Experimental models using rats suggest that decreased endothelial nitric oxide synthase (eNOS) activity in old males promotes renal atherosclerosis, whereas females are protected. We aimed to explore whether aging alters aortic arginine uptake by CAT-1, the selective arginine supplier to eNOS in rats. Arginine uptake by freshly harvested aortic rings from young males (9 weeks) was significantly higher than in young females. Old males (18 months) exhibited a significant decrease in arginine transport compared to young males, whereas no differences were observed between old and young females. Cationic amino acid transporter-1 (CAT-1) abundance remained unchanged in all experimental groups. The abundance of protein kinase C (PKC), a CAT-1 inhibitor, was significantly augmented in old versus young males while no differences were detected between old and young females. Phosphorylated PKC was significantly increased in old rats of both sexes. Tocopherol, a PKC inhibitor, produced a significant increase in arginine transport in old males only. In conclusion, aortic arginine transport by CAT-1 is attenuated in old male rats through upregulation of PKC. In old females, aortic arginine transport is protected from the effects of PKC by an unknown mechanism.

Much effort has been spent to reduce the number of amputees within the last two decades, but it remains unclear how effective the different strategies have been. We analyzed the prevalence of amputations in inpatient cases in the federal statistics. Detailed lists of all amputations coded as major amputations (OPS 5-864) and minor amputations (OPS 5-865) performed in 2005 and 2006, divided into the 4th and 5th number of the OPS-code, were provided by the Federal Statistical Office. In 2006, a total of 62,880 amputations affecting the lower extremities (2005: 63,005) were performed in Germany. Because of multiple amputations within a single case the corresponding cases amounted to 55,705 in 2006 and 55,689 in 2005. Based on these data age-adjusted incidence rates of major amputations per 100,000 inhabitants increased from 2 at the age of < 50 years to 201 at the age of > 80 years. For minor amputations the prevalence rates increased from 4 at the age of < 50 years to 209 at the age of > 80 years. It can be assumed that peripheral arterial disease or neurovascular disease as the underlying disease necessitating the amputation were present in 74.9% of all inpatient cases who finally underwent amputation. There were 12.9% with non-vascular or non-diabetic reasons for amputations. In patients presenting with gangrene, the rate of minor amputations decreased with age, whereas the rate of major amputations increased, especially within the 8th to 10th decades of life. In conclusion, amputations affecting the lower limbs are still a relevant problem in Germany. At the time of an aging German population it has to be an important goal to lower or at least to stabilize the rate of amputations. The DRG statistics enable the Federal Ministry of Health and health politics to monitor amputation rates easily.

Peripheral arterial disease (PAD) is common in patients with severe coronary artery disease (CAD) and is considered a relative contraindication to external enhanced counterpulsation (EECP), but there are no data that define the efficacy and safety of EECP in patients with PAD. The International EECP Patient Registry (IEPR) was used to compare initial posttherapy and 2-year follow-up clinical outcomes and adverse event rates in patients with and without PAD. From January 2002 to October 2004, 2126 patients were enrolled in the IEPR, of whom 493 (23%) had a history of PAD. Immediately following EECP, the reduction in angina (≥ 1 Canadian Cardiovascular Society class) was similar in patients with and without PAD (76.6% vs 79.0%, p = 0.27) as was improvement in the Duke Activity Score Index (DASI) score (+4.7% vs +6.1%, p < 0.001). Both angina reduction and DASI score improvement were sustained at 2 years. PAD patients discontinued EECP more frequently (12.0% vs 8.5%, p < 0.05), but lower extremity ulceration did not occur more frequently in patients with PAD (3.7% vs 2.7%, p = 0.26). Rates of death (17.1% vs 8.6%, p < 0.001) and myocardial infarction (9.5% vs 5.0%, p < 0.001) were, as expected, higher in patients with PAD compared to patients without PAD at 2 years. In conclusion, while PAD patients constitute a high-risk cohort with known higher adverse event rates, EECP led to similar short- and longterm improvements in angina and quality of life for individuals with PAD compared to those without PAD.

