Switching from Intravenous to Oral Tacrolimus Reduces its Blood Concentration in Paediatric Cancer Patients.

BACKGROUND/AIM: Tacrolimus is an essential immunosuppressant for successful allogeneic haematopoietic stem cell transplantation (Allo-HSCT). This study aimed to examine the change in the blood concentration of tacrolimus during switching from intravenous to oral administration in allo-HSCT for paediatric cancer to predict the optimal dosage. PATIENTS AND METHODS: We retrospectively examined the medical records of 63 patients who received allo-HSCT and were administered tacrolimus. To compare bioavailability among different dose ranges, the blood concentration was divided by the dose (C/D). RESULTS: Thirty-nine patients (age range=children 1-15 years, adults 17-67 years) were switched to oral administration of tacrolimus. The C/D after switching was significantly lower in children than in adults (p=0.039). There was a strong positive correlation between age and C/D in children, whereas no correlation was observed in adults. CONCLUSION: In paediatric cancer patients, switching tacrolimus administration route may result in reduced blood concentrations. This tendency is more prominent in younger children.

Effects of Different Chromium Compounds on Hematology and Inflammatory Cytokines in Rats Fed High-Fat Diet.

The aim of the study was to determine how a high-fat diet supplemented with various forms of chromium affects hematological and immune parameters of the blood of rats. The rats received a standard diet or a high-fat diet supplemented with chromium at 0.3 mg/kg body weight (BW) in the form of chromium(III) picolinate, chromium(III)-methionine or nano-sized chromium. Selected hematological parameters were determined in the blood of the rats, including total white blood cell (WBC) count, leukogram, red blood cell (RBC) count, hemoglobin level (HGB), hematocrit (HCT), platelet count (PLT) and platelet percentage (PCT), as well as immune parameters: levels of immunoglobulins A and E (IgA and IgE), interleukin-6 (IL-6), interleukin-2 (IL-2), and tumor necrosis factor α (TNF-α); activity of ceruloplasmin (Cp); and levels of caspase 3 and 8 (Casp3 and Casp8). Feeding rats a high-fat diet increased blood markers of induction of inflammation, ie pro-inflammatory cytokines IL-6 and TNF-α, and also significantly increased IgE. The diet had no effect on the blood count, except for an increase in the number of neutrophils. The chromium compounds tested, particularly Cr-Met and Cr-NPs, stimulated the immune system of the rats, as indicated by increased concentrations of IgA, IgE, IL-2, IL-6, TNF-α, and Cp. Given the increase in inflammatory mediators induced by chromium, it should not be used to mitigate the effects of a high-fat diet. Moreover, chromium picolinate and chromium nanoparticles were shown to increase the content of caspase 3 and 8 in the blood of rats, which indicates a pro-apoptotic effect. The effects of the use of chromium nanoparticles include reductions in the WBC count and in the thrombocyte count (leuko- and thrombopenia). Taking account these data the use of chromium as dietary supplement should be reconsidered.

The incidence of acute oxaliplatin-induced neuropathy and its impact on treatment in the first cycle: a systematic review.

