Statin use and risk of new-onset diabetes: A meta-analysis of observational studies.

BACKGROUND AND AIMS: Meta-analyses of randomized control trials investigating the association between incident diabetes and statin use showed an increased risk of new-onset diabetes (NOD) from 9% to 13% associated with statins. However, short follow-up period, unpowered sample size, and lack of pre-specified diagnostic criteria for diabetes detection could be responsible of an underestimation of this risk. We conducted a meta-analysis of published observational studies to evaluate the association between statins use and risk of NOD. METHODS AND RESULTS: PubMed, EMBASE and MEDLINE databases were searched from inception to June 30, 2016 for cohort and case-control studies with risk of NOD in users vs nonusers, on ≥1000 subjects followed-up for ≥1 year. Two review authors assessed study eligibility and risk of bias and undertook data extraction independently. Pooled estimates were calculated by a random-effects model and between-study heterogeneity was tested and measured by I2 index. Furthermore, stratified analyses and the evaluation of publication bias were performed. Finally, the meta-analysis included 20 studies, 18 cohort and 2 case-control studies. Overall, NOD risk was higher in statin users than nonusers (RR 1.44; 95% CI 1.31-1.58). High between-study heterogeneity (I2 = 97%) was found. Estimates for all single statins showed a class effect, from rosuvastatin (RR 1.61; 1.30-1.98) to simvastatin (RR 1.38; 1.19-1.61). CONCLUSIONS: The present meta-analysis confirms and reinforces the evidence of a diabetogenic effect by statins utilization. These observations confirm the need of a rigorous monitoring of patients taking statins, in particular pre-diabetic patients or patients presenting with established risk factors for diabetes.

Telomere shortening over 6 years is associated with increased subclinical carotid vascular damage and worse cardiovascular prognosis in the general population.

INTRODUCTION: Leucocyte telomere length (LTL) is an important determinant of telomere function and cellular replicative capacity. The aim of the present study was to examine prospectively the associations between telomere shortening (TS) and both the progression of atherosclerosis and the incidence of cardiovascular events (CVEs). MATERIALS AND METHODS: Leucocyte telomere length was measured by quantitative polymerase chain reaction to determine the ratio of telomere length to single-copy gene (T/S) in 768 subjects (462 female and 306 male) enrolled in a large general population survey [the Progressione della Lesione Intimale Carotidea (PLIC study)]. Common carotid artery intima-media thickness was determined at baseline and after 6 years of follow-up, and the associations between TS and the progression of atherosclerosis and incidence of CVEs were evaluated. RESULTS: Mean LTL was 1.25 ± 0.92 T/S (median 1.14) at baseline and 0.70 ± 0.37 T/S (median 0.70) after 6 years of follow-up. Median 6-year LTL change was -0.46 T/S [interquartile range (IQR) -0.57 to 1.06], equating to -0.078 T/S [IQR(-0.092 to 0.176)] per year. Of note, telomere lengthening occurred in 30.4% of subjects. After adjustment for classical cardiovascular disease (CVD) risk factors (age, gender, smoking, physical activity, alcohol consumption, systolic blood pressure, glucose levels, lipid profile and therapies), TS was associated with incident subclinical carotid vascular damage [hazard ratio (HR) 5.19, 95% confidence interval (CI) 1.20-22.4, P = 0.028]. Finally, subjects in whom LTL shortened over time showed an increased risk of incident CVE, compared to those in whom LTL lengthened (HR 1.69, CI 1.02-2.78, P = 0.041). CONCLUSION: These data indicate that TS is associated with increased risk of subclinical carotid vascular damage and increased incidence of CVEs beyond CVD risk factors in the general population, whereas LTL lengthening is protective.

Lower incidence of macrovascular complications in patients on insulin glargine versus those on basal human insulins: a population-based cohort study in Italy.

