler ultrasound (duplex)-guided PTA (DuPTA) is a novel technique and has recently proven to be a successful alternative to conventional PTA in the treatment of iliac stenotic disease, without the use of contrast. In this randomized controlled trial, we evaluated whether DuPTA is as effective as conventional PTA in the treatment of iliac arterial stenotic disease. METHODS: From June 2013 till January 2017, 142 patients with symptomatic peripheral arterial disease (PAD), with significant (>70%) iliac stenotic lesions (both isolated and patients with multilevel disease without treatment of other lesions), were randomized to receive either conventional PTA or DuPTA of the iliac lesion, regardless of renal function. All patients received an ankle-brachial index, arterial mapping with duplex, and magnetic resonance angiography in the pre-operative work-up. Primary end point was procedural success, defined as passing the guidewire through the stenotic lesion and performing a PTA, with or without stenting. Reduction in peak systolic velocity (PSV) of ≥50% after successful PTA was required in the DuPTA group. Angiographic reduction of more than 50% was required in the interventional group. Decrease in PSV was evaluated in both groups 4 weeks post-procedure. RESULTS: Passing of the guidewire through the stenotic lesion was achieved in 96.5% of the DuPTA group and 98.8% of the PTA group (P = 0.34). Although PSV decreased significantly in both groups 4 weeks post-operative, PSV reduction ≥50% was significantly higher in the DuPTA group, respectively 78% vs. 58% in the PTA group (P < 0.01). The utilization of stents was significantly greater in the DuPTA group (52% vs. 18%, P < 0.01). After correction of potential confounders, significant difference in ≥50% PSV reduction remained; technical success did not differ significantly. CONCLUSIONS: DuPTA is a feasible alternative to conventional PTA in the treatment of PAD on the iliac anatomic level. Duplex examination before removal of the guidewire is recommended to evaluate adequate decrease in PSV and identify potential recoil.

Role of compression stockings after endovenous laser therapy for primary varicosis.

OBJECTIVE: The objective of this prospective, randomized trial was to evaluate the effect of compression stockings after endovenous laser therapy (EVLT) for insufficiency of the great saphenous vein. Effect evaluation was focused on differences in postoperative pain within 6 weeks. METHODS: The study randomized 111 patients undergoing EVLT for duplex ultrasound-confirmed great saphenous vein insufficiency (C2-4EPASPR) to receive either 2 weeks of elastic stockings (class II, thigh length) or no elastic stockings after an initial 24-hour period of wearing bandages. Patients registered pain scores (primary outcome) and use of analgesics in a diary on a visual analog scale. At 2 days, 14 days, and 6 weeks after the procedure, leg circumferences, Aberdeen Varicose Vein Questionnaire scores, RAND 36-Item Health Survey scores, patient satisfaction (5-point scale), time before returning to work, and adverse events (secondary outcomes) were evaluated. RESULTS: In the 79 patients (15 men and 64 women; mean age, 52 years) who completed the treatment and follow-up, small but significant differences in pain scores were observed during the first week after laser surgery, with more favorable scores in the group wearing stockings. Patients not wearing stockings used more analgesics than did patients wearing stockings (P < .05). Patients wearing stockings reported a statistically significantly higher score of satisfaction at 2 days (4.44 vs 4.15) and 6 weeks (4.59 vs 4.18), although the absolute differences were small. No significant differences were found regarding time to return to work, Aberdeen Varicose Vein Questionnaire scores, RAND 36-Item Health Survey scores, leg circumference measurements, and risk of complications. CONCLUSIONS: Wearing of postoperative stockings for 2 weeks after an initial 24-hour period of wearing bandages results in a small but significant reduction of postoperative pain and use of analgesics compared with not wearing compression stockings after EVLT for great saphenous vein insufficiency.

Intercellular adhesion molecule-1 and gelatinase expression in human peritoneal mesothelial cells during propagation in culture.

