Key Regulators of Angiogenesis and Inflammation Are Dysregulated in Patients with Varicose Veins.

Varicose veins (VVs) are the most common manifestation of chronic venous disease (CVD) and appear as abnormally enlarged and tortuous superficial veins. VVs result from functional abnormalities in the venous circulation of the lower extremities, such as venous hypertension, venous valve incompetence, and venous reflux. Previous studies indicate that enhanced angiogenesis and inflammation contribute to the progression and onset of VVs; however, dysregulations in signaling pathways associated with these processes in VVs patients are poorly understood. Therefore, in our study, we aimed to identify key regulators of angiogenesis and inflammation that are dysregulated in patients with VVs. Expression levels of 18 genes were analyzed in peripheral blood mononuclear cells (PBMC) using real-time PCR, as well as plasma levels of 6 proteins were investigated using ELISA. Higher levels of CCL5, PDGFA, VEGFC, TGF-alpha, TGF-beta 1, and VEGF-A, as well as lower levels of VEGFB and VEGF-C, were found to be statistically significant in the VV group compared to the control subjects without VVs. None of the analyzed factors was associated with the venous localization of the varicosities. The presented study identified dysregulations in key angiogenesis- and inflammation-related factors in PBMC and plasma from VVs patients, providing new insight into molecular mechanisms that could contribute to the development of VVs and point out promising candidates for circulatory biomarkers of this disease.

Clinical and Pathological Correlations in Chronic Venous Disease.

BACKGROUND: Chronic Venous Disease (CVD) has a high prevalence in the western world. Varicose veins (VVs) are the main signs of this disease that is characterized by important pathological vessel wall changes. The aim of this study is to correlate the main histopathological abnormalities with related clinical issues of CVD. METHODS: A cohort of patients with VVs scheduled for open surgical treatment namely stab avulsion of VVs was recruited. Subsequently, venous tissue from stab avulsion was collected in order to evaluate the following biomarkers: Vascular-Endothelial Growth Factor (VEGF), Protein Gene Product 9.5 (PGP 9.5), Fibronectin (FN), and Matrix Metalloproteinase-9 (MMP-9). The Clinical-Etiology-Anatomy-Pathophysiology (CEAP) criteria were used to classify CVD. RESULTS: Fourteen tissue fragments were processed for histological and immunohistochemical studies. Of these, 43% were from CEAP C2 patients, 36% from CEAP C3 patients, and 21% from CEAP C4 patients. CEAP Class C2 had few to moderate structures positive to VEGF; occasional structures positive to Fibronectin, numerous structures positive to MMP9, few to moderate structures positive to PGP 9.5. CEAP Class C3 had moderate structures positive to VEGF; few to moderate structures positive to Fibronectin; many structures positive to MMP9; few to moderate structures positive to PGP 9.5. CEAP Class C4 had numerous structures positive to VEGF; numerous structures positive to Fibronectin; abundant structures positive to MMP-9; few structures positive to PGP 9.5. CONCLUSIONS: In this study, positive VEGF, FN, and MMP-9 structures were found with increasing trends in relation to the disease staging. VEGF and FN are associated with a progressive increase from C2 to C4. The MMP-9 marker has an important positivity even at early stage of the disease, being higher in CEAP C4 patients. PGP 9.5 decreases in CEAP C4 patients and this is concordant to decreased vein wall innervation.

[ANALYSIS OF SERUM VEGF LEVEL IN PATIENTS WITH VARICOSE VEINS AFTER ENDOVENOUS LASER ABLATION].

The aim of the study was the analysis of serum VEGF level in patients with varicose veins before and after the endovenous laser ablation (EVLA) and to study the role of VEGF in recurrence of the disease. The research involved 216 persons. The study group included 136 patients with varicose veins. Control group included 80 healthy persons. All 136 persons were treated by endovenous laser ablation. To establish the presence of recurrent veins we conducted several observations of patients in period of 1/2-1-3-6-12 months after the treatment and then every 6 month during 3 years. Serum samples were collected from patients' before the operation and 3-6 month after the operation. Serum VEGF level were significantly higher in study group (86,63 pg/ml) comparing to control group 24.10 pg/ml (р 0,01). Mean level of VEGF before the operation were 86.83 pg/ml and in 3 month after the operation decreased to 24.98 pg/ml (р=0,01). In recurrent cases VEGF level were 3 times higher than in non-recurrent cases (р=0,01). There is a direct correlation of VEGF level and recurrence r=0,40, also direct correlation established between the СЕАР clinical class and VEGF level r=0,64.