BACKGROUND AND AIM: The aim of this study was to compare the use of insulin glargine and intermediate/long-acting human insulin (HI) in relation to the incidence of complications in diabetic patients. METHODS AND RESULTS: A population-based cohort study was conducted using administrative data from four local health authorities in the Abruzzo Region (900,000 inhabitants). Diabetic patients without macrovascular diseases and treated with either intermediate/long-acting HI or glargine were followed for 3-years; the incidence of diabetic (macrovascular, microvascular and metabolic) complications was ascertained by hospital discharge claims and estimated using Cox proportional hazard models. Propensity score (PS) matching was also used to adjust for significant differences in the baseline characteristics between the two groups. RESULTS: Overall, 1921 diabetic patients were included: 744 intermediate/long-acting HI and 1177 glargine users. During the 3-year follow-up, 209 (28.1%) incident events of any diabetic complication occurred in the intermediate/long-acting HI and 159 (13.5%) in the glargine group. After adjustment for covariates, glargine users had an HR (95% CI) of 0.57 (0.44-0.74) for any diabetic complication and HRs of 0.61 (0.44-0.84), 0.58 (0.33-1.04) and 0.35 (0.18-0.70) for macrovascular, microvascular and metabolic complications, respectively, compared to intermediate/long-acting HI users. PS analyses supported these findings. CONCLUSIONS: The use of glargine is associated with a lower risk of macrovascular complications compared with traditional basal insulins. However, limitations inherent to the study design including the short length of observation and the lack of data on metabolic control or diabetes duration, do not allow us to consider this association as a proof of causality.

Different patterns characterize Omega 6 and Omega 3 long chain polyunsaturated fatty acid levels in blood from Italian infants, children, adults and elderly.

Long chain polyunsaturated fatty acids (LC-PUFA), especially the Omega 3, modulate key functions in the body. Their circulating levels are representative of their "status", and may vary at different ages. We have compared the FA status in Italian subjects from neonates to adulthood, assessed through FA analysis of blood drops from fingertips. Data from four cohorts of Italian subjects (total number 1835), have been pooled in four age-groups: neonates (4 days, n=81), children (2-9 years, n=728), adults (40-59 years, n=434) and elderly (60-79 years, n=592). LC-PUFA of both series (Omega 3 and 6) are higher in the blood of neonates than at subsequent ages, reflecting the efficient transfer of these FA from mothers to the fetus. In contrast, the lowest levels of Omega 3 PUFA, especially of DHA, are found in children, probably reflecting inadequate dietary intakes, with possible consequences on the health status at subsequent ages.

How many patients need statin treatment in a low-cardiovascular-risk country? Low-density lipoprotein-cholesterol target and distance from target distribution in an Italian cohort.

BACKGROUND AND AIM: To assess cardiovascular risk distribution, distribution of individual low-density lipoprotein (LDL)-cholesterol target and distance of LDL cholesterol from the target in a representative sample of the Italian population. METHODS AND RESULTS: Cross-sectional, population-based study of a representative sample of the Italian adult population, comprising 5458 individuals (from 40 to 79 years of age, both sexes) from general practices in Italy. Of the subjects, 65.2% were in the low-cardiovascular-risk class, whereas 10.5%, 18.3% and 6.0% had moderate, high, and very high cardiovascular risk profiles, respectively; 8.2% of the subjects were treated with statins at enrolment. Of the cohort, 68.3% displayed LDL-cholesterol values below their LDL target, as calculated according to their individual risk profile. Among the 31.7% 'not at target', 42.3% were ≤ 15%, 44.3% were between 15% and 40% and 13.4% were >40% over their LDL target. CONCLUSIONS: About two-thirds of adults in a low-cardiovascular-risk country, such as Italy, have LDL-cholesterol levels 'at target', as defined in current guidelines. Accordingly, the remaining subjects require a lifestyle or pharmacological intervention to reach their target; 24% of the total cohort, in detail, need to be treated with a statin (or to continue the prescribed statin treatment) to reach the proper LDL target. This type of data analysis might help to optimise resource allocation in preventive medicine.

Low density lipoprotein oxidation, antioxidants, and atherosclerosis.

Oxidized low density lipoproteins (LDLs) are believed to be the most atherogenic form of LDL. However, although a number of experimental data support this concept, the protective role of antioxidants that may prevent LDL oxidation in atherosclerosis is only partially confirmed by studies in humans. Observational and epidemiologic data as well as randomized trials failed to provide clear-cut indications because of mixed results on the protective role of antioxidants against cardiovascular diseases. In spite of the lack of a general consensus, recent data reinforce the concept that a regular intake of antioxidants present in food blocks the progression of atherosclerosis and that the reduced oxidisability of LDL may represent a good marker to follow the action of antioxidants. When it becomes possible to monitor the efficacy of any antioxidant therapy with validated markers of oxidation, the potential influence of vitamins and antioxidants on coronary artery disease will eventually be resolved.

Antioxidants and coronary artery disease.

