Single nucleotide polymorphisms in inflammation-related genes are associated with venous thromboembolism.

BACKGROUND: Various genetic risk factors are known to increase the risk of venous thromboembolism (VTE). Increasing evidence suggests a "cross-talk" between the coagulation and inflammatory cascade. Therefore, polymorphisms in genes involved in inflammation may influence susceptibility towards VTE. The aim of the study was to investigate the role of single nucleotide polymorphisms (SNPs) in inflammation genes for susceptibility towards VTE. METHODS: The study group consisted of 108 (47 men and 61 women) Dutch patients with documented VTE and 325 healthy controls from the same geographical area (117 men and 208 women). Odds ratios (OR) and 95% confidence intervals (95% CI) for VTE separately and if indicated by gender were calculated to assess whether genotype and allele frequency were associated with thrombosis. RESULTS: Heterozygosity for SNP -899C/T of the interleukin 1-alpha gene (IL1A -899C/T) was under-represented in VTE patients compared to the control group (OR=0.51, 95% CI 0.32-0.82). The IL6 -174 CC genotype was more frequent in male patients with VTE compared to male controls (OR=4.06, 95% CI 1.43-11.5). Female patients carried significantly more IL13 (intron3) TT genotype (OR=5.60, 95% CI 1.94-18.5) compared to female controls. The allelic frequency of IL4 -589 T allele was significantly increased in female patients (OR=1.72, 95% CI 1.05-2.81) in contrast to men where no differences were observed. CONCLUSION: Four SNPs in inflammatory-related genes of IL1A, IL4, IL6, and IL13 may be associated with VTE. These results need to be confirmed in independent groups with larger number of patients.

Harmonization of fibrinogen assay results: study within the framework of the Dutch project 'Calibration 2000'.

In a Dutch project for harmonization of fibrinogen assays, the commutability of potential calibrators for fibrinogen was assessed by means of a twin-study design, which is, in essence, a multicentre, split-patient sample, between-field-methods protocol. The study consisted of simultaneous analysis of fresh-frozen patient plasmas and three potential calibrators for fibrinogen by 48 Dutch laboratories forming 24 couples. The state-of-the-art intralaboratory standard deviation was used to assess the commutability of the potential calibrators. The potential calibrators were commutable for the Clauss, but not for the prothrombin time (PT)-derived assays. One potential calibrator was used in an attempt to harmonize fibrinogen assay results in a Dutch field study. The interlaboratory coefficient of variation (CV) of three out of four test samples could be reduced significantly using the common calibrator. The average overall CV for the four test samples was 10.3% using the routine measurements and 7.8% using the common calibrator. Despite the reduction in the overall CV, the bias between Clauss and PT-derived assay results in two coumarin test samples could not be eliminated.

Significant reduction in blood loss in patients undergoing minimal extracorporeal circulation.

Several recent studies have shown differences in blood loss and allogeneic transfusion requirements between on-pump and off-pump coronary artery bypass grafting (CABG). Recently a new concept, the mini-extracorporeal circulation, was introduced to minimize the side effects of extracorporeal circulation. Therefore, there are no data comparing the three techniques with special emphasis to blood loss and transfusion requirements. Two hundred and eighty-five patients undergoing first-time coronary artery bypass grafting were retrospectively matched for number of grafts, age and sex. Ninety-five patients underwent surgery with the off-pump CABG (OPCAB) technique, 97 patients using conventional CABG with cold cardioplegia (CCABG) and 93 patients with the mini-extracorporeal circuit with warm blood cardioplegia (MCABG). Blood loss for the CCABG group with a mean loss of 819 +/- 557 mL and the OPCAB group with a mean loss of 870 +/- 768 mL was significant different compared to the MCABG group with a mean loss of 679 +/- 290 mL. The use of units red blood cell units was significantly higher for CCABG group and OPCAB group compared to the MCABG group. On the day of operation the use of platelet concentrate was significantly higher for the CCABG group compared to MCABG group. As a consequence of improvements of several components of the mini heart lung machine, significantly less blood products are needed in MCABG patients. The expected reduced need for transfusion when the pump was completely avoided could not be confirmed in this single retrospective cohort study.

Clinical prospective study of biochemical markers and evoked potentials for identifying adverse neurological outcome after thoracic and thoracoabdominal aortic aneurysm surgery.