Erythrocytes properties in varicose veins patients.

Varicose veins (VV) are enlarged veins of the subcutaneous tissue, usually caused by faulty or damaged venous valves leading to impaired blood flow. Blood stasis, excessive clotting disorder and alterations in the vein walls are symptoms of Virchow's triad which may affect the morphotic elements of blood, including erythrocytes. The aim of this study was to investigate alterations in the properties of the erythrocytes taken from varicose veins in comparison to those from antecubital vein of patients with chronic venous disease. The investigation was conducted on whole erythrocytes using spin labeling method in EPR spectroscopy and flow cytometry. The internal viscosity of cells was determined by Tempamine. The conformation state of internal proteins, mainly hemoglobin and membrane proteins was determined by maleimide spin label (MSL, 4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl). The plasma membrane fluidity was measured using two spin labeled fatty acids (5- and 16-doxylstearic acid), while conformational state of membrane protein was measured using two covalently bound spin labels MSL and ISL [4-(2-iodoacetamido)-2,2,6,6-tetramethylpiperidine-1-oxyl]. The osmotic fragility and the shape and size of the erythrocytes were also determined. A decrease in internal viscosity of the erythrocytes from varicose vein was observed. A significant decrease in lipid membrane fluidity indicated by 5-DS, which is located at the polar region of lipid layer was found in the erythrocytes from varicose vein in comparison to normal vein. A significant decrease in the motion of MSL and ISL attached to erythrocyte membrane proteins from varicose vein was found. Changes in the plasma membrane of the erythrocytes from varicose vein were also confirmed by measuring osmotic fragility. These cells were more sensitive to hemolysis than red blood cells from the peripheral blood vein. Meanwhile, no significant differences in size and shape were observed between the erythrocytes taken from varicose veins and those from peripheral veins. In conclusion, the erythrocytes from varicose veins exhibited decreased intracellular viscosity and decreased plasma membrane fluidity. At the same time, conformational changes of membrane proteins and higher osmotic fragility of these cells were found in comparison to the erythrocytes obtained from peripheral veins in the same patients with chronic venous disease. Our findings strongly suggest that presented abnormalities in the erythrocyte plasma membrane may have significant pathophysiological implications, including shortened cell survival and alterations in the hemorheology of the varicose vein blood.

[Concentration of matrix metalloproteinases and magnesium ions in patients with varicose veins of lower limbs]. / Kontsentratsiya matriksnykh metalloproteinaz i ionov magniya pri varikoznoi bolezni ven nizhnikh konechnostei.

The study was aimed at investigating alterations in the concentration of matrix metalloproteinases (MMP-1, MMP-9) and the tissue inhibitor of metalloproteinase-1 (TIMP-1), as well as the level of magnesium ions (Mg2+) as an indicator of connective tissue dysplasia (CTD) in patients presenting with lower limb varicose veins. The study included a total of 110 people. Of these, the Study Group comprised 90 patients with lower limb varicose veins of clinical class C2-C6 (according to the CEAP classification) and the Control Group was composed of 20 apparently healthy volunteers. Samples of peripheral blood were examined. The content of MMP-9, MMP-1 and TIMP-1 in blood serum was determined by means of the quantitative solid-phase immunoenzymatic assay. The concentration of Mg2+ was determined by the colorimetric method. We revealed a statistically significant interrelationship between the concentrations of matrix metalloproteinases and severity of varicose transformation of lower-limb veins, with the highest level of matrix metalloproteinases being observed in patients with cutaneous alterations and trophic ulcers. Determination of the level of matrix metalloproteinases and magnesium ions, characterizing connective tissue dysplasia, makes it possible to predict the development of lower limb chronic venous insufficiency and to evaluate the degree of its severity.

Do blood constituents in varicose veins differ from the systemic blood constituents?