BACKGROUND: Although acute oxaliplatin-induced neuropathy (OXIPN) is frequently regarded to be transient, recent studies have reported prolongation of infusion times, dose reduction and treatment cessation following the first dose of oxaliplatin in quarter of patients. Acute OXIPN is also a well-established risk factor for chronic neuropathy. However, there is underreporting of these parameters during the acute phase (≤ 14 days). This paper systematically reviews the incidence of acute OXIPN and its impact on treatment in the first cycle. METHODS: A systematic literature search was performed using PubMed and Medline. Published original articles were included if they described details about prevalence of oxaliplatin-induced acute neuropathy. RESULTS: Fourteen studies, comprised of 6211 patients were evaluated. The majority of patients were treated with oxaliplatin in combination with leucovorin and fluorouracil (FOLFOX). Most studies used the National Cancer Institute Common Toxicity Criteria to assess acute neuropathy. Acute neuropathy (Grades 1-4) was the most common event with prevalence ranging from 4-98%, followed by haematological (1.4-81%) and gastrointestinal (1.2-67%) toxicities, respectively. Drug regimens, starting dose of oxaliplatin and neuropathy assessment tools varied across studies. In addition, moderate to severe toxicities were common in patients that received a large dose of oxaliplatin (> 85 mg/m2) and/ or combined drugs. The majority of studies did not report the factors affecting acute neuropathy namely the range (minimal) doses required to evoke acute neuropathy, patient and clinical risk factors. In addition, there was no systematic reporting of the number of patients subjected to prolonged infusion, dose reduction, treatment delay and treatment cessation during the acute phase. CONCLUSION: Despite the heterogeneity of studies regarding oxaliplatin starting dose, drug regimen, neuropathy assessment tools and study design, a large number of patients developed acute neuropathy. To develop a better preventive and therapeutic guideline for acute/chronic neuropathy, a prospective study should be conducted in a large cohort of patients in relation to drug regimen, starting/ranges (minimal) of doses producing acute neuropathy, treatment compliance, patient and clinical risk factors using a standardised neuropathy assessment tool.

Monitoring the toxic effects of Pb, Cd and Cu on hematological parameters of Wistar rats and potential protective role of lipoic acid and glutathione.

Heavy metal pollution is a serious environmental and health problem. The negative effects of heavy metals that can enter human body can be reduced by the addition of some supplements. In this study, the effects of lead (Pb), cadmium (Cd) and copper (Cu) on the hematological parameters in Wistar rats in the absence and presence of lipoic acid and glutathione were analyzed. Pb, Cd and Cu intoxication significantly affected the hematological parameters of treated animals. The main effects in the case of Pb and Cd intoxication were decreased values of erythrocytes, hemoglobin and hematocrit (up to 30% and 20% for these two metals, respectively) compared with the control group. Cu intoxication caused decrease in hematocrit, thrombocytes, mean cell volume values (c.a. 15%) and slight decrease in the erythrocyte number, while the value of hemoglobin increased (c.a. 7%). The treatment with lipoic acid and glutathione reduced the toxic effects of these metals in all cases.

Blood gases and energy metabolites in mouse blood before and after cerebral ischemia: the effects of anesthetics.

The levels of blood gases and energy metabolites strongly influence the outcome of animal experiments, for example in experimental stroke research. While mice have become prominent animal models for cerebral ischemia, little information is available on the effects of anesthetic drugs on blood parameters such as blood gases, glucose and lactate in this species. In this work, we collected arterial and venous blood samples from female CD-1 mice before and after cerebral ischemia induced by middle cerebral artery occlusion (MCAO), and we tested the influence of different anesthetic drugs. We found that all of the injectable anesthetics tested (ketamine/xylazine, chloral hydrate, propofol and pentobarbital) caused a decrease in blood pH and partial pressure of oxygen (pO2) and an increase of partial pressure of carbon dioxide (pCO2), indicating respiratory depression. This was not observed with inhalable anesthetics such as isoflurane, sevoflurane and halothane. Significant and up to two-fold increases of blood glucose concentration were observed under isoflurane, halothane, ketamine/xylazine, chloral hydrate, and propofol anesthesia. Lactate concentration rose significantly by 2-3-fold during inhalation of isoflurane and halothane treatment, but decreased by more than 50% after administration of pentobarbital. Permanent cerebral ischemia induced respiratory acidosis (low pH and pO2, high pCO2) which was most prominent after 24 h. Postsurgical treatment with Ringer-lactate solution (1 mL, intraperitoneal) caused a recovery of blood gases to basal levels after 24 h. Use of isoflurane for surgery caused a minor increase of blood glucose concentrations after one hour, but a strong increase of blood lactate. In contrast, anesthesia with pentobarbital did not affect glucose concentration but strongly reduced blood lactate concentrations one hour after surgery. All values recovered at three hours after MCAO. In conclusion, anesthetic drugs have a strong influence on murine blood parameters, which should be taken into account in experiments in mice.