BACKGROUND: Neurological deficit after repair of a thoracic or thoracoabdominal aortic aneurysm (TAA/TAAA) remains a devastating complication. The aim of our study was to investigate the clinical value of biochemical markers [S-100B, neurone-specific enolase (NSE) and lactate dehydrogenase (LD)], evoked potentials and their combinations for identifying adverse neurological outcome after TAA/TAAA surgery. METHODS: From 69 patients, cerebrospinal fluid and blood samples for biochemical analysis were drawn after the induction of anaesthesia, during the cross-clamp period, 5 min, 2, 4, 6, 8, and 19 h, respectively, after reperfusion. In addition, continuous perioperative recording of motor-evoked potentials after transcranial electrical stimulation (tcMEP) and somatosensory-evoked potentials was carried out. Furthermore, neurological examinations were performed. RESULTS: In patients with a defined decrease in lower extremity tcMEP during the cross-clamp period, we found that combinations of the serum concentrations of S-100B and tcMEP ratios at 4, 6, and 8 h after reperfusion had a positive and negative predictive value of 100% in predicting adverse neurological outcome after TAA/TAAA surgery. Furthermore, combinations of the serum concentrations of S-100B and NSE or LD at 19 h after reperfusion had both a positive and negative predictive value of 100% in identifying patients with adverse outcome after TAA/TAAA repair. CONCLUSIONS: TcMEP monitoring during TAA/TAAA surgery seems to be an effective but not completely sufficient guide in our protective multi-modality strategy. Combinations of the serum concentrations of S-100B and tcMEP ratios during the early reperfusion period might be associated with adverse neurological complications. Furthermore, biochemical markers could detect central nervous system injury on the first postoperative day and may have prognostic value.

Mini extracorporeal circuit for coronary artery bypass grafting: initial clinical and biochemical results: a comparison with conventional and off-pump coronary artery bypass grafts concerning global oxidative stress and alveolar function.

BACKGROUND: The new concept of mini-extracorporeal circulation (MECC) for coronary artery bypass grafts (MCABG) consists of minimal priming volume, a heparin-coated closed circuit, a centrifugal pump, active drainage, blood cardioplegia and a cell-saving device. The potential organ protective effect of this technique during CABG is unknown. Initial clinical outcomes, oxidative stress, alveolar shunting and need for blood transfusion were investigated for MCABG patients. Sub-sets of these data were compared to outcomes of matched groups of patients operated conventionally (CCABG) and off-pump (OPCAB). METHODS: Data of 184 patients were gathered and analysed from a prospective observational database system. This database consists of the initial experience with the first 114 MCABG operations. Of these, the clinical outcome was investigated. In a subset of 60 MCABGs, need for transfusion was monitored and compared to 60 CCABGs. Serum concentrations of malondialdehyde (MDA), allantoin/urate ratios, shunt fractions and lung epithelium-specific proteins (CC16) were measured as biomarkers of damage during MCABG, CCABG and OPCAB (n =30). RESULTS: Patient groups were similar concerning age, risk and number of distal anastomoses. Clinical outcomes are shown for MCABGs only. During MCABG, need for transfusion was significantly reduced compared to CCABG (p < 0.001). Serum concentrations of MDA and allantoin/urate ratios showed significantly reduced oxidative stress during MCABG compared to CCABG. During MCABG, F-shunts were reduced shortly after surgery. Increased concentrations of pneumoprotein CC16 were measured during CCABG compared to MCABG (data submitted). CONCLUSION: Short-term clinical outcomes of MCABG patients are satisfactory. Compared to CCABG the need for transfusion is significantly reduced when a MECC is used. Oxidative stress parameters show a tendency towards improved global organ protection compared to CCABG. F-shunt fractions and CC16 concentrations suggest reduced alveolar damage during MCABG. In a prospective study, the protective effect of mini-CABG has to be confirmed.

The usefulness of five D-dimer assays in the exclusion of deep venous thrombosis.

