Francisco Azzato

Chronic cola drinking induces metabolic and cardiac alterations in rats

AIM To investigate the effects of chronic drinking of cola beverages on metabolic and echocardiographic parameters in rats. METHODS Forty-eight male Wistar rats were divided in 3 groups and allowed to drink regular cola (C), diet cola (L), or tap water (W) ad libitum during 6 mo. After this period, 50% of the animals in each group were euthanized. The remaining rats drank tap water ad libitum for an additional 6 mo and were then sacrificed. Rat weight, food, and beverage consumption were measured regularly. Biochemical, echocardiographic and systolic blood pressure data were obtained at baseline, and at 6 mo (treatment) and 12 mo (washout). A complete histopathology study was performed after sacrifice. RESULTS After 6 mo, C rats had increased body weight (+7%, P < 0.01), increased liquid consumption (+69%, P < 0.001), and decreased food intake (-31%, P < 0.001). C rats showed mild hyperglycemia and hypertriglyceridemia. Normoglycemia (+69%, P < 0.01) and sustained hypertriglyceridemia (+69%, P < 0.01) were observed in C after washout. Both cola beverages induced an increase in left ventricular diastolic diameter (C: +9%, L: +7%, P < 0.05 vs W) and volumes (diastolic C: +26%, L: +22%, P < 0.01 vs W; systolic C: +24%, L: +24%, P < 0.05 vs W) and reduction of relative posterior wall thickness (C: -8%, L: -10%, P < 0.05 vs W). Cardiac output tended to increase (C: +25%, P < 0.05 vs W; L: +17%, not significant vs W). Heart rate was not affected. Pathology findings were scarce, related to aging rather than treatment. CONCLUSION This experimental model may prove useful to investigate the consequences of high consumption of soft drinks.

Morning surge, pulse wave velocity, and autonomic function tests in elderly adults.

ObjectiveTo assess the complex interplay between morning surge (MS), the autonomic reflex response at the cardiovascular level, and target organ damage (arterial stiffening, left ventricle hypertrophy). MethodsFifty-nine consecutive elder patients (>65 years old) underwent a 24-h ambulatory blood pressure monitoring. Pulse wave velocity (PWV) was measured as an indicator of arterial stiffness. Autonomic status was assessed by scoring five conventional tests [handgrip, orthostatic pressor response, Valsalva maneuver, heart rate variation during deep breathing (‘I:E’), and immediate heart rate response to standing (‘30 : 15’)]. Results(a) MS was correlated to left ventricle mass (P<0.005), the orthostatic pressor response (P<0.02), and blood pressure variability (BPVar) (P<0.0001) (n=59). (b) PWV explained 61.4% of MS variation for MS values 40 mmHg or less (84% of patients) (P<0.03, n=49) and 38% of MS variation in nondippers (P<0.04, n=25). (c) There were sex-related differences. PWV was associated with the orthostatic pressor response (P<0.02), ‘I:E’ values (P<0.04) and the ‘30 : 15’ test (P<0.04) in men (n=14). In women (n=41), the ‘I:E’ values were associated with MS and BPVar (P<0.003). ConclusionMS was closely related to PWV (arterial stiffening) and BPVar in a small urban sample of cardiovascular patients. MS was also associated with dysautonomia (orthostatic blood pressure/heart rate response to challenges), mostly with impaired parasympathetic modulation. MS and high BPVar cause left ventricular hypertrophy, whereas arterial stiffness alters baroreceptor sensitivity, which in turn affects BPVar, perpetuating a vicious cycle. These findings, although obtained in a small number of participants, provide relevant information not yet available in the local databases.

Histomorphometry of Umbilical Cord Blood Vessels in Preeclampsia

The authors hypothesized that preeclampsia may change the phenotype of umbilical cord vessels. Segments of umbilical cords were obtained from 29 pregnant women (20 healthy and 9 with preeclampsia), which were histomorphometrically assessed. Birth weight was 2928±613 g for the control group vs 1749±656 g for the preeclampsia group (P<.0001). A significantly shorter gestational period was noted in the preeclampsia group: 35 weeks vs 39 weeks in the healthy group. Measurements of the outer layer area (116.4±55 μm2 vs 56.5±25 μm2; P=.0038), the inner layer area (63.1±16 μm2 vs 28.6±8 μm2; P<.0001), the lumen area (8.4±1 μm2 vs 3.4±2 μm2; P=.0003), and the wall/lumen ratio (20.3±9 vs 3.1±0.6; P<.0001) of arteries were significantly larger in the preeclampsia umbilical cords. Concerning veins, the wall/lumen ratio was higher in the preeclampsia group. In this study, the umbilical cord in preeclampsia showed significant changes in the structure of umbilical arteries, with increases in wall areas and wall/lumen ratios. J Clin Hypertens (Greenwich). 2011;13:30–34. ©2010 Wiley Periodicals, Inc.