The effects of lactated Ringer's solution (LRS) or LRS and 6% hetastarch on the colloid osmotic pressure, total protein and osmolality in healthy horses under general anesthesia.

OBJECTIVE: To investigate changes in colloid osmotic pressure (COP), total protein (TP) and osmolality (OSM) during anesthesia in horses given intravenous lactated Ringer's solution (LRS) or LRS and hetastarch (HES). STUDY DESIGN: Prospective, clinical trial. ANIMALS: Fourteen horses presented for surgery. Mean age 8.3 ± 1.9 years; mean weight 452 ± 25 kg. METHODS: Horses were premedicated with xylazine intravenously (IV); anesthesia was induced with ketamine and diazepam IV, and maintained with sevoflurane. Butorphanol was administered IV with pre-medications or immediately after induction. Xylazine was administered IV for recovery if necessary. LRS was administered IV to all horses with a target rate of 5-10 mL kg(-1) hour(-1). Half of the horses also received 6% HES, 2.5 mL kg(-1) over 1 hour in addition to LRS. Horses that received LRS only were considered the LRS group. Horses that received both LRS and HES were considered the LRS/HES group. Blood was drawn pre- and post-anesthesia, immediately following induction, and every 30 minutes throughout anesthesia. COP, TP and OSM were measured. RESULTS: COP and TP significantly decreased at similar rates for both treatment groups from pre-anesthetic values. Pre-anesthetic COP was significantly greater in the LRS group when compared to the LRS/HES group pre-, post- and throughout anesthesia. In the LRS group post-anesthetic OSM was significantly different than the pre-anesthesia value and that for the LRS/HES group. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of IV HES (2.5 mL kg(-1), over 1 hour) in combination with LRS does not attenuate the decrease in COP typically seen during anesthesia with crystalloid administration alone. Based on these results, administration of HES at this rate and total volume would not be expected to prevent fluid shifts into the interstitium through its effects on COP.

Mobilization of progenitor cells into the blood by immobilized granulocytic colony-stimulating factor.

The effects of granulocytic CSF immobilized on polyethylenoxide by nanotechnology on the bone marrow and circulating pools of mesenchymal and hemopoietic precursors were studied. Immobilized granulocytic CSF caused the release of progenitor cells of different classes into the blood. The effect of injected immobilized granulocytic CSF was superior to that of nonconjugated granulocytic CSF. Specific activity of oral immobilized granulocytic CSF after oral administration was demonstrated.

Biological functions and therapeutic use of erythropoiesis-stimulating agents: perplexities and perspectives.

Randomized clinical studies, carried out in patients with haematological malignancies and with solid tumours, have consistently demonstrated that treatment with recombinant human erythropoietin (Epo) increases haemoglobin levels, reduces blood transfusion requirements, and improves the quality of life. In addition, identification of erythropoietin receptor (EpoR) expression on many types of non-erythroid and cancer cells has spurred an interest in the extra-haematological activities of Epo itself and other erythropoiesis-stimulating agents (ESAs). Epo and its derivatives have emerged as major tissue-protective cytokines in ischaemic and degenerative damage of cardiovascular, neurological and renal diseases, while their angiogenetic and immunomodulatory properties indicate that their therapeutic potential may extend well beyond erythropoiesis alone. Both preclinical and clinical data, however, have suggested that they may contribute to tumour progression and prejudice survival when administered to anaemic cancer patients, though the results are equivocal and the assumed mechanisms by which tumour growth could be promoted are not fully understood. While these findings offer new perspectives, they nonetheless demand caution in the employment of ESAs. Further, well-designed experimental and clinical studies are warranted.

Radioprotective effects of ATP in human blood ex vivo.

