Preservation of vascular endothelial repair in mice with diet-induced obesity.

INTRODUCTION: Preservation of structural integrity of the endothelial monolayer and maintenance of endothelial cell function are of critical importance in preventing arterial thrombosis, restenosis and atherosclerosis. Obesity has been intimately linked with endothelial dysfunction, and reports of reduced abundance and functional impairment of circulating progenitor cells in obesity have led to the suggestion that defective endothelial repair contributes to obesity-related cardiovascular disease. METHODS: C57BL/6 mice were fed a high-fat diet for either 3 or 6 months to induce obesity; metabolic phenotyping was then carried out before femoral artery wire injury was performed. Endothelial regeneration was then quantified. Mononuclear cells and myeloid angiogenic cells were cultured and characterized for pro-angiogenic properties. RESULTS: No impairment of endothelial regeneration following mechanical endothelial injury in diet-induced obese mice when compared with chow-fed controls was observed, despite the induction of an adverse metabolic phenotype characterized by glucose intolerance and insulin resistance. Dietary-obese mice had increased numbers of circulating myeloid angiogenic cells, which retained normal functional properties including intact paracrine angiogenic effects. CONCLUSION: Preserved endothelial regeneration despite metabolic dysregulation in dietary obese mice suggests that compensatory mechanisms mitigate the deleterious influence of insulin resistance on endothelial repair in obesity.

Lower pain and faster treatment with mechanico-chemical endovenous ablation using ClariVein®.

OBJECTIVES: To assess the efficacy of the ClariVein(®) system of mechanico-chemical ablation of superficial vein incompetence. METHOD: ClariVein(®) treatment uses a micropuncture technique and a 4-Fr sheath to allow a catheter to be placed 1.5 cm from the saphenofemoral junction. Unlike laser (endovenous laser treatment (EVLT)) or radiofrequency ablation (RFA), no tumescence is required. The technique depends on a wire rotating at 3500 r/min causing endothelial damage whilst liquid sclerosant (1.5% sodium tetradecyl sulphate) is infused. The wire is pulled back whilst continuously infusing sclerosant along the target vessel's length. Initially, 8 mL of dilute sclerosant was used, but this was subsequently increased to 12 mL. No routine post-op analgesia was prescribed and specifically no non-steroidal anti-inflammatory drugs. Procedure times and pain scores (visual analogue scale) were recorded and compared to EVLT and RFA. All patients were invited for duplex post-procedure. RESULTS: Fifty-one great saphenous veins and six short saphenous veins were treated and followed up with duplex in the 10 months from July 2011. No major complications or deep vein thrombosis were reported. Duplex showed patency of three treated veins with two more veins having only a short length of occlusion, giving a technical success rate of 91%. Comparison with 50 RFA and 40 EVLT showed procedure times were significantly less for ClariVein(®) (23.0 ± 8.3 min) than for either RFA (37.9 ± 8.3 min) or EVLT (44.1 ± 11.4 min). Median pain scores were significantly lower for ClariVein(®) than RFA and EVLT (1 vs. 5 vs. 6, p < 0.01). CONCLUSION: Mechanochemical ablation with the ClariVein(®) system is safe and effective. After some initial failures, the use of 12 mL of dilute sclerosant results in a very high technical success rate >90% which accords with the limited published literature. Procedure times and pain scores are significantly better than for RFA and EVLT. We await the long-term clinical outcomes.

Cycling related common femoral artery disease: an unusual case in an otherwise healthy male.

A cyclist reported gradual onset, bilateral leg claudication without any atherosclerotic risk factors and with a normal vascular examination. Duplex ultrasound and magnetic resonance angiography revealed bilateral focal stenoses due to coralline plaques of the common femoral arteries (CFAs) but normal proximal and distal vasculature. A surgical endarterectomy with vein patch angioplasty resolved all symptoms. Cyclist's iliac syndrome (whereby mechanical and anatomical factors in competitive cyclists and runners lead to arterial kinking and subsequent intimal hyperplasia, endofibrosis and, ultimately, stenoses of the external iliac arteries) is well documented. This is the first report to our knowledge of a similar process isolated to the CFA.