Owing to recent progress in analytical techniques, metallomics are evolving from detecting distinct trace metals in a defined state to monitor the dynamic changes in the abundance and location of trace metals in vitro and in vivo. Vascular metallomics is an emerging field that studies the role of trace metals in vasculature. This review will introduce common metallomics techniques including atomic absorption spectrometry, inductively coupled plasma-atomic emission spectrometry, inductively coupled plasma-mass spectrometry and X-ray fluorescence spectrometry with a summary table to compare these techniques. Moreover, we will summarize recent research findings that have applied these techniques to human population studies in cardiovascular diseases, with a particular emphasis on the role of copper in these diseases. In order to address the issue of interdisciplinary studies between metallomics and vascular biology, we will review the progress of efforts to understand the role of copper in neovascularization. This recent advance in the metallomics field may be a powerful tool to elucidate the signaling pathways and specific biological functions of these trace metals. Finally, we summarize the evidence to support the notion that copper is a dynamic signaling molecule. As a future direction, vascular metallomics studies may lead to the identification of targets for diagnosis and therapy in cardiovascular disease.

The objective of this study was to analyze the use of sirolimus-eluting stent (SES) placement for the treatment of renal artery in-stent restenosis (RA-ISR). The optimal treatment of RA-ISR has not been fully elucidated to date. We retrospectively analyzed consecutive patients from our institution who underwent treatment of RA-ISR with a SES from May 2004 to June 2006. Using duplex ultrasound, RA-ISR (> 60% diameter) was determined by peak systolic velocity (PSV) > 300 cm/s and renal aortic ratio (RAR) > 4.0. Renal function (creatinine) and blood pressure were measured at baseline and follow-up. SESs were implanted in 16 patients (22 renal arteries) during the study period. The study cohort was predominantly female (75%) with a mean age of 68 % 12 years. RA-ISR was treated with SESs with a mean diameter of 3.5 mm and mean length of 17.9 ± 3.8 mm. The mean post-dilation balloon diameter was 4.8 ± 0.6. The baseline renal artery PSV was 445 ± 131 cm/s with a mean RAR of 5.0 ± 1.6. Follow-up information was available in 21 renal arteries. During a median follow-up of 12 months (range: 9–15 months), 15 renal arteries (71.4%) developed recurrence of ISR by ultrasonographic criteria. Univariate analysis revealed that female sex was an independent predictor of recurrence of ISR after SES implantation (p < 0.05). In conclusion, placement of a SES for the treatment of ISR in renal arteries is associated with high initial technical success but significant restenosis on duplex ultrasonography at follow-up.

Varicose veins (VVs) are associated with lifestyle-limiting symptoms and complications. Patients who fail compression therapy are candidates for more invasive treatments. This study evaluates the efficacy and safety of endovenous foam sclerotherapy (EFS) for the treatment of VVs in a US academic center. We reviewed medical records of a consecutive cohort of patients who underwent EFS over a 2-year period. The primary outcome measure was obliteration of VVs. The secondary outcome measures were symptomatic improvement, ulcer healing, recurrence, and adverse events. A total of 166 patients (217 legs) underwent EFS for pain (81%), pruritis (41%), swelling (17%), ulcerations (17%), thrombophlebitis (14%), and varix rupture (3%). Complete (65%) or near-complete (34%) obliteration was achieved in 215 (99%) legs after one injection. Additional injections achieved complete obliteration in 39 of 53 legs. Ninety-three percent (27/29) of active ulcers healed or were decreasing in size. Five ulcers and 11 VVs recurred. Common adverse events included pain and hyperpigmentation. Thrombosis, hematoma, skin necrosis, and neurologic events were rare. In conclusion, EFS appears to be a safe and effective outpatient therapy for the treatment of symptomatic and complicated VVs.