Damage to healthy tissue is a major limitation of radiotherapy treatment of cancer patients, leading to several side effects and complications. Radiation-induced release of pro-inflammatory cytokines is thought to be partially responsible for the radiation-associated complications. The aim of the present study was to investigate the protective effects of extracellular ATP on markers of oxidative stress, radiation-induced inflammation and DNA damage in irradiated blood ex vivo. ATP inhibited radiation-induced TNF-alpha release and increased IL-10 release. The inhibitory effect of ATP on TNF- alpha release was completely reversed by adenosine 5'-O-thiomonophosphate, indicating a P2Y(11) mediated effect. Furthermore, ATP attenuated radiation-induced DNA damage immediate, 3 and 6h after irradiation. Our study indicates that ATP administration alleviates radiation-toxicity to blood cells, mainly by inhibiting radiation-induced inflammation and DNA damage.

[Characterization of cocoa fiber and its effect on the antioxidant capacity of serum in rats]. / Caracterización de la fibra de cacao y su efecto sobre la capacidad antioxidante en suero de animales de experimentación.

OBJECTIVES: The aim of this study was to characterize the physico-chemical properties of cocoa fibre (CF), to analyze its polyphenolic content and antioxidant capacity in vitro, and to investigate the effect of the administration of a polyphenolic extract of this cocoa fiber on the antioxidant capacity of the serum in rats. METHODS AND MATERIALS: Dietary fiber (DF) composition and polyphenolic (PP) content of the cocoa fiber were analyzed. The antioxidant capacity of the CF was determined by means of its reduction power (FRAP) and the capacity to scavenge free radicals (ABTS). To evaluate the bioavailability and the antioxidant capacity in vivo of the phenolic compounds of CF, an extract of these compounds was administred in the stomach of the rats with a gastric probe (100 mg PP/kg), taking blood samples at different time intervals. Sera were analyzed by HPLC to determine the presence/absence of PP or PP-metabolites. In orther to evaluate the antioxidant capacity of the serum FRAP and ABTS methods were used. RESULTS: Cocoa fiber was an excellent source of DF, with a high content of total dietary fiber (TDF), over 60% of the dry matter, made up mainly of insoluble dietary fiber (IDF; 83% of TDF). This fiber had just 1.15% of polyphenols, with low antioxidant activity. After intragastric administration of the PP-rich fraction a fast and measurable absorption of the CF polyphenols was observed, being epicatechin the main PP in blood. The absorption of this PP confers a significant, although transitory increase of the serum antioxidant capacity 10-45 minutes post-gavage; after this time, the antioxidant capacity progressively decreased reaching basal levels after 6 h. CONCLUSIONS: Cocoa fiber can be considered as an excellent source of DF, mainly insoluble dietary fiber; therefore, it could be used as an ingredient in fiber-rich functional foods. Besides the benefits derived from its high fiber content, the CF would provide protection against oxidative damage by means of its content in phenolic compounds (epicatechin) wich are absorbed maintaining the antioxidant properties in vivo.

Effects of methylprednisolone and a biocompatible copolymer circuit on blood activation during cardiopulmonary bypass.

BACKGROUND: Cardiopulmonary bypass (CPB) induces derangements in physiology characterized by activation of blood pathways that may contribute to multiorgan dysfunction. This trial addresses the efficacy of a biocompatible surface alone and in combination with steroids in inhibiting these changes. METHODS: In a factorial design, patients undergoing coronary artery bypass grafting were randomized (four groups; n = 17 per group) to CPB utilizing control circuits or a circuit prepared with a surface modifying active copolymer (SMA-CPB), with or without methylprednisolone (MPSS, 1 g intravenous). Leukocyte and complement activation, cytokine release, and bradykinin generation were measured. Clinical outcomes (blood loss, transfusion, arterial pressure response, and postoperative cardiac and pulmonary functions) were also examined. RESULTS: The SMA-CPB was associated with a significant inhibition of elastase release (p = 0.026) and bradykinin generation (p = 0.027) during CPB. Terminal complement complex (TCC) generation was inhibited as an effect of SMA-CPB (p = 0.047). There was an interaction of SMA-CPB and MPSS to decrease both TCC (p = 0.042) and bradykinin generation (p = 0.028). There were strong effects of MPSS in inhibiting release of interleukin 6 (IL-6) (p = 0.007) and IL-8 (p < 0.001) and tissue plasminogen activator over time (p = 0.009) as well as decreasing peak day 1 creatine kinase (CK, p = 0.015) levels. Clinical effects of MPSS included decreased atrial fibrillation (p = 0.02), improved cardiac index over time, increased pulmonary compliance, and increased insulin need. CONCLUSIONS: This trial suggests a potential beneficial effect for combined strategies to minimize inflammation after CPB. The specific effect of MPSS in decreasing postoperative atrial fibrillation and CK warrants further investigation of its role as a potential myocardial protective agent.