OBJECTIVE: Varicose veins represent one of the most frequent vascular diseases and are in most cases benign. However, advanced disease is frequently associated with complications such as chronic venous insufficiency and superficial vein thrombosis. The pathogenic mechanisms are not well understood. Besides increased venous pressure, it is suggested that local blood constituents trigger various mechanisms responsible for the progression of the disease and its complications. DESIGN: The aim of this study was to investigate the changes in the blood in varicose veins and to compare them with the systemic markers of inflammation and endothelial damage. MATERIALS AND METHODS: Forty patients with primary varicose veins were included in the study. Most patients were class C2. Blood samples were taken from the leg from the tortuous and dilated varicose tributaries of the great saphenous vein and from the cubital vein. RESULTS: The values of basic hematologic tests were comparable between blood samples (varicose vs. systemic). In varicose veins, the following parameters were significantly increased in comparison with systemic blood: hsCRP (3.12 ± 2.18 mg/L vs. 2.04 ± 2.21 mg/L, p = .04), IL-6 (3.54 ± 2.59 pg/mL vs. 2.25 ± 1.27 pg/mL, p = .008), vWF (118.4  ±  27% vs. 83.2  ±  22%, p < .05). D-dimer, in samples taken from the leg varicose veins, was also significantly higher than in the systemic blood (104.3 ± 9.3 ng/mL vs. 89.5 ± 8.3 ng/mL, p = .039). CONCLUSIONS: Some inflammatory markers and indicators of endothelial dysfunction are increased in varicose vein blood. This is most probably the consequence of deteriorated blood flow in dilated and tortuous superficial veins, and increased venous pressure. Damage to the venous wall, which causes a chronic inflammatory response, together with the procoagulant properties of local blood may promote further progression of the disease and thrombotic complications.

[Disabling pathogenetic causes in postthrombotic disease of the lower extremities].

Prospective investigation of etiological and pathogenetic causes of the disabling complications incidence in the lower extremities postthrombotic disease (LEPTHD), influencing activity of these patients, was conducted. The examined patients were divided into two groups, in 62 (58.5%) patients a disability was absent, and in 44 (41.5%) disability was established. Profound clinical examination was conducted, including determination of subfascial pressure on the shin, ultrasound duplex scanning of venous system, electroneuromyography of the lower extremities, estimation of the D-dimer, levels antithrombine-III activity in general and regional blood flow. The leading factors, which causes the LEPTHD patients activity restriction, were determined, basing on the results analysis.

[Algorithm of treatment postthrombotic disease of the lower extremities].

The results of application of pathogenetically substantiated diagnostic algorithm for determination of the treatment method in patients, suffering postthrombotic disease of the lower extremities, are adduced. Using algorithm proposed a clinical state was estimated, subfascial pressure on the shin was determined, ultrasound duplex scanning (USDS) of venous system was conducted, the stimulation electroneuromyography of the shins done, and a level of D-dimer (DD) with activity of antithrombin-III (AT--III) in general and regional blood flow with calculation of its ratio were established. In 33 (31.1%) patients a conservative therapy was conducted, in 34 (32.1%)--a postponed surgical intervention, in 39 (36.8%)--surgical correction of the venous blood flow, in 22 (20.8%)--preparation and closure of trophic ulcers in accordance to the clinic method. Determination of the DD level and the AT-III activity together with data of USDS have permitted to establish differentially the indications for performance of a vein-correcting operative interventions.

[Thrombotic complications of severe forms of varicose disease: modern approach to diagnosis and treatment of hereditary thrombophilia and immunohistochemical peculiarities of vascular wall].

Experience of treatment of 176 patients, suffering thrombotic complications of severe forms of the lower extremities varicose disease (VDLE), was analyzed. In 20 patients, suffering varicothrombophlebitis (VTHPH) in severe forms of VDLE, morphological and immunohistochemical changes in the venous wall and surrounding tissues were studied. There were examined 28 patients, in whom thrombotic complications of the VDLE have had occurred, using diagnostic complex "PLR genetics thrombophilia". Recurrent course of thrombotic complications and coexistence of VTHPH and thrombosis of deep veins have had constitute the main criterion of such patients selection. The groups of patients, suffering severe forms of VDLE, were delineated, depending on thrombotic process localization, differentiated tactics of their surgical treatment was proposed.

Exploring the causal relationship between plasma lipids and varicose veins of lower extremity: A comprehensive two-sample Mendelian randomization study.