Mesothelial cells are involved in a variety of biological processes, which include the formation of peritoneal adhesions. The cultures of human peritoneal mesothelial cells comprise an important tool to investigate the behavior and functions of mesothelial cells. Very little is known about the differences among mesothelial cells isolated from different sources and about the changes in specific functions as caused by cell propagation in vitro or that result from storage of cells at low temperatures. This study aims to characterize 2 particular cellular activities relevant for tissue repair, which include the expression of intercellular adhesion molecule-1 (ICAM-1) and the gelatinase activity; in addition, this study will assess the effect of hyaluronan, which is an antiadhesive agent, on these cellular activities. Viable cell lines were established from both omentum and peritoneal lavage fluid from 7 patients. Both ICAM-1 expression, which was measured by enzyme-linked immunosorbent assay (ELISA), and matrix metalloproteinase (MMP) bioactivity, which was measured by zymography, were measured in the 2nd and 4th passage; the latter also was measured after freezing and storing of cells in liquid nitrogen. The effects of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), phorbol myristate acetate (PMA), and hyaluronan were analyzed. ICAM-1 was constitutively expressed and stimulated by IL-1beta, TNF-alpha, and PMA. All cell lines produced both MMP-2 and MMP-9. Only the latter activity was affected by TNF-alpha and, especially so, PMA. Differences were found between the 2nd and 4th passage, as well as between cells of different lineage, mostly so if the relative stimulation by the various agents was compared. The addition of sodium hyaluronate either to control cultures or to cultures together with any of the 3 stimuli examined did not significantly change either ICAM-1 expression or gelatinase activity. The freezing and storage of cells did not affect their functions. Both the human omentum and peritoneal lavage fluid are good sources to establish mesothelial cell lines, which can be propagated also after freezing without qualitative changes in their ability to express ICAM-1 and produce the gelatinases. For omental cells, a differential effect of stimulation occurs depending on whether the cells have been passaged 2 or 4 times. The presence of hyaluronan did not affect the expression of ICAM-1 or the gelatinases.

Tc-99m-PEG-liposomes target both adhesions and abscesses and their reduction by hyaluronate in rats with fecal peritonitis.

BACKGROUND: Abdominal adhesions and abscesses are a major source of morbidity and mortality after abdominal surgery and peritonitis. Adhesions are hard to detect with standard imaging techniques. Liposomes, coated with polyethylene glycol (PEG), represent an agent developed for infection imaging. This study investigated the capacity of 99mTc-PEG-liposomes to localize early adhesion formation after peritonitis. Additionally, the value of 99mTc-PEG-liposomes for therapy evaluation of hyaluronan solution, which reduces adhesion and abscess formation in experimental peritonitis, was assessed. METHODS: In 24 rats, a bacterial peritonitis was induced by performing a cecal ligation and puncture procedure. The animals were treated with sodium chloride solution or 0.4% hyaluronan solution intra-abdominally. One week later, scintigraphy was performed using 99mTc-PEG-liposomes, and abnormal focal uptake in the abdomen was scored. Thereafter, autopsy was performed and adhesions and abscesses were scored. RESULTS: A significant correlation was found between the total adhesion score and the scintigraphic score (P < 0.01, r = 0.65). Treatment with hyaluronan significantly reduced the total adhesion score (P = 0.01). The size of abscesses significantly correlated with the scintigraphic score (P < 0.01, r = 0.65). Treatment with hyaluronan reduced the size of abscesses (P < 0.05). CONCLUSION: 99mTc-PEG-liposomes are able to detect early adhesions and abscesses and may be used for therapy evaluation of agents that reduce adhesions and abscesses.

Hyaluronan-based antiadhesive agents in abdominal surgery: applications, results, and mechanisms of action.

Postsurgical intra-abdominal adhesions cause significant morbidity and mortality, with small bowel obstruction being the most common complication. The urge to prevent adhesion formation has resulted in multiple experimental and clinical trials and the development of numerous antiadhesive agents. Through the years, hyaluronan-based antiadhesives have proved to be successful in the reduction of adhesion formation. Despite the obvious effectiveness of hyaluronan, there is still much debate on its clinical use and mechanisms of action. Various hyaluronan-containing products have been introduced and withdrawn from the market. The application of hyaluronan in combination with meshes for hernia repair appears to be a promising concept. Not all different applications of hyaluronan are well known and its use in patients with a malignancy or abdominal infection remains controversial. Here an overview is given on the effects of hyaluronan-based antiadhesive agents in abdominal surgery, its use in infectious conditions, and its oncologic repercussions. The most important mechanism of action appears to be the mechanical separation of damaged peritoneal surfaces. However, the biological effects of hyaluronan, such as modulation of cell proliferation and peritoneal biology, might also be of influence.