[Indirect electrochemical blood oxidation in the prophylaxis of suppurative-inflammatory complications after transurethral resection of the prostate]. / Nepriamoe élektrokhimicheskoe okislenie krovi v profilaktike gnoino-vospalitel'nykh oslozhnenii posle transuretral'noi rezektsii prostaty.

To investigate efficiency of indirect electrochemical blood oxidation in prophylaxis of infectious complications, 28 patients were treated with 0.06% hypochlorite solution (2 infusions, 2-3 days before TURP). These patients belonged to a high-risk group of infectious complications (80% of them had chronic prostatitis, 8 patients had bladder stones, 8 had undergone cystostomy). The results demonstrated that the complication rate in the above prepared patients fell four times as compared to the control group and the postoperati hospital stay was reduced by 3 days.

Phytoestrogens increase the capacity of serum to stimulate prostacyclin release in human endothelial cells.

BACKGROUND: Both the estrogen receptor (ER) alpha and beta isoforms are expressed in the endothelium. The ER beta has been assigned a crucial role in normal vascular wall function. Prostacyclin has been ascribed a beneficial effect on vessel wall physiology. Isoflavones bind with higher affinity to ER beta. We investigated the hypothesis that their administration to postmenopausal women can promote endothelial prostacyclin production. METHODS: Twenty-five healthy postmenopausal women with mild climacteric symptoms received capsules containing 55 mg/day isoflavones derived from soy and red clover for 6 months. Cultured human umbilical vein endothelial cells (HUVECs) were exposed for 24 h to serum collected before the initiation of therapy and then after 3 and 6 months of continuous therapy. Prostaglandin production was measured in culture medium. RESULTS: In the presence of serum obtained after isoflavone treatment, the prostacyclin production increased significantly from 2.7 +/- 0.5 ng/mg protein at baseline to 3.4 +/- 0.7 ng/mg protein at 3 months (p < or = 0.05), and to 3.8 +/- 0.7 ng/mg protein at 6 months (p < or = 0.05 vs. baseline and 3 months' treatment). CONCLUSIONS: Serum obtained from postmenopausal women treated with isoflavones stimulates the capacity to produce prostacyclin by HUVECs in culture, an effect that could contribute to a beneficial cardiovascular effect of phytoestrogens.

Short-term exposure of cartilage to blood results in chondrocyte apoptosis.

Studies have shown that joint bleeding leads to cartilage degradation independent of concurrent synovitis. We hypothesized that the blood-induced cartilage damage is because of increased chondrocyte apoptosis after short-term exposure of whole blood or isolated mononuclear cells plus red blood cells to cartilage. Human cartilage tissue samples were co-cultured for 4 days with whole blood (50% v/v) or with mononuclear cells plus red blood cells (50% v/v equivalents). Cartilage matrix proteoglycan synthesis ((35)SO(4)(2-) incorporation) was determined after 4 days as well as at day 16 (after a 12-day recovery period in the absence of any additions). To test the involvement of apoptosis a specific caspase-3 inhibitor (acDEVDcho, 0 to 500 micro mol/L) as well as a pan-caspase inhibitor (zVADfmk, 0 to 500 micro mol/L) were added. Chondrocyte apoptosis was evaluated by immunohistochemical staining of single-strand DNA and by terminal dUTP nick-end labeling. Cartilage co-cultured with whole blood as well as mononuclear cells plus red blood cells induced a long-term inhibition of proteoglycan synthesis (74% and 78% inhibition on day 16, respectively). Immunohistochemistry showed a threefold increase in apoptotic chondrocytes in cultures with 50% whole blood as well as with mononuclear cells plus red blood cells. Both the specific caspase-3 inhibitor and the pan-caspase inhibitor partially restored proteoglycan synthesis in the cartilage after blood exposure. This effect was accompanied by a decrease in the number of apoptotic chondrocytes. These data suggest that a single joint hemorrhage (a 4-day exposure of cartilage to 50% v/v blood) results in induction of chondrocyte apoptosis, responsible for the observed inability of the chondrocytes to restore the proteoglycan synthesis during recovery from a short-term exposure to blood. This reduced restoration could eventually lead to cartilage degeneration and ultimately joint destruction.

