Rosuvastatin displays anti-atherothrombotic and anti-inflammatory properties in apoE-deficient mice.

Inflammation contributes importantly to all stages of atherosclerosis, including the onset of acute thrombotic complications. In clinical trials, statins are beneficial in the primary and secondary prevention of coronary heart disease. Moreover, statins have been shown to possess several pleiotropic properties independent of cholesterol lowering in experimental settings. Based on these premises, we investigated the anti-inflammatory and anti-atherothrombotic properties of rosuvastatin in vivo, testing its effect on cholesterol and monocyte accumulation, and on adhesion molecules and tissue factor (TF) expression. ApoE-deficient female mice were fed a cholesterol-rich diet containing rosuvastatin (0, 1, 2 or 10 mg kg(-1)d(-1)) for 12 weeks. Treatment with rosuvastatin did not significantly affect either body weight gain or plasma total cholesterol (C) and triglyceride levels. However, rosuvastatin treatment dose-dependently reduced ICAM-1 expression in the aortic valves (V) (up to 40% inhibition, p<0.05) and in the proximal segment of the ascending aorta (AA) (-50%, p<0.001). Similarly, rosuvastatin inhibited VCAM-1 expression in the V (-40%) and in the AA (-35%, p<0.05). Moreover, there was a reduced accumulation of macrophages in the V in a dose-dependent and statistically significant manner (-45%, p<0.01). These anti-inflammatory effects were reflected in a reduction of cholesterol deposition in the entire aorta, both in the free and in the esterified form. Finally, the expression of tissue factor, the most potent pro-thrombogenic agent, was consistently reduced in AA by rosuvastatin treatment (-71%, p<0.001). Altogether, these data demonstrate that rosuvastatin has anti-inflammatory and anti-atherothrombotic activities in apoE-deficient mice that could translate in a beneficial effect on atherogenesis.

An in situ gamma-ray spectrometry intercomparison.

This paper provides the results of an in situ gamma-ray spectrometry intercomparison that was held from 18-21 October 1999, in Grand Junction, CO. This intercomparison was a collaborative effort between the U.S. Department of Energy's Environmental Measurements Laboratory and the U.S. Environmental Protection Agency's Office of Radiation and Indoor Air. It featured measurements of a background location and the Walker Field Calibration Pads. In this paper, the in situ gamma-ray measurements of the background location were compared to soil samples, and the in situ measurements of the Walker Field Calibration Pads were compared to corrected reference values. The results showed that 84% of the in situ gamma-ray measurements of 226Ra, 232Th, and 40K at the background location fell within 20% of the soil sample mean. Similarly, in situ gamma-ray measurements of the Walker Field Calibration Pads showed that 77% of the in situ concentrations fell within 20% of the corrected reference values.

Inhibition of metalloproteinase-9 activity and gene expression by polyphenolic compounds isolated from the bark of Tristaniopsis calobuxus (Myrtaceae).

Excessive breakdown of extracellular matrix by metalloproteinases (MMPs) occurs in many pathological conditions, and thus inhibition of MMP activity might have therapeutic potential. The methanolic extract and the identified compounds from the bark of Tristaniopsis calobuxus Brongniart & Gris (Myrtaceae) were tested on the activity, production, and gene expression of MMP-9. The extract produced a concentration-dependent inhibition (50-95% at 10-50 microg/ml) of MMP-9 activity. The inhibitory activity was retained in the ethyl acetate-soluble fraction (50-95% inhibition at 10-50 microg/ml) which also reduced the release of MMP-9 by mouse peritoneal macrophages up to 80%. In the ethyl acetate-soluble fraction, two active fractions, 5A and 5B were identified. HPLC-MS and NMR analyses of these fractions indicated the presence of gallocatechin, ellagic acid, and its glycoside derivatives. Since the absolute configuration of gallocatechin was not determined, in the next experiments both (+)-gallocatechin (2R,3S) and (-)-gallocatechin (2S,3R) were tested, and (-)-epigallocatechin (2R,3R) was included for comparison. 5A and 5B inhibited MMP-9 secretion, an observation which correlated with the decrease of MMP-9 promoter activity and the downregulation of mRNA levels. All compounds decreased MMP-9 mRNA levels and secretion. Ellagic acid, (+)-gallocatechin and (-)-epigallocatechin, but not (-)gallocatechin inhibited promoter-driven transcription. Thus configuration at C2 (R) of the flavanol seem to be critical for the interaction with the promoter.

