Gene and Protein Expression of Chemokine (C-C-Motif) Ligand 19 is Upregulated in Unstable Carotid Atherosclerotic Plaques.

OBJECTIVE/BACKGROUND: The aim was to investigate the expression of genes associated with carotid plaque instability and their protein products at a local and systemic level. METHODS: Carotid plaques from 24 patients undergoing carotid endarterectomy (CEA) were classified as stable or unstable using clinical, histological, ultrasound, and transcranial Doppler criteria, and compared using whole genome microarray chips. Initial results of differentially expressed genes were validated by quantitative reverse transcriptase polymerase chain reaction in an independent group of 96 patients undergoing CEA. The protein product of genes significantly differentially expressed between patients with stable and unstable plaques were analysed by plaque immunohistochemistry and serum protein quantification by enzyme-linked immunosorbent assay on a further independent cohort. RESULTS: Expression of chemokine (c-c-motif) ligand 19 (CCL19) was significantly upregulated in plaques from patients with clinically unstable disease (p < .001). Cathepsin G expression was upregulated in histologically unstable plaques (p = .04). Serum concentration of CCL19 was significantly higher in patients with clinically unstable plaques (p = .02). Immunohistochemical staining for CCL19 demonstrated positive staining in histologically and clinically unstable plaques (p = .03). CCL19 also co-localised with CD3+ T-cell lymphocytes in the core region, around where CCL19 was expressed. CONCLUSIONS: CCL19 is significantly overexpressed in patients with unstable carotid atherosclerotic plaques and may be a possible novel biomarker for identifying high-risk patients in whom more urgent intervention may be indicated.

Microarray-based Gene Expression Profiling of Abdominal Aortic Aneurysm.

OBJECTIVE/BACKGROUND: Microarray-based gene expression profiling studies may detect transcriptional signatures carrying prognostic value in abdominal aortic aneurysms (AAA). A gene expression profiling study was conducted to compare individuals with AAA with screened controls. METHODS: The peripheral blood transcriptome was compared between 12 individuals with AAA and 12 age- and sex-matched controls using microarray. Validation by Taqman real-time quantitative (qPCR) was performed in an independent group as described. Peripheral blood RNA was hybridized to Illumina microarrays, each representing 37,846 genes, allowing comparison of gene expression between cases and controls. Eleven differentially expressed genes were re-quantified by qPCR in the independent group with AAA (n = 95), controls (n = 92), pre- and postendovascular AAA repair (EVAR, n = 31); or open AAA repair (n = 13), AAA wall biopsies (n = 11), and in matched smooth muscle cultures (n = 7). RESULTS: Microarray detected 47 significantly differentially expressed genes in AAA after correction for multiple testing (p < .05). These genes conferred roles in regulation of apoptosis, proteolysis, the electron transport chain, leukocyte migration, and the humoral immune response. Gene quantification in the independent group demonstrated three genes to be downregulated in AAA compared with controls: MSN, PSMB10, and STIM1; however, their expression remained unchanged post-AAA repair. PSMB10 was the only gene conferring a consistent direction of effect in both the discovery and validation analyses (downregulated). EIF3G, SIVA, PUF60, CYC1, FIBP, and CARD8 were downregulated post-EVAR. Expression of all 11 genes of interest was detected in aortic biopsies and matched smooth muscle cultures. CONCLUSION: This study demonstrates differential expression of transcripts in peripheral blood of individuals with AAA, with functional roles in proteolysis, inflammation, and apoptotic processes. These were modulated by aneurysm exclusion from the circulation and expressed in matched aortic biopsies and smooth muscle cultures. These observations further support the key roles for these pathways in the pathogenesis of AAA.

Ultrasonography alone can reliably locate parathyroid tumours and facilitates minimally invasive parathyroidectomy.

INTRODUCTION: Minimally invasive parathyroidectomy (MIP) is performed via a short incision (≤3cm). Previous studies have employed multiple imaging modalities including ultrasonography, sestamibi imaging and/or intraoperative parathyroid hormone assay. We present our eight-year experience of MIP using ultrasonography alone. METHODS: One hundred parathyroidectomies performed by a single surgeon between April 2004 and December 2012 were identified in a prospectively maintained database. All patients underwent ultrasonography including preoperative marking of the lesion by a single radiologist. No other localising diagnostic tests were performed. RESULTS: Of the 100 patients (69% female) who underwent parathyroidectomy, 93 had MIP. The median age of all cases was 58 years (range: 19-90 years). All patients exhibited an elevated parathyroid hormone level (median: 19pmol) in the presence of hypercalcaemia (median: 2.86mmol/l, range: 2.54-3.94mmol/l). Conventional surgery was indicated in seven patients owing to the need for concurrent thyroidectomy. The median operative time was 30 minutes (range: 10-130 minutes). Ultrasonography localised parathyroid tumour position correctly in 98% of patients who underwent MIP, and in 97% across both MIP and non-MIP groups. Postoperative complications requiring treatment included pancreatitis and symptomatic hypocalcaemia. Follow-up review at 6-8 weeks demonstrated that 86% of open cases (6/7) and 94% of MIP cases (87/93) were rendered normocalcaemic. CONCLUSIONS: Our study is the first to demonstrate that the sole use of ultrasonography including preoperative marking can localise parathyroid tumours correctly in 98% of cases suitable for MIP.