D-Dimer measurement is a promising tool in the exclusion of venous thrombosis. New d-dimer assays have been introduced, but need clinical validation. Our objective was to evaluate the clinical usefulness of four relatively new d-dimer assays and a classical ELISA in outpatients suspected for deep venous thrombosis. In 537 patients, participants in a large prospective management study using a clinical probability score and a d-dimer measurement (Tina-quant), additional samples were taken for d-dimer measurement using the Asserachrom ELISA, the VIDAS New, the STA-LIA and the Miniquant assay. Performances of each test were calculated using clinical data during a 3-month follow-up. Thrombosis was detected in 224 patients (42%). The area under the ROC curve was significantly higher for the Tina-quant as compared to the other assays. Using standard cut-off values, sensitivity, negative predictive value (NPV) and specificity of the Asserachrom were 97, 94 and 33%, respectively. For the VIDAS New, values were 100, 96 and 8%, respectively. The Tina-quant showed values of 99, 98 and 41%, respectively, and the STA-LIA 98, 95 and 32%. Values for the Miniquant were 95, 94 and 52%. The d-dimer assays in our study all show a high sensitivity and negative predictive value, but none of the assays reached an NPV of > 98% at standard cut-off values. d-Dimer assays with a low specificity still necessitate additional diagnostic tests in the majority of the patients.

Combination of a normal D-dimer concentration and a non-high pretest clinical probability score is a safe strategy to exclude deep venous thrombosis.

BACKGROUND: Serial ultrasonography is reliable for the diagnosis of deep venous thrombosis in symptomatic patients, but the low prevalence of thrombosis in this group renders the approach costly and inconvenient to patients. We studied the clinical validity of the combination of a pretest clinical probability score and a D-dimer test in the initial evaluation of patients suspected of deep venous thrombosis. METHODS AND RESULTS: Patients with a normal D-dimer concentration (<500 fibrin equivalent units [FEU] microg/L) and a non-high probability score (<3) had no further testing. Patients with a normal D-dimer concentration and a high probability score (> or =3) underwent one ultrasonogram. Serial ultrasonography was performed in patients with an abnormal D-dimer concentration. Patients were followed for 3 months. A total of 812 patients were evaluable for efficacy. Only 1 of 176 patients (0.6%; 95% CI, 0.02% to 3.1%) with a normal D-dimer concentration and a non-high probability score developed thrombosis during follow-up. A normal D-dimer concentration and a high probability score were found in 39 patients; 3 of them (7.7%; 95% CI, 1.6% to 20.9%) had thrombosis at presentation, and one (2.8%; 95% CI, 0.07% to 14. 5%) developed pulmonary embolism during follow-up. In 306 of 597 patients (51.3%) with an abnormal D-dimer concentration, thrombosis was detected by serial ultrasonography. Six patients (2.1%; 95% CI, 0.8% to 4. 4%) developed thrombosis during follow-up. No deaths due to thromboembolism occurred during follow-up. The total need for ultrasonography was reduced by 29%. CONCLUSION: The combination of a non-high pretest clinical probability score and a normal D-dimer concentration is a safe strategy to rule out deep venous thrombosis and to withhold anticoagulation.

S100 protein content of umbilical cord blood in healthy newborns in relation to mode of delivery.

BACKGROUND: Early detection and quantification of brain damage in neonatal asphyxia is important. In adults, S100 protein in blood is associated with damage to the central nervous system. OBJECTIVE: To determine whether S100 protein can be detected in arterial and venous cord blood of healthy newborns and to relate S100 protein concentrations in cord blood to mode of delivery. METHOD: S100 protein levels in umbilical cord blood of 81 healthy infants were determined. RESULTS: S100 protein was present in arterial (median concentration 1.62 micro g/l) and venous (median concentration 1.36 micro g/l) cord blood. Levels were significantly higher in vaginal births (median arterial concentration 1.72 micro g/l; median venous concentration 1.48 micro g/l) than births by caesarean section (1.51 micro g/l and 1.26 micro g/l respectively). CONCLUSION: More research is necessary to determine whether S100 protein is a useful marker in neonatal asphyxia.

Treatment of primary Sjögren's syndrome with D-penicillamine: a pilot study.