Varicose veins of the lower extremities (VVs) is a common chronic vascular disease, with high prevalence rates in some countries; however, their pathogenesis remains unclear. Some studies have identified associations between changes in specific plasma lipid molecules, such as phosphatidylethanolamine (PE), phosphatidylcholine (PC), and sphingomyelin (SM), and the onset of VVs, but due to confounders and reverse causality, the causal relationship remains unclear. Meanwhile, studies on the potential link between other plasma lipids beyond PE, PC, and SM and the risk of VVs in the lower extremities are lacking. This study aimed to explore the potential causal relationship between VVs and plasma lipid levels to provide theoretical insights into the interrelation of plasma lipids and VVs in their occurrence and progression. We conducted a two-sample Mendelian randomization (MR) analysis to assess the potential connection between genetically predicted levels of individual plasma lipids and the risk of developing VVs. We utilized data from a large-scale genome-wide association study involving 7174 Finnish individuals for 179 plasma lipidomes along with VVs genome-wide association study data from 408,455 UK individuals. MR analysis employed methods, such as inverse-variance weighting, weighted median, Bayesian Weighted Mendelian Randomization, and MR-Egger regression. The inverse-variance weighting method was primarily used to assess causality. The validity of the results was demonstrated through sensitivity analysis. In total, 12 lipids were found to have their plasma levels associated with an increased risk of VVs. This includes 3 types of PE, 7 types of PC, and 2 types of phosphatidylinositol. However, no significant causal relationship was found between the plasma levels of 11 types of SM and VVs. These results support the existence of a potential causal relationship between specific types of lipid levels and the risk of VVs, which can provide clues for further studies on biological mechanisms and the exploration of potential therapeutic targets.

Red blood cell deformability and aggregation in chronic venous disease patients with varicose veins.

INTRODUCTION: Red blood cells' (RBC) rheological properties are disturbed in chronic venous disease (CVD). The aim of the study was to compare deformability and aggregation of erythrocytes taken from the varicose vein and the antecubital vein of patients with chronic venous disease. MATERIALS AND METHODS: Blood samples were taken from twelve CVD patients presenting clinical, aetiological, anatomical and pathological elements (CEAP) stages II and III. Blood was sampled from varicose veins and antecubital veins of patients (as control). Deformability and aggregation of RBC were analysed with a Laser-assisted Optical Rotational Cell Analyser (LORCA). RESULTS: A significant increase in deformability was found in varicose vein RBC for shear stress values 4.24, 8.23 and 15.96 Pa as compared to RBC from the antecubital vein. The aggregation index was significantly lower and aggregation halftime was significantly increased for RBC taken from antecubital veins than for RBC from varicose veins. DISCUSSION: In conclusion, RBC taken from varicose and antecubital veins of CVD patients are not entirely rheologically comparable and show different deformability and aggregation. Varicose vein RBC are more deformable and show a higher tendency for aggregation than antecubital vein RBC. Perhaps the deformability of varicose vein RBC has been increased as a compensation mechanism in subjects with CVD, due to increased resistance in their microcirculation.

[Endothelium-dependent factors of pathogenesis of a relapse of varicose disease: a diagnostic concept and treatment policy].

A relapse of lower-limb varicose disease (LLVD), despite the development of surgical achievements in surgical technologies is currently an important problem of present-day phlebology. A considerable factor of its pathogenesis is endothelial dysfunction (ED). The results of clinical studies of "false" relapses (n=32) and the control group were indicative of pronounced ED. Studying the level of leukocytes confirmed a hypothesis on a "leukocytic" trap at the level of the varicose veins of lower extremities. The authors analysed the dynamics of certain markers of ED (CECs, VCAM-1, P-selectin, E-selectin t-RA, endothelin-1), suggestive of the degree of ED severity, revealing a tendency towards normalization of the ED markers on the background of phlebotrophic therapy (Antistax). The carried out study also makes it possible to conclude that the level of endothelemia and biochemical indices of ED may serve as an evaluating marker of the activity of processes of varicose transformation of veins in patients with a relapse of the disease.

Superficial vein thrombosis in patients with varicose veins: role of thrombophilia factors, age and body mass.