The protective role of antioxidants in experimental models of atherosclerosis is only partially confirmed by studies in man. Observational and epidemiologic data, as well as randomized trials failed to provide clear cut indications, because of mixed results on the protective role of antioxidants against cardiovascular diseases. In spite of the lack of a general consensus, recent data reinforce the concept that a regular intake of antioxidants present in food limits the progression of atherosclerosis. When it will be possible to monitor the efficacy of any antioxidant therapy with validated markers of oxidation, the potential influence of vitamins and antioxidants on coronary artery disease may eventually be resolved.

Identification of [14C]carmoisine metabolites in bacterial suspension of rat faeces.

An in vitro system consisting of a bacterial suspension of human or rat faecal microflora brought about the biological reduction of the red azo dye [14C]carmoisine to 1-naphthyl-amine-4-sulphonic acid (NA) and 2-amino-1-naphthol-4-sulphonic acid (ANA). These metabolites have been unequivocally identified by radio-HPLC, spectroscopic methods, dilution with cold authentic standards and evidence that the specific activity of the diluted compounds remained constant throughout repeated crystallization, acetylation and purification. The results clearly indicated that samples derived from anaerobic incubations have to be processed for analysis in the complete absence of oxygen. In the presence of oxygen, the formation of a complex pattern of compounds in addition to NA was observed as a consequence of the chemical decomposition of ANA.

Anti-inflammatory activity of Echinacea angustifolia fractions separated on the basis of molecular weight.

Five fractions of an aqueous extract obtained from the roots of Echinacea angustifolia were separated on the basis of molecular weight. The topical anti-inflammatory activity of the fractions has been evaluated in mice using the Croton oil ear test. The fraction with a molecular weight between 30,000 and 100,000 was the most active in inhibiting the oedema; it also reduced the infiltration of inflammatory cells. The activity of this fraction was comparable with that of a raw polysaccharidic extract obtained from E. angustifolia roots by differential solubility. The high-molecular weight polysaccharides are therefore proposed as the anti-inflammatory principles of the plant.

Reduction rate of 14C-Carmoisine by resting cell bacterial suspension from human and rat faeces.

14C-Carmoisine (250/ug; 1.25 x 10(6) dpm) was incubated under strictly anaerobic conditions with resting cell suspension from stool specimens collected from male rats and human male healthy adults. The kinetics of azoreduction was determined as amount of naphthionic acid (NA), the stable metabolite of Carmoisine produced by the activity of the anaerobic bacteria. The analytical determinations were performed by radio-HPLC technique. There were no significant qualitative differences in the radiochromatographic profiles of samples obtained from human and rat flora suspensions. The calculated reduction rates were 5.03 +/- 0.18 nmoles of NA/250/ug protein/min, and 1.72 +/- 0.12 nmoles of NA/250/ug protein/min for rat and human faecal resting cells respectively. From our results it seems that the enzymatic reaction, following zero order kinetics, is similar for the two species and only the reduction rate is different.

Arachidonic acid metabolism in HEL/30 murine epidermal cell line.

The established mouse epidermis-derived cell line HEL/30 was incubated in the presence of 3H arachidonic acid (AA) for 1 h. After medium removal, cells were reincubated with fresh medium in the presence or absence of the calcium ionophore A23187 and tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The AA metabolites formed were extracted from cell-free medium and analyzed using TLC and HPLC. The distribution of the recovered radioactivity showed PGE2, 15-hydroxy-eicosatetraenoic acid (15-HETE), and leukotriene B4 (LTB4), as major products of AA metabolism. The presence of calcium ionophore A23187 increased the release of radioactivity, without affecting the profile of metabolites present in the medium. TPA elicited a preferential increase of cycloxygenase metabolism, this effect being reversed by indomethacin. 5,8,11,14-eicosatetraynoic acid (ETYA) almost completely inhibited LT and HETE formation in A23187 and TPA-treated cells. The results show that HEL/30 cells are able to metabolize AA via both cyclo- and lipoxygenase pathways and that these activities can be modified by chemical means. This cell line might be a suitable tool for studying the involvement of arachidonic acid cascade in cell response to exogenous stimuli.

Anti-inflammatory activity of a polysaccharidic fraction of Echinacea angustifolia.

The anti-inflammatory activity of a polysaccharidic fraction (EPF) obtained from Echinacea angustifolia roots has been examined using the carrageenan paw oedema and the croton oil ear test. EPF (0.5 mg kg-1 i.v.) almost inhibited the carrageenan-induced oedema over 8 h and furthermore, EPF, topically applied, inhibited mouse ear oedema induced by croton oil. EPF also reduced the leukocytic infiltration of the croton oil dermatitis, evaluated both as peroxidase activity and histologically. After topical application EPF appears to be slightly inferior in potency to indomethacin. The results suggest that the anti-inflammatory activity of E. angustifolia resides in its polysaccharidic content.