BACKGROUND: Up to now no satisfying systemic treatment is available for patients with primary Sjögren's syndrome. METHODS: In a prospective, open study we investigated the effect of D-penicillamine (first three months 250 mg/day, next three months 500 mg/day) on clinical and immunological parameters in 19 patients with primary Sjögren's syndrome and a mean disease duration of 3.8 years. RESULTS: Eight patients had to stop treatment mainly due to severe (reversible) loss of taste. Clinically, a statistically significant increase in basal salivary flow was observed after three months (p<0.05). In addition, improvement was noted in the Schirmer test and stimulated parotid salivary flow after six months, but these differences were not statistically significant. Laboratory values showed a decrease in ESR (p<0.05) and levels of IgA and IgM (both p<0.02) after six months, a decrease in levels of IgA-Rf and IgM-Rf after three months (both p<0.05), and an increase in haemoglobin level (p<0.05). CONCLUSION: From this pilot study we conclude that the treatment of primary Sjögren's syndrome with D-penicillamine has only marginal beneficial effects. Together with its clear side effects this means that D-penicillamine is unsuitable for this indication.

Off-pump versus on-pump coronary artery bypass grafting: oxidative stress and renal function.

OBJECTIVES: Oxidative stress and renal dysfunction occur in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass (on-pump CABG). Whether the same adverse effects also occur during off-pump CABG is the question in this study. METHODS: Forty patients, 27 men and 13 women, undergoing elective CABG were included; 20 patients underwent on-pump CABG and 20 patients underwent off-pump CABG. Renal and ischemia/reperfusion injury parameters were studied, as well as malondialdehyde as a parameter for oxidative stress. RESULTS: The renal function measured as the mean urinary creatinine excretion decreased significantly during surgery for the on-pump CABG group from 7.62+/-4.74 before surgery to 3.07+/-1.49 mmol/l after surgery, whereas no changes occurred in the off-pump CABG group. The mean urinary concentrations of hypoxanthine, xanthine and malondialdehyde expressed as creatinine ratios for the on-pump group increased significantly from 1.92+/-1.36, 6.06+/-3.62 and 0.21+/-0.07 before surgery to 11.88+/-5.77, 13.11+/-6.61 and 0.57+/-0.31 mmol/mol creatinine, respectively at arrival to the intensive care unit (ICU). During the next time-points, the purines and malondialdehyde decreased to 9.21+/-7.46, 7.55+/-3.95 and 0.32+/-0.13 mmol/mol creatinine, respectively after a 20 h stay at the ICU. For the off-pump CABG group, the mean ratios also increased significantly from 1.71+/-1.38, 2.01+/-0.96 and 0.16+/-0.10 before surgery to 4.73+/-3.19, 5.15+/-3.74 and 0.23+/-0.17 mmol/mol creatinine, respectively at arrival to the ICU. During the next time-points, the ratios of xanthine and malondialdehyde decreased to 3.80+/-2.92 and 0.24+/-0.13 mmol/mol creatinine, respectively. The ratio for hypoxanthine reached the highest ratio (6.97+/-5.67 mmol/mol creatinine) after a 9 h stay at the ICU, after which the ratio decreased to 5.98+/-5.56 mmol/mol creatinine after a 20 h stay at the ICU. However, all ratios from the on- and off-pump CABG patients still remained elevated compared with preoperative ratios. In addition, all ratios for the on-pump CABG group were elevated significantly at all time-points for xanthine, at time-points T2 and T4 for hypoxanthine and at time-point T2 for malondialdehyde as compared with the off-pump CABG group. CONCLUSIONS: Only mild signs of oxidative stress and no renal dysfunction were found during and after off-pump CABG compared with on-pump CABG.

Aberrant fibrin formation and cross-linking of fibrinogen Nieuwegein, a variant with a shortened Aalpha-chain, alters endothelial capillary tube formation.