Heavy metal contamination in the Taimyr Peninsula, Siberian Arctic.

The Taimyr Peninsula is directly north of the world's largest heavy metal smelting complex (Norilsk, Russia). Despite this proximity, there has been little research to examine the extent of contamination of the Taimyr Peninsula. We analyzed heavy metal concentrations in lichen (Cetraria cucullata), moss (Hylocomium splendens), soils, lake sediment, freshwater fish (Salvelinus alpinus, Lota lota and Coregonus spp.) and collared lemming (Dicrostonyx torquatus) from 13 sites between 30 and 300 km from Norilsk. Element concentrations were low in both C. cucullata and H. splendens, although concentrations of Al, Fe, Cu, Ni and Pb were significantly higher than those in Arctic Alaska, probably due to natural differences in the geochemical environments. Inorganic surface soils had significantly higher concentrations of Cd, Zn, Pb and Mg than inorganic soils at depth, although a lake sediment core from the eastern Taimyr Peninsula indicated no recent enrichment by atmospherically transported elements. Tissue concentrations of heavy metals in fish and lemming were not elevated relative to other Arctic sites. Our results show that the impact of the Norilsk smelting complex is primarily localized rather than regional, and does not extend northward beyond 100 km.

Impairment of CD8+ T suppressor cell function in patients with active systemic lupus erythematosus.

Alteration of T cell suppression function has been recognized in patients with systemic lupus erythematosus (SLE). Recently, CD8(+) T suppressor lymphocytes (CD8(+) Ts) have been generated in vitro by incubating purified CD8(+) T cells with IL-2 and GM-CSF. Using this method, we generated CD8(+) Ts from patients affected by SLE. No major differences were found in the CD8(+) Ts phenotype between SLE patients and healthy subjects. CD8(+) Ts from SLE patients with active disease did not inhibit the anti-CD3 mAb-induced proliferation of autologous PBMC, whereas CD8(+) Ts from SLE patients in remission exerted an inhibitory activity comparable to normal subjects. The inhibitory effect of CD8(+) Ts cells was neither mediated by cytotoxic activity nor by apoptosis induction. Two cytokines, IFN-gamma and IL-6, were found to be responsible for the function of CD8(+) TS: In fact, counteraction of CD8(+) Ts suppression activity was obtained by blocking IFN-gamma with a specific Ab or by inhibiting CD8(+) Ts-mediated IL-6 secretion by an antisense oligonucleotide. Interestingly, CD8(+) Ts from SLE patients showed a peculiar cytokine pattern characterized by an impaired secretion of IL-6 and an increased secretion of IL-12. Thus, it appears that an altered balance between inhibitory (IL-6) and stimulatory (IL-12) cytokines might be responsible for the functional impairment of CD8(+) Ts in SLE patients.

IL-15/IL-15Ralpha intracellular trafficking in human melanoma cells and signal transduction through the IL-15Ralpha.