Agreement between computed tomography and ultrasound on abdominal aortic aneurysms and implications on clinical decisions.

OBJECTIVES: The United Kingdom abdominal aortic aneurysm (AAA) screening programme refers aneurysms with ultrasound (US) diameters of ≥5.5 cm to vascular services for consideration of computed tomography (CT) and intervention. We investigated the discrepancy between US and CT, implications on clinical decisions and question at which stage CT be used. DESIGN/METHODS: AAA USs over 5 years were retrospectively analysed. Patients included had aneurysms measuring ≥5 cm on US with subsequent CT within 2 months (n = 123). Based on maximum US diameters, 44 patients had aneurysms between 5 and 5.4 cm (group I) and 79 patients ≥5.5 cm (group II). Results were cross-referenced. Correlation and limits of agreement were calculated. Two radiologists re-measured 44 pairs of CT/US scans and the inter-observer bias in determining discrepancies between imaging modalities calculated. RESULTS: Mean difference between imaging modalities was 0.21 cm (±0.39 cm, p < 0.001). Limits of agreement were -0.55 to 0.96 cm, exceeding clinical acceptability. Mean difference was higher and significant in group I (0.39 cm, p < 0.001) compared to group II (0.10 cm, p > 0.05). Seventy-percent of group I patients had CT scans revealing diameters of ≥5.5 cm. Inter-observer bias was not significant. CONCLUSION: Significant differences between imaging modalities, more in US diameters of below 5.5 cm, exist. We recommend AAAs measuring ≥5 cm on US should undergo earlier referral to a vascular service and CT.

Tissue engineering of a hybrid bypass graft for coronary and lower limb bypass surgery.

Tissue-engineered blood vessels have largely relied on inelastic scaffolds or biological solutions with uncertain long-term in vivo durability. In this report we present for the first time a hybrid tissue-engineered bypass graft consisting of an elastic scaffold of compliant poly(carbonate-urea)urethane (CPU), incorporated with human smooth muscle cells (SMCs) and endothelial cells (ECs) from the same human source. Human vascular SMCs and ECs were extracted from umbilical cord vessels. The effect of shear stress preconditioning on cell retention on the hybrid bypass graft was investigated under pulsatile arterial flow conditions. Retention of ECs seeded onto CPU precoated with SMCs was significantly improved by a period of shear stress preconditioning, especially when the stress incrementally increased. This is probably because the mechanical stimuli orient cells and increase the release of matrix proteins and attachment factors. The stage is now set for developing a hybrid graft for in vivo studies.

Occult renal impairment is common in patients with peripheral vascular disease and normal serum creatinine.

OBJECTIVE: The incidence of peripheral vascular disease (PVD) and angiography/angioplasty is rising annually. The UK Small Aneurysm Trial and other trials have shown renal function is a predictor of increased mortality and failed infrainguinal bypass despite patent vessels. Renal function is classically assessed by serum creatinine (SCr). However, SCr can be normal despite significant renal impairment. A more sensitive test is creatinine clearance (CrCl) as determined by 24-hour urine collection in combination with SCr. We studied the incidence of renal impairment, as defined by CrCl, in PVD patients with normal SCr. METHODOLOGY: All patients with PVD sufficient to necessitate angiography and normal SCr (< or =120 micromol/l - men; < or =97 micromol/l - women) had their CrCl assessed prior to angiography: using both 24-hour urine collection and the Cockcroft-Gault formula. Various blood tests, a detailed history and examination were performed. A control group of arthritic patients, age and sex-matched with similar SCr, also had their CrCl determined. RESULTS: 65 of 76 patients (86%) with normal SCr had a subnormal CrCl (<100 ml/min) and 49 (65%) had a CrCl below 60 ml/min. In the control group of arthritic patients, the proportion having impaired CrCl was significantly less - 67% below 100 mls/min (p=0.0471) and only 15% below 60 mls/min (p<0.0001). The median and interquartile range CrCl of 52 [38-81] mls/min for PVD patients was significantly worse than for control patients (80 [68-119] mls/min -p<0.0001). The Cockcroft-Gault formula for calculating CrCl did not correlate well with the urinary CrCl for the control group but did for PVD patients (p<0.0001). Factors associated with a significantly reduced CrCl were age of at least 75 years, SCr of at least 85 micromol/l and a history of coronary heart disease (all p<0.05). This had a sensitivity of 88% and specificity of 82% for identifying subnormal CrCl. Statin use was associated with a significantly improved CrCl (p=0.040). CONCLUSION: Most PVD patients with normal serum creatinine have occult, significantly impaired renal function as defined by creatinine clearance. Vascular surgeons should include creatinine clearance in pre-operative assessment of renal function especially in patients over 75 years old, with a history of coronary heart disease or a serum creatinine over 85 micromol/l. The method of determining creatinine clearance could be the Cockcroft-Gault calculation or ideally 24-hour urinary creatinine clearance measurement. This would allow appropriate early referral to a nephrologist for further investigation and management. It is worth noting that statin use seems to be associated with a protective effect on renal function.