OBJECTIVES: To investigate the association of various risk factors including thrombophilia defects, in patients with varicose veins (VVs) and history of episodes of superficial vein thrombosis (SVT). MATERIALS AND METHODS: Two hundred and thirty patients with primary VVs were included in this prospective study. A total of 128 (43 men, age 56 ± 13) had an acute episode or a previous history of SVT, while 102 patients (27 men, age 48 ± 12) did not. Coagulation profile investigation included serum levels of protein C (PC), protein S (PS), anti-thrombin III (AT III), plasminogen (Plg), A(2) antiplasmin (A(2)Apl) and activated protein C resistance (APCR). This was performed at least 3 months after the SVT episode to ensure that the results were not altered. Age and body mass index (BMI) were also assessed. RESULTS: PC deficiency was detected in 3/128 (2.3%), PS deficiency in 19/128 (14.8%), AT III deficiency in 29/128 (22.7%), Plg deficiency in 9/128 (7%), A(2)Apl excess in 3/128 (2.3%) and APCR in 9/128 (7%) patients with SVT and 0/102 (0%), 3/102 (2.9%), 15/102 (14.7%), 6/102 (5.8%), 0/102 (0%) and 1/102 (0.9%) in the control group, respectively. BMI greater than 30 kg m(-2) was associated with SVT. In logistic regression analysis SVT was associated with PS deficiency (odds ratio (OR) 6.7, p = 0.004, 95% confidence interval (CI) 1.83-24.53), obesity (OR 3.5, p = 0.003, 95% CI 1.53-8.05) and age (OR 1.038, p = 0.001, 95% CI 1.01-1.06). CONCLUSIONS: Obesity, age and PS deficiency were found as factors associated with SVT episodes in patients with VVs.

Venous hypoxia: a poorly studied etiological factor of varicose veins.

Venous hypoxia has long been postulated as a potential cause of varicosity formation. This article aimed to review the development of this hypothesis, including evidence supporting and controversies surrounding it. Vein wall oxygenation is achieved by oxygen diffusing from luminal blood and vasa vasorum. The whole media of varicosities is oxygenated by vasa vasorum as compared to only the outer two-thirds of media of normal veins. There was no evidence that differences exist between oxygen content of blood from varicose and non-varicose veins, although the former demonstrated larger fluctuations with postural changes. Studies using cell culture and ex vivo explants demonstrated that hypoxia activated leucocytes and endothelium which released mediators regulating vein wall remodelling similar to those observed in varicosities. Venoactive drugs may improve venous oxygenation, and inhibit hypoxia activation of leucocytes and endothelium. The evidence for hypoxia as a causative factor in varicosities remains inconclusive, mainly due to heterogeneity and poor design of published in vivostudies. However, molecular studies have shown that hypoxia was able to cause inflammatory changes and vein wall remodelling similar to those observed in varicosities. Further studies are needed to improve our understanding of the role of hypoxia and help identify potential therapeutic targets.

[Participation of leucocytes in pathogenesis of primary forms of lower limb chronic venous disease].

AIM: The purpose of the study was to test the hypothesis on participation of WBCs in damaging the venous wall in patients presenting with primary forms of lower limb chronic venous diseases LLCVD . MATERIAL AND METHODS: The study included a total of fifteen consecutively selected patients (13 women and 2 men) diagnosed as having grade C2-C-4 LLCVD according to the CEAP classification. Static loading (30 minutes in the sitting position) was followed by simultaneous sampling of blood from the varicose vein of the cms and ulnar vein. The total blood count including determination of both the absolute values and percentage of blood formed elements was performed using the automated haematological counter «Advia 7¼ («Bayer¼, USA). The obtained findings were statistically processed using the Microsoft Office Excel software by means of the pared two-sample τ-test for the average values. RESULTS: The number of leukocytes and their subpopulations in blood samples obtained from the crural varicose veins turned out to be significantly less as compared with that in blood sampled from the ulnar vein. Thus, blood sampled from the crural varicose veins demonstrated a decrease in the counts of WBC by 9.6% in fourteen (93.3%) patients, that of neutrophils by 4.9% in twelve (80%) patients, that of lymphocytes by 16,8% in fifteen (100%) patients, and that oi monocytes by 24% in twelve (80%) patients. The mentioned differences were statistically significant at a = 0.05. The eosinophilic counts in blood sampled from the upper and lower extremities appeared similar in 66.7% of the examined subjects. In 33.3% of cases the eosinophilic count in blood samples from crural varicose vein was by 16.7% lower than that for blood samples form the ulnar vein. No differences for the rest parameters of the clinical blood count were revealed. CONCLUSION: The absolute lymphocytic count in the blood samples taken after the 30-minute static loading from the crural varicose veins was significantly lower as compared with that in blood sampled form the cubital vein. The counts for RBCs and blood platelets, as well as other qualitative haematological indices (haemoglobin, haematocrit, average volume of the RBC, erythrocytic diameter, etc.) in blood sampled form crural and ulnar veins in the same patient were identical, thus strongly suggesting the lack of either haemodynamic or haemorheological phenomena capable of leading to redistribution of the blood formed elements in varicose veins. Hence a decrease in the counts of leukocytes and their subpopulations in blood sampled from crural varicose veins might be associated with the «leukocytic trap¼ phenomenon.