Inhibition of shear-stress-induced platelet aggregation and phosphotyrosine signaling by GPIIb-IIIa antagonists.

Shear-stress-mediated platelet thrombus formation has been implicated in the pathophysiology of cardiovascular diseases such as acute myocardial infarction and unstable angina. Although previous studies have established that fluid shear forces cause platelet aggregation, a direct comparison of GPIIb-IIIa antagonists used in the treatment of acute coronary syndromes on shear-induced platelet activation has not been reported. Therefore, the objective of the present study was to characterize the effects of the platelet antagonists abciximab, eptifibatide, and tirofiban on shear-mediated platelet activation and aggregation using flow cytometric and Western blotting techniques. Flow cytometric analyses indicated that all three platelet antagonists, when used at concentrations that saturated all GPIIb-IIIa receptors, significantly inhibited platelet aggregate formation and expression of the platelet activation marker p-selectin. None of the antagonists caused increased expression of GPIbalpha or GPIIb-IIIa on the platelet surface compared to untreated controls. Additionally, Western blotting demonstrated that a 72 kDa protein tentatively identified as Syk became phosphorylated in response to shear stress and that its phosphorylation was inhibited by each antagonist. The findings of this study indicate that abciximab, eptifibatide, and tirofiban, though possessing distinct biochemical and pharmacological properties, effectively and equivalently inhibit platelet aggregation, p-selectin expression, and intracellular tyrosine phosphorylation events induced by fluid shear stress.

Quantitative detection of platelet aggregates in whole blood without fixation.

Platelet counting detects lesser degrees of platelet aggregation than conventional aggregometry. In order to prevent progressive platelet aggregation or disaggregation after sampling it is customary to fix blood samples. However fixation may introduce other artefacts. We first compared stability of platelet counts in EDTA-, citrate- and r-hirudin-anticoagulated blood from healthy volunteers. Second, the stability of platelet counts in unfixed EDTA- and hirudin-anticoagulated blood was compared with glutaraldehyde-fixed blood in the same anticoagulants. Third, the effect of in vivo heparin administration on platelet counts in EDTA- and hirudin-anticoagulated blood was studied. Platelet counts within 2 h of collection were significantly higher in EDTA- than in hirudin- or citrate-anticoagulated blood (P = 0.002 vs. hirudin and P = 0.001 vs. citrate). Twenty-four hour counts in hirudin and EDTA were unchanged (P = 0.3 and P = 0.2, respectively, vs. earlier counts). Counts in citrate increased significantly (P = 0.007; n = 10). Platelet counts in fixed blood did not differ significantly from those in unfixed blood. Heparin administered for cardiopulmonary bypass reduced platelet counts in hirudin-anticoagulated blood from (mean +/- 1 standard deviation) 180 +/- 45 to 162 +/- 30 x 10(9) l-1 (P = 0.01; n = 14), without significantly lowering counts with EDTA-anticoagulation, consistent with increased platelet aggregation. Hirudin and EDTA provided stable platelet counts, suggesting that fixation is unnecessary.

[Polychemotherapy-induced changes in the antioxidant state of blood in patients with inoperable cancer of the stomach]. / Izmeneniia antiokislitel'nogo statusa krovi bol'nykh neoperabel'nym rakom zheludka posle provedeniia polikhimioterapii.