Venous thromboprophylaxis in UK medical inpatients.

We prospectively assessed the implementation of venous thromboembolism (VTE) prophylaxis guidelines and the impact of grand round presentation of the data in changing clinical practice. Two NHS teaching hospitals were studied for 24 months from January 2003. Patients were risk stratified according to the THRIFT (thromboembolic risk factor) consensus group guidelines and compared with the recommendations of the THRIFT and ACCP (American College of Chest Physicians) consensus groups. Six months following presentation of the initial results, a further analysis was made to assess changes in clinical practice. 1128 patients were assessed of whom 1062 satisfied the inclusion criteria for thromboprophylaxis. 89% of all patients were stratified as having high or moderate risk of developing VTE. Of these only 28% were prescribed some form of thromboprophylaxis-4% received the THRIFT-recommended and 22% received the ACCP-recommended thromboprophylaxis. The vast majority (72%) received no thromboprophylaxis at all. Reassessment, following data presentation at grand rounds, showed a significant increase to 31% inpatients receiving THRIFT (P<0.0001) and ACCP (P=0.002) recommended thromboprophylaxis. However,the proportion of patients receiving no form of prophylaxis barely changed (72% to 69%: P=0.59). We found a gross underutilization of thromboprophylaxis in hospitalized medical patients. A simple grand-round presentation of the data and recommended guidelines to clinicians significantly increased the proportion of patients receiving recommended thromboprophylaxis but did not increase the overall proportion of patients receiving it. We therefore conclude that a single presentation of guidelines is not enough to achieve the desired levels. Such presentations may only serve to make DVT (deep venous thromboembolism) aware clinicians prescribe prophylaxis more accurately.

Engineering of bypass conduits to improve patency.

For patients with severe coronary artery and distal peripheral vascular disease not amenable to angioplasty and lacking sufficient autologous vessels there is a pressing need for improvements to current surgical bypass options. It has been decades since any real progress in bypass material has reached mainstream surgical practice. This review looks at possible remedies to this situation. Options considered are methods to reduce prosthetic graft thrombogenicity, including endothelial cell seeding and developments of new prosthetic materials. The promise of tissue-engineered blood vessels is examined with a specific look at how peptides can improve cell adhesion to scaffolds.

Cellular engineering of conduits for coronary and lower limb bypass surgery: role of cell attachment peptides and pre-conditioning in optimising smooth muscle cells (SMC) adherence to compliant poly(carbonate-urea)urethane (MyoLink) scaffolds.