A congenital dysfibrinogenemia, fibrinogen(Nieuwegein), was discovered in a young man without any thromboembolic complications or bleeding. A homozygous insertion of a single nucleotide (C) in codon Aalpha 453 (Pro) introduced a stop codon at position 454, which resulted in the deletion of the carboxyl-terminal segment Aalpha 454-610. The ensuing unpaired cysteine at Aalpha 442 generated fibrinogen-albumin complexes of different molecular weights. The molecular abnormalities of fibrinogen(Nieuwegein) led to a delayed clotting and a fibrin network with a low turbidity. Electron microscopy confirmed that thin fibrin bundles were organized in a fine network. The use of fibrinogen(Nieuwegein)-derived fibrin (fibrin(Nieuwegein)) in an in vitro angiogenesis model resulted in a strong reduction of tube formation. The ingrowth of human microvascular endothelial cells (hMVEC) was independent of alpha(v)beta(3), indicating that the reduced ingrowth is not due to the absence of the RGD-adhesion site at position Aalpha 572-574. Rather, the altered structure of fibrin(Nieuwegein) is the cause, since partial normalization of the fibrin network by lowering the pH during polymerization resulted in an increased tube formation. Whereas factor XIIIa further decreased the ingrowth of hMVEC in fibrin(Nieuwegein), tissue transglutaminase (TG), which is released in areas of vessel injury, did not. This is in line with the absence of the cross-linking site for TG in the alpha-chains of fibrinogen(Nieuwegein). In conclusion, this newly discovered congenital dysfibrinogenemia has a delayed clotting time and leads to the formation of an altered fibrin structure, which could not be cross-linked by TG and which is less supportive for ingrowth of endothelial cells.

The effect of different treatment regimens in reducing fasting and postmethionine-load homocysteine concentrations.

OBJECTIVES: To determine the homocysteine-lowering effect of different treatment regimens on both fasting and postmethionine-load plasma total homocysteine (tHcy) concentrations. DESIGN: Descriptive study of consecutive hyperhomocysteinaemic subjects per treatment regimen. Homocysteine was measured in the fasting state and 6 h after methionine loading, both before and after 8 weeks of vitamin therapy. Hyperhomocysteinaemia was defined as a fasting tHcy and/or increase in tHcy (postmethionine-load minus fasting tHcy concentration) exceeding the 95th percentile of local controls. SETTING: Outpatient clinic of internal medicine of a large non-academic teaching hospital. SUBJECTS: One hundred and seventeen hyperhomocysteinaemic subjects (vascular patients and first-degree relatives). INTERVENTIONS: There were four regimens: pyridoxine, 200 mg; folic acid, 5 mg; combination of folic acid 0.5 mg and pyridoxine 100 mg; and folic acid, 0.5 mg daily. RESULTS: All regimens, except pyridoxine 200 mg, significantly reduced fasting tHcy without differences in the percentage reduction (32-38%). All regimens produced a significant reduction in the increase in tHcy and postmethionine-load tHcy. The reduction in postmethionine-load tHcy was smaller for pyridoxine 200 mg than for combination therapy. No differences were found in the percentage reduction (for both increase in tHcy and postmethionine-load tHcy) between folic acid 5 mg and folic acid 0.5 mg. CONCLUSIONS: Monotherapy folic acid (0.5 mg daily) is the lowest effective therapy for reducing both fasting and postmethionine-load tHcy concentrations, with the same results as high-dose folic acid (5 mg daily). Pyridoxine has no additional value.

Oxidative stress after lung resection therapy: A pilot study.

STUDY OBJECTIVES: To investigate whether oxidative stress occurs following lobectomy and pneumonectomy and to evaluate whether markers of oxidative stress might be of value in the assessment of the diagnosis, course, and prognosis of postoperative complications. DESIGN: A prospective study. SETTING: A specialized thoracic surgical unit in a large referral hospital. PATIENTS: Twenty-eight patients with lung carcinoma undergoing thoracotomy. MEASUREMENTS: Exhaled H(2)O(2) concentrations in breath condensate were measured by spectrophotometry, while malondialdehyde (MDA) levels in urine samples collected every 24 h were measured by reversed-phase, ion-pair high-performance liquid chromatography using ultraviolet detection. RESULTS: Our results show increased H(2)O(2) and MDA levels in lobectomy patients compared with pneumonectomy patients. A strong correlation was found between the levels of H(2)O(2) and MDA. CONCLUSION: The present data support the hypothesis that oxidative stress may occur following pulmonary resection.

The relationship between evoked potentials and measurements of S-100 protein in cerebrospinal fluid during and after thoracoabdominal aortic aneurysm surgery.