Changes in antioxidant characteristics of blood in patients with inoperable tumors of the stomach were followed before and after combination polychemotherapy which comprised intraperitoneal and endolymphatic injection of drugs. General strengthening therapy was given prior to treatment. It was found that enzymatic activity as a component of antioxidant defenses offered by lymphocytes is low in such patients which adversely affects the structural and functional integrity of lymphocyte membranes. After chemotherapy, said function improved significantly showing a tendency towards normalizing. This effect correlated with clinical improvement and better quality of life in patients with inoperable gastric tumors.

Endothelial cell activation by sera containing HLA antibodies is mediated by interleukin-1.

BACKGROUND: It has been suggested that antibodies which are associated with chronic pathological conditions such as chronic rejection and autoimmune diseases have the capacity to activate endothelial cells by induction and up-regulation of adhesion molecules. It has also been suggested that HLA antibodies formed by patients awaiting transplantation can activate endothelial cells. These antibodies include HLA and those that bind to endothelial cells. METHODS: We have further investigated this phenomenon using monoclonal antibodies against HLA class I determinants and sera from aortic valve graft recipients, containing strong HLA antibodies. The effect of 24-hr incubation of antibodies/serum with human umbilical vein endothelial cells (HUVECs) on adhesion molecule expression was measured by flow cytometry. RESULTS: HLA monoclonal antibodies had no effect on ICAM-1 expression on HUVECs. Five of 31 (16%) patients' sera caused strong up-regulation of adhesion molecules (ICAM-1, vascular cell adhesion molecule-1, and E-selectin) but this did not correlate with HLA specificity, IgG, or IgM binding to HUVECs. The activity, found in whole serum and IgG-depleted fractions was inhibited by neutralizing antibodies against interleukin (IL)-1beta and tumor necrosis factor-alpha. Examination of patient sera for presence of IL-1beta demonstrated high levels of IL-1beta in all five sera (range, 30 -500 U/ml) as well as in samples from an additional three patients. CONCLUSION: The ability to activate endothelial cells detected in our patient sera was caused by cytokines and not antibody. Our observation that addition of cytokines to sera before separation into large and low molecular weight fractions demonstrated retention of cytokines in both fractions may be a confounding issue when investigating endothelial cell activation by patients' sera.

An evaluation of the antioxidant activity of a standardized grape seed extract, Leucoselect.

UNLABELLED: Coronary Artery Disease (CAD) remains the major cause of mortality and morbidity in the Western World. The oxidation of low-density lipoproteins (LDLs) by free radicals is associated with initiation of atherosclerosis and therefore, development of CAD. LDLs are protected from oxidation by antioxidants and in times of antioxidant deficiency are more likely to be oxidized. Hypercholesterolaemic patients are at a higher cardiovascular risk and may, therefore, require more antioxidant protection. Increased consumption of red wine containing antioxidants is thought to account for the lower incidence of CAD in Mediterranean countries. Red wine, although rich in antioxidants, is not suitable as routine therapy for prevention of CAD. OBJECTIVE: To evaluate the effects of a capsule formulation of an antioxidant polyphenolic extract of grapes on serum total antioxidant activity and vitamin C and E levels. METHOD: A single-blinded randomised, placebo-controlled cross-over study was undertaken in 20 young volunteers. Subjects were given two capsules containing 300 mg of grape procyanidin extracts (Leucoselect-phytosome) or placebo daily for 5 days. Blood samples were taken at the start of the study and end of the study and assayed for antioxidant activity and vitamins C and E levels. After a washout period of at least 2 weeks, the study was repeated with the second treatment. RESULTS: The extract had no effect on serum vitamins C and E levels but increased serum total antioxidant activity (TAC). On day 5, TAC increased from 408.1+/-22.9 to 453.3+/-453.3 micromol/l trolox equivalents 1 hour postdose. CONCLUSION: The capsules increased serum antioxidant activity but the longer-term clinical implications need to be assessed in further randomised clinical trials.