OBJECTIVE: We are developing a hybrid arterial bypass graft of compliant poly(carbonate-urea)urethane (MyoLink), endothelial and smooth muscle cells (SMCs). To enhance adhesion of SMCs we assessed various attachment factors and the effect of pre-conditioning on cell retention. METHODS: MyoLink segments were coated with either RGD, superfibronectin, fibronectin, fibronectin-like engineered polymer protein (FEPP), FEPP plus or type 1 collagen overnight. (111)Indium-radiolabelled SMCs were placed onto MyoLink segments for 48 h before being aspirated, then lavaged off. All grafts, aspirates and lavages were counted in a gamma counter. SMC viability on the MyoLink segments was also assessed for viability using the Alamar blue redox assay. Separately, MyoLink grafts lined with radiolabelled SMCs were divided into a pre-conditioned group, exposed to subarterial pulsatile flow whilst another group were held in static culture. After 1-week, grafts were exposed to arterial pulsatile flow whilst radioactivity was assessed using a gamma camera. RESULTS: Only FEPP plus significantly enhanced SMC attachment: mean of 32+/-6% cell attachment compared to 21+/-5% for uncoated control. Cell viability was enhanced by all attachment factors except fibronectin. Pre-conditioning was shown to significantly enhance the retention of SMCs onto the MyoLink once exposed to pulsatile arterial flow: the final attachment was 57+/-7% for the static and 76+/-7% for the pre-conditioned group. CONCLUSIONS: FEPP plus enhances SMC attachment to MyoLink. We believe this is because of its repeating sequences of RGD and its positive charge. Pre-conditioning enhances the retention of SMCs to MyoLink once exposed to pulsatile arterial flow.

Identification of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, arcobacter butzleri, and A. butzleri-like species based on the glyA gene.

Currently, the detection and identification of Campylobacter and Arcobacter species remains arduous, largely due to cross-species phenotypic similarities and a relatively narrow spectrum of biochemical reactivity. We have developed a PCR-hybridization strategy, wherein degenerate primers are used to amplify glyA fragments from samples, which are then subjected to species-specific oligodeoxyribonucleotide probe hybridizations, to identify and distinguish between Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, Arcobacter butzleri, and an A. butzleri-like species. Evaluation of this strategy with genomic DNA from different type strains suggests that this approach is both specific and sensitive and thus may be applicable in a diagnostic assay to identify and differentiate these highly related species.

Campylobacter hyoilei Alderton et al. 1995 and Campylobacter coli Véron and Chatelain 1973 are subjective synonyms.

The taxonomic affiliation of Campylobacter hyoilei was reevaluated by examining a variety of phenotypic and genotypic criteria. Whole-cell protein electrophoresis and a comparison of 66 phenotypic characters revealed that reference strains of C. hyoilei were indistinguishable from Campylobacter coli strains. These data were confirmed by a DNA-DNA hybridization level of 67% between the type strains of the two species. Several species-specific assays based on PCR amplification or probe hybridization further substantiated that C. coli strains and C. hyoilei strains are indistinguishable. It is therefore concluded that C. hyoilei and C. coli represent the same species and that the former name should be regarded as a junior synonym of the latter name.

Characterization of Campylobacter upsaliensis fur and its localization in a highly conserved region of the Campylobacter genome.

Despite increasing recognition of the importance of Campylobacter upsaliensis in human disease little is known about either the virulence properties or genetics of this enteric pathogen. The complete coding sequence of a C. upsaliensis gene has yet to be published. We have cloned and sequenced the complete iron-uptake regulatory (fur) gene from the type strain of this species. The C. upsaliensis fur homolog was isolated from a genomic library of C. upsaliensis ATCC 43954 constructed in phage lambdaGEM-11. The open reading frame identified encodes a polypeptide consisting of 156 amino acids. The 5'-flanking region of the C. upsaliensis fur gene contains 3 putative Fur-binding sequences and two catabolite activator-binding sequences indicating the potential for autogenous and cAMP-mediated regulation, respectively. Primer extension analysis identified a single transcription start site 262 nt upstream from the AUG initiation codon. Sequence analysis indicates that the Fur protein of C. upsaliensis is highly homologous (87% amino acid identity) to Campylobacter jejuni Fur. Furthermore, the arrangement of the lysS and glyA genes downstream of fur is precisely conserved in both C. upsaliensis ATCC 43954 and C. jejuni TGH9011. Using the polymerase chain reaction close linkage of fur with lysS and glyA was also observed among multiple isolates of C. upsaliensis, C. jejuni and C. coli suggesting a possible functional relevance for this conserved genetic arrangement in campylobacteria.