Lack of correlation between TPA-induced prostaglandin biosynthesis and ornithine decarboxylase activity in Balb/c mouse 3T3 fibroblasts.

12-O-tetradecanoylphorbol-13-acetate (TPA) induced in Balb/c 3T3 cells an earliest prostaglandin biosynthesis and an ornithine decarboxylase activation, this time-relation being more evident if serum was added to incubation medium in low concentration (0.2%). However the two TPA-induced events can be almost totally dissociated by pharmacological means, such as indomethacin and calcium-ionophore A23187 which affected PG response to TPA, but did not influence ODC induction.

Similar profile of urinary and faecal metabolites of 14C-carmoisine in male and pregnant female rats after oral administration.

Pregnant rats received 14C-Carmoisine (200 mg kg-1; 25 microCi) by gavage on days 16-19 of gestation. The animals were killed and maternal tissues, amniotic fluid, placentae, foetal membranes and foetuses were analyzed for radioactivity. No evidence for the transplacental transfer of 14C-Carmoisine or its metabolites was obtained. Male rats were given a single oral administration of 14C-Carmoisine (200 mg kg-1; 25 microCi) and killed at different times after dosing. In both male and female animals, more than 90% of the radioactivity was excreted in faeces and urine within 64 h, and the results suggested that there was no significant absorption of the azodye and no preferential concentration of the red food colour or its metabolites in any particular tissue. Analyses by HPLC, combined with a radioactivity monitor (RAM), of urine and faeces of such animals show that five radioactive peaks were present in the radiochromatogram in addition to unmodified Carmoisine. The mean peak shows the retention time and the u.v. spectrum of authentic naphthionic acid. The results demonstrate that the pregnancy does not affect the kinetic and the metabolic profile of a single oral administration of the azodye Carmoisine given at different days of gestation.

Evidence from two classic irritation tests for an anti-inflammatory action of a natural extract, Echinacina B.

The roots of Echinacea angustifolia (fam. Compositae) were used to obtain an antiphlogistic, immunostimulating and skin repairing extract. On the basis of these potential actions, the extract is used in cosmetic preparations. The aim of this study was to evaluate the anti-inflammatory activity of the extract using different irritation tests. The irritation reaction was induced by application of 0.015 ml of 0.25% croton oil in water to the ears of mice. The raw extract (Echinacina B), topically applied, inhibited oedema both at the maximum (6 hr) and in the decreasing phase (18 hr), and this effect was directly proportional to the doses used. Echinacina B was found to be more potent than the positive control, benzidamine, a topical non-steroidal anti-inflammatory drug. In addition, the extract given iv 1 hr before injection of 0.05 ml of 1% carrageenan into the hind paws of rats inhibited oedema in the histaminic and in the later phases of the phlogistic process. These data show the qualitative value of irritation tests for studying the anti-inflammatory action of a natural plant extract.

Quantitative analysis of alpha, beta-thujone, pulegone, safrole, coumarin and beta-asarone in alcoholic beverages by selected-ion monitoring.

Interest in the quantitative analysis of flavouring agents in alcoholic beverages arises from the possible toxicological consequences deriving from their use. The method reported here, based on gas chromatography with selected-ion monitoring mass spectrometry (SIM), allows the simultaneous measurement of alpha, beta-thujone, pulegone, safrole, coumarin and beta-asarone at the ppb level in alcoholic beverages.

Application of a radioactivity detector to the analysis of 14C-carmoisine metabolites by ion-pair high-pressure liquid chromatography.

14C-Carmoisine was incubated under anaerobic conditions with a suspension of human faeces. Analyses of the incubation medium by high-performance liquid chromatography (HPLC) attached to a radioactivity monitor (RAM) showed the same radioactivity profile as the urine and faeces of rats dosed with the same azodye (200 mg kg-1; 25 microCi). The analyses were carried out with a 5 micron RP-C18 chromatographic column, using a linear gradient profile of different concentrations of water, methanol and an ion-pair reagent. Five radioactive peaks were present in the radiochromatogram , in addition to unmodified Carmoisine. The major peak retained half of the specific activity of Carmoisine, and exhibited the retention time and the u.v. spectrum of authentic naphthionic acid. The results demonstrate the value and the advantage of using the in vitro preparation as a model to detect and to identify the metabolites of similar synthetic azodyes used as food additives.