Elevated sex steroid hormones in great saphenous veins in men.

INTRODUCTION: High serum levels of estradiol are associated with clinical evidence of varicose veins in women; however, the relationship between serum sex steroid hormones and varicose veins in men is unclear. To address this issue, serum levels of testosterone, estradiol, and androstenedione were determined in the great saphenous (GSV) and cubital veins of men with varicose veins. Messenger RNA (mRNA) expression of sex steroid hormones, metabolizing enzymes, and their receptors was investigated in tissue samples of leg veins. METHODS: This prospective study included 40 men, comprising 20 with varicose veins and reflux of the GSV (VM) and 20 with healthy veins (HM). All limbs were assessed by duplex ultrasound scanning of selected superficial and deep leg veins. Blood samples were taken from the cubital vein and from the GSV. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis for sex steroid hormones, their metabolizing enzymes, and receptors in saphenous veins was performed in tissue samples of varicose (n = 6) and healthy veins (n = 6). RESULTS: The VM group had significantly higher (P < .001) mean levels for serum testosterone (44.9 nmol/L; range, 8.8-225.1) and estradiol (242.2 pmol/L; range, 79-941) in varicose saphenous veins compared with cubital veins (testosterone, 15.5 nmol/L; range, 8.4-23.3; estradiol, 93.2 pmol/L; range, 31-147). Moreover, significantly (P < .001) higher mean serum estradiol levels (133.2 pmol/L; range, 63-239) were detected in the saphenous veins of the HM group compared with cubital veins (88.15 pmol/L; range, 37-153). Both groups had similar blood counts and serum androstenedione levels in the upper and lower extremity. Interestingly, qRT-PCR revealed that the mRNA expression of 5alpha-reductase type 1, 5alpha-reductase type 2, 17, 20 lyase, 17beta-hydroxysteroid dehydrogenase (17beta-HSD), aromatase and 3beta-HSD type 2, androgen and estrogen receptor 1 was down-regulated (P < .05) in all samples of varicose veins vs veins obtained from healthy men. CONCLUSION: Elevated serum estradiol and testosterone levels were detected in men with varicose veins and reflux in the GSV compared with the patient's own arm veins. Enzymes and hormonal receptors involved in steroid metabolism were down-regulated in patients with GSV reflux and varicose veins, suggestive of a negative feedback regulation. These data support the notion of a possible causal relationship between sex steroids and varicose veins in men.

Oestradiol levels in varicose vein blood of patients with and without pelvic vein incompetence (PVI): diagnostic implications.

PURPOSE: To assess the difference in the oestradiol levels of blood taken from varicose veins in patients with and without pelvic vein incompetence (PVI). MATERIALS AND METHODS: Women of child-bearing age with symptomatic primary or recurrent varicose veins of the great saphenous vein (GSV) were included in a prospective study. Patients underwent duplex ultrasonography and pelvic vein phlebography. They were divided into a group with PVI (PVI group) and a control group with GSV reflux alone (VV group). Blood samples were collected from the GSV at the sapheno-femoral junction or lower in the thigh as well as from the arm. Oestradiol levels were determined by electroluminescence. RESULTS: Between January and December 2007, 40 women were studied, of which 19 showed phlebographic evidence of PVI (PVI group), while 21 were included in the VV group. Phlebography revealed an incompetent ovarian vein in 14 (74%) patients of the PVI group, dilated uterine and ovarian plexuses in 12 (63%) and an incompetent internal iliac vein in six cases (32%). In the PVI group, the median oestradiol level in GSV samples was 121 pgml(-1) (range: 12-4300), while in the VV group the median level was 75 pgml(-1) (range: 9-1177). In the upper limb, the PVI group patients had a median level of 78 pgml(-1) (range: 15-121) and the VV group patients 68 pgml(-1) (range: 13-568). The ratio of lower limb/upper extremity was significantly higher (p<0.002) in patients of PVI group (median: 1.9; range: 0.7-33) than in those of the VV group (median: 1.1; range: 0.8-13). A threshold ratio of 1.4 showed the highest combined sensitivity and specificity in differentiating patients with PVI from those without. CONCLUSIONS: In patients with varicose veins arising from the GSV, oestradiol levels were significantly higher in the lower limb than in the upper extremity in the subgroup with associated PVI. It may be possible to use this observation as a diagnostic test in patients with suspected PVI. This deserves further study.