OBJECTIVE: This study was performed to correlate the changes in concentration of S-100 protein in the cerebrospinal fluid (CSF) during and after thoracoabdominal aortic aneurysm (TAAA) surgery with the results of somatosensory and motor evoked potential monitoring. METHODS: The study was designed as a prospective study at St Antonius Hospital in Nieuwegein, The Netherlands. The participants were 19 patients who were undergoing elective TAAA surgery. CSF samples for analysis of S-100 protein were drawn after the induction of anesthesia, during the cross-clamp period of the critical aortic segment, after 5 minutes of reperfusion of this segment, during the closure of the skin, and 24 hours after the closure of the skin. In all the patients, continuous intraoperative recording of myogenic motor potentials evoked by transcranial electrical stimulation (tcMEP) and somatosensory potentials evoked by stimulation of the posterior tibial nerve took place to monitor the integrity of the spinal cord. The operative technique consisted of staged or sequential clamping to maximize the beneficial effect of the distal perfusion by the left heart bypass, continuous CSF drainage to keep the CSF pressure below 10 mm Hg, and moderate hypothermia (32 degrees C rectal temperature). We correlated the measured concentrations of S-100 protein in CSF with the results of evoked potential monitoring during surgery and the number of intercostals reimplanted and oversewn. RESULTS: In all the patients, the concentration of S-100 protein was increased in CSF. The highest concentration of S-100 protein was found in the CSF sample taken 5 minutes after reperfusion of the critical aortic segment. There was a good (negative) correlation between the changes in S-100 protein in CSF and the changes in motor evoked potential monitoring during the cross-clamp period. The best (negative) correlation was detected between the S-100 protein elevation in the CSF sample drawn 5 minutes after reperfusion and the tcMEP amplitude reduction during clamping (r = -0.73; P =.007). No relation was found between the S-100 protein dynamics in CSF and somatosensory evoked potential monitoring. A positive (r = 0.58; P =.05) correlation was found between the change in tcMEP amplitude during clamping and the number of reattached intercostals. A moderate to good (r = -0.5 to -0.7; P <.05) correlation between the number of reattached intercostals and the changes in S-100 protein concentration in CSF during TAAA surgery was found. Our data show that transient elevations in S-100 protein after cross clamping are larger in those patients with marked decrease in tcMEP from baseline during the cross-clamp period. CONCLUSION: A correlation is shown between an increasing concentration of S-100 protein in CSF and a reduction in tcMEP amplitude during cross clamping of the aorta. The S-100 protein in CSF seems to be a marker of potential clinical value in the evaluation of the effects of procedures to detect and reduce spinal cord ischemia.

Combination of low-dose folic acid and pyridoxine for treatment of hyperhomocysteinaemia in patients with premature arterial disease and their relatives.

Hyperhomocysteinaemia is an independent risk factor for atherosclerotic disease and venous thrombosis. The optimal homocysteine-lowering vitamin dose and target total homocysteine (tHcy) concentration are currently unknown. We prospectively studied the homocysteine-lowering effect after 8 weeks low-dose combination of folic acid (0.5 mg) and pyridoxine (100 mg) in 49 hyperhomocysteinaemic persons (33 patients with documented premature arterial disease and 16 of their first-degree relatives). Hyperhomocysteinaemia was in both sexes defined as fasting tHcy concentration > 12 micromol/l and/or post-methionine load tHcy concentration > 38 micromol/l. Low-dose vitamin therapy significantly reduced fasting tHcy concentration (median 13.9 to 9.3 micromol/l, reduction 32% (95% CI: 27-37%)) and post-load tHcy concentration (median 55.2 to 36.5 micromol/l, reduction 30% (95% CI: 25-35%)). Fasting tHcy reduction was similar in women and men, as well as in patients and relatives. Post-load tHcy reduction was significantly less in men compared to women (P = 0.04) and in relatives compared to patients (P = 0.03). Although low-dose combination of folic acid and pyridoxine results in a substantial reduction of tHcy concentrations (30-32%) in subjects with hyperhomocysteinaemia, the normalisation percentage to predefined criteria was less impressive (49%).

A protein C pathway (PCP) screening test for the detection of APC resistance and protein C or S deficiencies.