There are two IL-15 isoforms and eight isoforms for the IL-15Ralpha chain whose biological role is poorly understood. Here, we have analysed the intracellular trafficking of IL-15 and IL-15Ralpha and tried to shed some light on their function(s). In IL-15/GFP CHO transfectants both IL-15 isoforms show nuclear localization. Two melanoma cell lines (MELP and MELREO) spontaneously expressing the IL-15 isoforms, display different intracellular trafficking of the IL-15/IL-15Ralpha complex. In MELP cells only IL-15Ralpha is detected inside the nucleus, whereas IL-15 and IL-15Ralpha assemble at the cell surface and are internalized. Moreover, the transducing molecule TRAF2 co-immunoprecipitates with IL-15Ralpha and may be deflected to TNFRI using anti-IL-15 blocking mAbs and TNF-alpha. By contrast, MELREO cells display IL-15Ralpha and IL-15 nuclear localization but only a partial co-localization of these molecules on the cell surface. In these cells, TRAF2 is strongly associated with IL-15Ralpha and cannot be deflected by any treatment. Since TRAF2 activates the transcription factor NF-kappaB, IL-15 through IL-15Ralpha, could have a role in the control of this pathway. Indeed, anti-IL-15 MaB inhibit the constitutive nuclear localization of NFkappaB and the phosphorylation of its inhibitor Ikappa-Balpha. Thus, IL-15Ralpha controls NF-kappaB activation, however differences in the intracellular trafficking of the IL-15 and/or IL-15Ralpha suggest a different biological role for this complex in MELP versus MELREO cells.

Immune regulatory properties of corticosteroids: prednisone induces apoptosis of human T lymphocytes following the CD3 down-regulation.

Glucocorticoid hormones (GCH) induce apoptosis in PHA-primed peripheral blood T lymphocytes (PBL) and down-regulate membrane-bound proteins involved in the immune response. We have analyzed whether GCH are able to affect the expression of the TCR-associated molecules CD3, CD4, and CD8 on PBL-PHA, and whether the modulation of those receptors is related to the GCH-driven apoptosis of the PBL-PHA. Lymphocytes were cultured with PHA or with PHA plus prednisone (PDN) 10(-3), 10(-6), and 10(-9) M. Then expression of CD2, CD3, CD4, CD8, and CD56 antigens was studied by cytofluorimetric assay using propidium iodide (PI) staining and annexin procedure, and by gel electrophoresis of low molecular weight DNA. PDN, at a pharmacological concentration (10(-6) M), was able to inhibit the CD3 expression on T cells. The kinetics of CD3 decrement and of apoptosis show that the down-regulation of CD3 molecules precedes DNA fragmentation and that the cells lacking CD3 are those prone to PDN-induced apoptosis. The inhibition of CD3 is not related to a transcriptional or posttranscriptional phenomenon, because both PBL-PHA and PBL-PHA-PDN expressed the same amount of intracytoplasmic CD3 molecule. PDN also induced a down-regulation of the CD4 and CD8 molecules that resulted sooner in more intense CD8. In vitro PDN is able to induce apoptosis in PBL-PHA through a down-regulation of CD3 molecules.

Possible differences in the mechanism(s) of action of different glucocorticoid hormone compounds.

Different glucocorticoid hormones (GCH) show differences in the intensity and in the kinetics of their immunomodulating activity. The mechanism(s) of action of GCH is under investigation, but is has been noted that they exert immune activity via the genomic pathway. We have studied the effects of prednisone (PDN), deflazacort (DFC), and dexamethasone (DXM) on the production of cytokines (IL-2, IL-6, TNF-alpha, IL-10) by peripheral T lymphocytes, and the effects on the inhibition of NF-kB DNA binding activity by activated Jurkat cell line. The data obtained show that the three GCH molecules exert an immunosuppression on cytokine production by T lymphocytes and a strong decrease in the nuclear translocation of NF-kB in Jurkat cells; moreover, (a) not all the cytokines investigated were affected, and not with the same intensity, by the three GCH and (b) DXM inhibited the binding activity of NF-kB less than that of DFC and PDN. These data are in agreement with the concept that different GCH compounds might differ in their binding and affinity properties, tissue-specific metabolism, and interaction with transcription factor.

Soy and cholesterol reduction: clinical experience.