Carbonized blood deposited on fibres during 810, 940 and 1,470 nm endovenous laser ablation: thickness and absorption by optical coherence tomography.

Endovenous laser ablation (EVLA) is commonly used to treat saphenous varicosities. Very high temperatures at the laser fibre tip have been reported during EVLA. We hypothesized that the laser irradiation deposits a layer of strongly absorbing carbonized blood of very high temperature on the fibre tip. We sought to prove the existence of these layers and study their properties by optical transmission, optical coherence tomography (OCT) and microscopy. We analysed 23 EVLA fibres, 8 used at 810 nm, 7 at 940 nm and 8 at 1,470 nm. We measured the transmission of these fibres in two wavelength bands (450-950 nm; 950-1,650 nm). We used 1,310 nm OCT to assess the thickness of the layers and the attenuation as a function of depth to determine the absorption coefficient. Microscopy was used to view the tip surface. All fibres showed a slightly increasing transmission with wavelength in the 450-950 nm band, and a virtually wavelength-independent transmission in the 950-1,650 nm band. OCT scans showed a thin layer deposited on all 13 fibres investigated, 6 used at 810 nm, 4 at 940 nm and 3 at 1,470 nm, some with inhomogeneities over the tip area. The average absorption coefficient of the 13 layers was 72 +/- 16 mm(-1). The average layer thickness estimated from the transmission and absorption measurements was 8.0 +/- 2.7 microm. From the OCT data, the average maximal thickness was 26 +/- 6 microm. Microscopy of three fibre tips, one for each EVLA wavelength, showed rough, cracked and sometimes seriously damaged tip surfaces. There was no clear correlation between the properties of the layers and the EVLA parameters such as wavelength, except for a positive correlation between layer thickness and total delivered energy. In conclusion, we found strong evidence that all EVLA procedures in blood filled veins deposit a heavily absorbing hot layer of carbonized blood on the fibre tip, with concomitant tip damage. This major EVLA mechanism is unlikely to have much wavelength dependence at similar delivered energies per centimetre of vein. Optical-thermal interaction between the vein wall and the transmitted laser light depends on wavelength.

Higher prevalence of thrombophilia in patients with varicose veins and venous ulcers than controls.

BACKGROUND: Uncontrolled studies suggest that patients with chronic venous ulceration (CVU) have an increased prevalence of thrombophilia, similar to that observed in patients with deep vein thrombosis. This study compared the nature and prevalence of thrombophilia in patients with varicose veins (VV, CEAP clinical [C] grade C(2) to C(3)) and patients with CVU (C(5) to C(6)) with an age- and sex-matched population without clinical or duplex ultrasound evidence of venous disease. METHODS: Twenty-seven patients with VV, 27 patients with CVU, and 54 age- and sex-matched case controls with no clinical or duplex evidence of lower limb venous disease, underwent testing for factor V Leiden and prothrombin 20210A mutations, antithrombin deficiencies, and levels of antiphospholipid antibodies, homocysteine, protein C and S, and factor VIII, IX, and XI. RESULTS: The overall prevalences of single and multiple thrombophilias were significantly higher in cases than in controls. Specifically, in VV patients, the prevalences of no, single, and multiple thrombophilias were 33%, 52%, and 15%, respectively, compared with 63%, 26%, and 11% in VV controls. In CVU patients, the prevalences of no, single, and multiple thrombophilias was 26%, 30%, and 44%, respectively, compared with 66%, 22%, and 11% in CVU controls. Compared with controls, only factor XI levels were significantly higher in VV patients, and homocysteine and factor VIII, IX, and XI levels were all significantly higher in CVU patients. CONCLUSION: Patients with VV, and particularly CVU, have significantly higher prevalences of single and multiple thrombophilias than age- and sex-matched controls without clinical or duplex evidence of lower limb venous disease. These data support the hypothesis that thrombophilia predisposes to the development of superficial and deep lower limb venous reflux, and so VV and CVU, through the increased occurrence of clinical and subclinical thrombosis.