A new automated method for screening defects in the Protein C Pathway (PCP) was evaluated. The "PCP test" is based on a phospholipid-rich Russells viper venom reagent, insensitive to heparin and lupus anticoagulants. To minimize interference from other clotting variables, ratios of the clotting time with and without the addition of a protein C activator were usually determined. Plasma samples from healthy volunteers, patients untreated or on oral anticoagulants, patients with factor V Leiden with and without treatment, and patients with protein C and/or S deficiencies were tested. Mixing patient plasmas 1:1 with individual plasmas deficient in factor V, protein C or S was evaluated for identifying the nature of defects by shortening the screening test. The PCP test was found to be sensitive to APC resistance due to factor V Leiden and by mixing with factor V deficient plasma was also useful despite the effects of oral anticoagulants. Results in the group of patients with previous low protein C or S levels suggest that the method has a better sensitivity to protein C than to protein S deficiency. The automated test was simple to use and gave a between-run coefficient of variation below 3% on normal plasmas.

Homocysteine, methylenetetrahydrofolate reductase polymorphism, antiphospholipid antibodies, and thromboembolic events in systemic lupus erythematosus: a retrospective cohort study.

OBJECTIVE: To examine the relationship between plasma homocysteine, 5,10-methylenetetrahydrofolate reductase (MTHFR) polymorphism, and the risk of arterial and/or venous thrombosis in systemic lupus erythematosus (SLE). METHODS: Plasma homocysteine and MTHFR polymorphism were determined in a retrospective cohort of 175 patients with SLE. Associations between homocysteine levels and antiphospholipid antibodies, renal function, and drug therapy were analyzed. RESULTS: Patients with arterial thrombosis had higher homocysteine concentration than nonthrombotic patients (20.6+/-10.2 vs 13.2+/-6.8 micromol/l; p < 0.001). The crude odds ratio for arterial thrombosis for the highest versus lowest quartile of homocysteine was 4.4 (95% CI 1.3-14.9). After adjustment for other risk factors the odds ratio remained significant (3.7; 95% CI 1.2-13.0). Increased plasma homocysteine was not associated with venous thrombosis. A significant correlation was found between homocysteine and creatinine (r=0.57, p < 0.0001) or the use of methotrexate. Homocysteine levels were higher in patients using corticosteroids and lower in patients using oral contraceptive drugs. This finding may be biased by impaired renal function in patients using corticosteroids and the advice to stop estrogen-containing contraceptives in female patients upon an episode of thrombosis. The distribution of MTHFR genotypes was 50% for homozygous wild type, 42% for heterozygous, and 8% for homozygous mutant, with a similar distribution among patients with or without a history of thrombosis. CONCLUSION: Raised levels of homocysteine are associated with arterial thrombosis in patients with SLE. Mutation in the MTHFR gene does not explain the raised levels, and renal failure is by far the most frequent cause.

Methionine loading test is necessary for detection of hyperhomocysteinemia.

Hyperhomocysteinemia, defined as an elevated concentration of homocysteine in the fasting state or after methionine loading, is an independent risk factor for premature atherosclerosis and venous thrombosis. The role of the methionine loading test (MLT) is, however, controversial. To determine the additional value of the MLT for diagnosis of hyperhomocysteinemia, we prospectively studied 281 patients with premature arterial disease and 148 of their first-degree relatives in the outpatient clinic of a general hospital. Total plasma homocysteine (fasting and 6 hours after methionine loading), folic acid, cobalamin, pyridoxine, and creatinine concentrations were measured. Hyperhomocysteinemia was defined as a fasting homocysteine concentration and/or an increase in homocysteine concentration after methionine loading exceeding the 95th percentile of a healthy control group. Hyperhomocysteinemia was found in 141 (33%) of the 429 subjects: 15% were diagnosed by fasting homocysteine concentration and 18% by MLT. Seventy-eight (55%) of the 141 hyperhomocysteinemic persons were diagnosed only by the MLT. Folic acid was lower in the group with an elevated fasting homocysteine concentration than in those with only an abnormal MLT result (11 versus 15 nmol/L, p = 0.002). Folic acid was significantly negatively correlated, and creatinine significantly positively correlated, with both fasting homocysteine concentration and increase in homocysteine concentration. Negative correlations of cobalamin and pyridoxine with fasting homocysteine concentration were found. In conclusion, the MLT is necessary for diagnosis of hyperhomocysteinemia, because a considerable number of hyperhomocysteinemic persons (55%) remain undiagnosed with the determination of a fasting homocysteine concentration alone.