A role of vegetable proteins in reducing coronary artery disease risk was postulated as long ago as 1909 in Russia by Ignatowski. The protein hypothesis of atherosclerosis was pursued by many investigators, who studied the possible role of animal vs. vegetable protein in modifying concentrations of plasma lipids and thus cardiovascular disease risk. Over the past 20 y, our research group has examined the potential of a diet based on vegetable protein (in most cases, textured vegetable protein, or TVP) to modify plasma lipid concentrations. Textured products allow administration of a large percentage of protein (up to 50-60% in the product) and are available in a variety of food items. We studied > 1000 patients. An extensive review of the literature indicates that similar findings have been reported by others when administering TVP or TVP-like items to subjects with well-characterized hypercholesterolemia (Fredrickson type II). Data are less consistent for treatment of patients with marginal hypercholesterolemia or hypercholesterolemia already corrected by a standard diet before administration of soy products. The TVP diet, is, however, effective when normolipidemic individuals are made hypercholesterolemic by dietary cholesterol administration. These and other findings suggest that, in man, similar to experimental animals, soy protein may in some way up-regulate LDL receptors depressed by hypercholesterolemia or by dietary cholesterol administration.

Oesophageal transit time and cardiovascular autonomic neuropathy in type 1 (insulin-dependent) diabetes mellitus.

Oesophageal computerized dynamic scintigraphy with 99 mTc was used to evaluate oesophageal motility in type 1 (insulin-dependent) diabetic patients without upper gastrointestinal symptoms. Twenty-nine patients, 10 women and 19 men, mean age 38 +/- 12 yr (range 17-55), mean duration of diabetes 15 +/- 8 yr (range 3-30) and 15 controls were studied. Background or proliferative retinopathy was found in 72.4% of patients, incipient or clinical nephropathy in 48.3% and peripheral neuropathy in 62% of them. In all, oesophagitis and/or other disorders of the upper gastrointestinal tract were excluded by barium studies and endoscopy. Oesophagus scintigraphy with 99 mTc sulphur colloid was performed in each subject after fasting for at least 3 hr in the supine position and repeated after few minutes to assess its reproductivity. The rate of passage of the fluid bolus through oesophagus was analyzed by computer and oesophageal transit time (OTT) for the whole oesophagus was measured by time-activity curves. All diabetic patients were screened for autonomic cardiovascular function by standard tests and, on the base of results, assigned to cardiovascular autonomic neuropathy positive (CVAN-positive) or to cardiovascular autonomic neuropathy negative (CVAN-negative) group. Abnormal oesophageal motility (OTT less than 14 sec as mean +/- 2 SD of controls) was found in 68.7% of CVAN-positive and in 15.4% of CVAN-negative patients (p less than 0.05). CVAN-positive patients resulted older and had significantly longer duration of diabetes than other patients. Furthermore, they showed higher frequency of severe retinopathy, nephropathy, peripheral neuropathy and prolonged OTT compared with CVAN-negative patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Prekallikein and kallikrein inhibitor in liver cirrhosis and hepatitis.

Plasma prekallikrein (kallikreinogen) and kallikrein inhibitor, assayed with the kaolin activable esterase method, have been evaluated in 20 patients with hepatic cirrhosis, in 12 cases with jaundice from acute viral hepatitis, and in 9 normal. A significant reduction of the plasma prekallikrein in cirrhosis has been found. A lowering of plasma prekallikrein has also been observed in viral hepatitis; in this condition, however, the modifications were less important than those obtained in cirrhosis. In three cases of hepatitis, the behaviour of the plasma prekallikrein and kallikrein inhibitor have been controlled during the period of the disease and compared with the behaviour of some conventional parameters, such as serum transaminases and bilirubin. An important increase of the prekallikrein level has been observed during the improvement of hepatitis. These data confirm the implication of the prekallikrein-kallikrein system in severe liver diseases, and indirectly points out the role of the liver in maintaining the physiological balance of the kallikrein system.