We determined the effect of claudication pain on temporal and spatial gait characteristics, and on ambulatory symmetry at preferred and rapid self-selected walking paces in patients with unilateral peripheral arterial disease (PAD). Twentyeight patients with PAD limited by intermittent claudication were studied. Patients ambulated at their preferred and rapid paces over a 7.3-meter portable gait mat system while they were pain-free and after experiencing claudication pain. The order of the pain-free and painful walking trials was randomized, and the following gait parameters were obtained: velocity, cadence, stride length, swing time, stance time, single-support time, and double-support time. During the selfselected rapid pace, patients walked 3% slower (p = 0.020) while in pain due to a 3% shorter stride length (p < 0.001), and they were in double-stance longer (p = 0.024). Claudication pain in the symptomatic leg resulted in an increase in single-stance (p = 0.007). Furthermore, gait became asymmetrical with pain, as the symptomatic leg spent a higher percentage of the gait cycle in the swing phase (p < 0.01) and lower percentages in stance (p < 0.01) and single-stance (p < 0.01) than the asymptomatic leg. Ambulation was symmetrical for all measures during the pain-free trial. In conclusion, claudication pain slows ambulatory velocity at preferred and rapid paces, and increases asymmetry when ambulatory function is challenged with rapid walking. The reduced ambulatory speed with the development of claudication pain may be an adaptation to elicit a safer and less destabilizing gait pattern.

Ultrabiomicroscopy is a novel high-frequency (55 MHz) ultrasound technique that could be used to non-invasively measure the vessel wall and separate the intima–media complex into measurements of intima and media thickness. Since no previous study has measured intima and media thickness separately in vivo in patients with coronary heart disease (CHD), the aim of the current study was to measure intima and intima–media thickness of the radial and the anterior tibial arteries among patients with CHD and healthy subjects (HS). Thirty-two patients with CHD and 46 HS underwent investigations with ultrabiomicroscopy measurements of the radial and anterior tibial arteries. Patients with CHD showed a 19% increase in intima thickness of the radial artery compared with HS (0.088 ± 0.024 mm versus 0.074 ± 0.015 mm; p < 0.015), whereas no difference was seen in media thickness. There were no differences in intima or media thickness within the anterior tibial arteries. In conclusion, CHD is associated with thickening of the intima of the radial artery whereas media thickness was unchanged compared with HS. Assessment of intima thickness by highfrequency ultrasound may provide a tool for non-invasive early detection of atherosclerosis.

Cholinergic angiogenesis is mediated by an endothelial nicotinic acetylcholine receptor (EC nAChR). Short-term administration of nicotine stimulates angiogenesis via EC nAChRs. The long-term effects of nicotine upon cholinergic angiogenesis are unknown. The objective of this study was to determine whether chronic nicotine exposure blunts angiogenesis. We exposed C57/Bl6 male mice (n = 42) to nicotine (200 µg/ml drinking water) or vehicle for 8 or 16 weeks. Subsequently, hindlimb ischemia was induced by ligation of the left femoral artery. After surgery, animals in the vehicle-treated group were re-randomized to vehicle (vehicle group) or nicotine (acute exposure group) for 2 weeks; whereas animals that had been previously treated (for 8 or 16 weeks with nicotine) continued to receive nicotine (8 WK or 16 WK groups). After 2 weeks, animals were sacrificed for immunohistochemical, gene expression, and angiogenesis studies. Capillary density of the ischemic hindlimb was increased by nicotine in naïve animals (vehicle vs acute exposure: 2.40 ± 0.09 vs 2.82 ± 0.10 capillaries/myocyte, p < 0.05). However, prior exposure to nicotine for 16 weeks (16 WK) abolished the effects of nicotine to increase capillary density in the ischemic hindlimb (acute vs 16 WK: 2.82 ± 0.10 vs 2.47 ± 0.03 capillaries/ myocyte; p < 0.05). The impairment of cholinergic angiogenesis was associated with a reduction in nAChR expression and plasma VEGF levels. Chronic exposure to nicotine impaired capillary sprouting of aortic segments ex vivo (vehicle vs 16 WK: 0.303 ± 0.029 vs 0.204 ± 0.017 mm², p < 0.05, n = 3 in each group). In conclusion, the current study shows for the first time that chronic exposure to nicotine impairs cholinergic angiogenesis, an effect mediated by downregulation of the vascular nAChR, and attenuation of nicotine-induced VEGF release. These studies may explain the impairment in angiogenic processes observed in long-term smokers.