Identification of microRNAs associated with abdominal aortic aneurysms and peripheral arterial disease.

BACKGROUND: MicroRNAs are crucial in the regulation of cardiovascular disease and represent potential therapeutic targets to decrease abdominal aortic aneurysm (AAA) expansion. The aim of this study was to identify circulating microRNAs associated with AAA. METHODS: Some 754 microRNAs in whole-blood samples from 15 men with an AAA and ten control subjects were quantified using quantitative reverse transcriptase-PCR. MicroRNAs demonstrating a significant association with AAA were validated in peripheral blood and plasma samples of men in the following groups (40 in each): healthy controls, controls with peripheral arterial disease (PAD), men with a small AAA (30-54 mm), those with a large AAA (over 54 mm), and those following AAA repair. MicroRNA expression was also assessed in aortic tissue. RESULTS: Twenty-nine differentially expressed microRNAs were identified in the discovery study. Validation study revealed that let-7e (fold change (FC) -1·80; P = 0·001), miR-15a (FC -2·24; P < 0·001) and miR-196b (FC -2·26; P < 0·001) were downregulated in peripheral blood from patients with an AAA, and miR-411 was upregulated (FC 5·90; P = 0·001). miR-196b was also downregulated in plasma from the same individuals (FC -3·75; P = 0·029). The same miRNAs were similarly expressed differentially in patients with PAD compared with healthy controls. Validated and predicted microRNA targets identified through miRWalk revealed that these miRNAs were all regulators of AAA-related genes (vascular cell adhesion molecule 1, intercellular cell adhesion molecule 1, DAB2 interacting protein, α1-antitrypsin, C-reactive protein, interleukin 6, osteoprotegerin, methylenetetrahydrofolate reductase, tumour necrosis factor α). CONCLUSION: In this study, circulating levels of let-7e, miR-15a, miR-196b and miR-411 were differentially expressed in men with an AAA compared with healthy controls, but also differentially expressed in men with PAD. Modulation of these miRNAs and their target genes may represent a new therapeutic pathway to affect the progression of AAA and atherosclerosis.

Spontaneous cerebral embolisation in asymptomatic and recently symptomatic patients with TIA/Minor stroke.

OBJECTIVES: Spontaneous embolisation (SE) detected using Transcranial Doppler (TCD) after a Transient Ischaemic Attack (TIA)/Minor stroke is an independent predictor of recurrent stroke. There are, however limited data on the differential prevalence of SE in the first few days/weeks after onset of symptoms. METHOD: 156 consecutive patients (symptomatic n = 123, asymptomatic n = 33) underwent Carotid Endarterectomy (CEA) during an 18 month period and had an accessible window permitting 30 min of pre-operative TCD monitoring. A prospective study was conducted with assessors blinded to clinical status. RESULTS: Spontaneous embolisation was detected in 31 symptomatic patients (25%) of which 1/1 (100%), 14/35 (40%), 8/37 (22%) and in 8/50 (16%) patients presented within 48 h, 3-7 days, 8-14 days and >14 days respectively from the index clinical event. SE occurred in only 6% of asymptomatic patients. Out of 31 symptomatic patients with SE, seven (22.6%) suffered recurrent cerebrovascular events following admission as opposed to 11/92 patients (11.9%) who had no evidence of spontaneous embolisation after admission (OR 2.2 (95% CI 0.8-6.1))(P = 0.2) CONCLUSION: Patients presenting for CEA in the hyperacute period after onset of TIA/Minor stroke have a high incidence of SE. Patients with SE had a 23% risk of recurrent cerebrovascular events. These data support the current drive towards expedited CEA in recently symptomatic patients.

Telomere length dynamics in vascular disease: a review.

INTRODUCTION: Telomeres are specialised DNA-protein complexes which cap the ends of linear chromosomes serving to maintain DNA integrity during cell division. Telomere length naturally shortens with successive cell divisions and represents a cellular marker of biological age. This paper aims to provide an overview of telomere biology and review the evidence for any association between vascular surgical conditions and short telomere length. METHODS: A systematic review of the literature was performed using the search terms 'telomere' and 'vascular'. RESULTS: Considerable associations between a shorter mean telomere length and coronary heart disease have been observed. This finding extends to vascular disease risk factors including age, sex, smoking, obesity, hypertension and diabetes. Vascular diseases including abdominal aortic aneurysm, peripheral vascular disease and carotid disease were also associated with shorter telomere lengths but evidence was limited to a small number of studies. There were no reports of short telomere length associated with varicose veins or arterio-venous malformations suggesting a novel area for further investigation. CONCLUSION: Multiple associations between short telomere length and vascular disease characterised by atherosclerosis suggest a possible link between telomere attrition and disease mechanisms. Further studies are warranted to validate and define the role of telomeres in vascular disease pathogenesis.