Auto-cross-linked hyaluronic acid gel does not reduce intra-abdominal adhesions or abscess formation in a rat model of peritonitis.

BACKGROUND: Prevention of adhesion and abscess formation would decrease mortality and morbidity after peritonitis. In this study the effect of a new anti-adhesive, auto-cross-linked hyaluronic acid polysaccharide (ACP) gel, on adhesion and abscess formation was studied in a rat peritonitis model. MATERIALS AND METHODS: In experiment 1, bacterial peritonitis was induced in 24 Wistar rats, using a cecal ligation and puncture model. Animals were randomized to receive 4 mL ACP gel (4%) or 4 mL phosphate buffered saline (PBS). After 2 weeks animals were killed and adhesions and abscesses were scored. In experiment 2, 72 rats underwent the same procedure but were randomized to receive 2 mL ACP gel, 4 mL ACP gel, or 4 mL PBS. After 1 and 3 weeks, respectively, half of the animals in each group were killed and adhesions and abscesses were scored. RESULTS: The median total adhesion score was 12 (range, 3-20) in the ACP group and was 9 (range, 6-12) in the PBS group (not significant) in experiment 1. 91% of rats in the ACP group developed abscesses, versus 90% in the control group. There were no significant differences in abscess size or number of abscesses. In experiment 2, total adhesion scores in the 2 mL ACP group, 4 mL ACP group, and PBS group were 4 (range, 2-20), 6 (range, 1-11), and 6 (range, 1-18), respectively, (not significant) after 1 week and 3.5 (range, 1-8), 5 (range, 2-15), and 4 (range, 0-9), respectively, (not significant) after 3 weeks. All rats in the 2 mL ACP group and the PBS group and 83% of the 4 mL ACP group had developed abscesses after 1 week. After 3 weeks these percentages were 80, 75, and 73, respectively. There were no significant differences in size or number of abscesses between groups both after 1 and 3 weeks. CONCLUSION: ACP does not reduce adhesion and abscess formation in a rat peritonitis model.

Influence of monocyte-like cells on the fibrinolytic activity of peritoneal mesothelial cells and the effect of sodium hyaluronate.

OBJECTIVE: To determine whether the presence of cells of the monocyte-macrophage system affects the fibrinolytic response of peritoneal mesothelial cells to lipopolysaccharide (LPS) in the presence and absence of sodium hyaluronate. DESIGN: Controlled laboratory experiment. SETTING: Cell cultures in an academic laboratory research environment. PATIENT(S): Human peritoneal mesothelial cells were harvested from patients undergoing a laparotomy for noninfectious reasons and were cultured in vitro. Co-cultures were formed by adding U-937 human monocyte-like cells to a monolayer of mesothelial cells. INTERVENTION(S): After 24 hours, cultures were treated with 10 ng/mL of LPS, and sodium hyaluronate was added in a final concentration of 0.2%. Controls received medium without sodium hyaluronate. MAIN OUTCOME MEASURE(S): After 24 hours' incubation, tissue plasminogen activator (tPA), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor-1 (PAI-1) levels were determined in medium and cell lysates by using ELISA techniques. RESULT(S): In medium of co-cultures, tPA and PAI-1 concentrations were statistically significantly increased compared with the case of monocultures, whereas uPA concentration was statistically significantly decreased. In cell lysates of co-cultures, PAI-1 concentration was statistically significantly increased compared with the case of monocultures, whereas tPA and uPA were unaffected. Treatment with sodium hyaluronate statistically significantly decreased PAI-1 and uPA concentrations in medium of monocultures but decreased uPA concentration only in medium of co-cultures, compared with the case of controls. CONCLUSION(S): Cells of the monocyte-macrophage system modulate the fibrinolytic capacity of LPS treated human peritoneal mesothelial cells and interfere in the hyaluronan-associated changes in mesothelial fibrinolytic capacity.