Coronary Intimal Thickening in Newborn Babies and ≤1-Year-Old Infants

We performed a morphological characterization of intimal thickenings in coronary arteries in the very early stages of life to obtain insights into initial coronary atherogenesis. We examined specimens from 67 infants who had died of noncardiac causes within their first year of life. Serially cut sections were stained with hematoxylin-eosin, Azan, Alcian blue, acetic orceine, and immunotypified for CD68, CD34, and α-smooth muscle (SM) actin. Substantial changes were detected in about 1 of 3 participants. Alterations ranged from focal areas with mild myointimal thickening to diffuse moderate thickening. In those lesions, smooth muscle cells (SMCs) showed loss of polarity, infiltrating the subendothelium, mostly with rupture of the internal elastic lamina and without neoangiogenesis. Morphometrically, in musculoelastic intimal thickenings, neointimal thickness averaged 58.3 ± 17.8 µm, affecting 46% of the internal elastic membrane perimeter; lumen stenosis averaged 13.7% ± 5.0%. These lesions can be present very early in life and SMCs seem to play an essential role.

Carotid barochemoreceptor pathological findings regarding carotid plaque status and aging

BACKGROUND Carotid barochemoreceptor pathological lesions have been studied in animals, but few human necropsies have been performed. Therefore, data rely on case patients following surgery, radiotherapy and carotid endarterectomy. Almost no data are available regarding whether the effect of aging prevails over pathological conditions, despite the classic description that glomic fibrosis increases with age. OBJECTIVE To morphometrically characterize the alterations of the carotid barochemoreceptors and their supplying arteries. METHODS Patients (n=23) who had suffered and died from stroke, with and without complicated internal carotid atheromatosis, were divided by age (group 1: older than 80 years; group 2: 65 to 80 years; and group 3: younger than 65 years). Carotid segments were obtained at autopsy. The specimens were stained for light microscopy and immunohistochemistry. RESULTS Carotid glomus presented from moderate-to-severe atrophy and fibrosis. A focal decrease in vascularization (CD34-positive) of the glomus (greater than 50%) was observed in areas of atrophy and fibrosis. Damaged nerve endings (S100 protein-positive) were observed at the media of the carotid sinus. Morphometric data showed no differences between groups for glomus area, number of type 1 and 2 cells, and the wall to lumen arteriole ratio. No statistical differences were demonstrated in the pathological findings of the carotid glomus when comparing complicated with noncomplicated plaques or age groups. CONCLUSION Severe carotid chemoreceptor damage exists in patients who have died from stroke and suffered from carotid atheromatosis. These findings were independent from aging and plaque type. However, damage was correlated with a marked narrowing of the supplying arterioles as a consequence of hemodynamic and/or metabolic alterations (dyslipidemia, diabetes).

Cardiorenal Involvement in Metabolic Syndrome Induced by Cola Drinking in Rats: Proinflammatory Cytokines and Impaired Antioxidative Protection.

We report experimental evidence confirming renal histopathology, proinflammatory mediators, and oxidative metabolism induced by cola drinking. Male Wistar rats drank ad libitum regular cola (C, n = 12) or tap water (W, n = 12). Measures. Body weight, nutritional data, plasma glucose, cholesterol fractions, TG, urea, creatinine, coenzyme Q10, SBP, and echocardiograms (0 mo and 6 mo). At 6 months euthanasia was performed. Kidneys were processed for histopathology and immunohistochemistry (semiquantitative). Compared with W, C rats showed (I) overweight (+8%, p < 0.05), hyperglycemia (+11%, p < 0.05), hypertriglyceridemia (2-fold, p < 0.001), higher AIP (2-fold, p < 0.01), and lower Q10 level (−55%, p < 0.05); (II) increased LV diastolic diameter (+9%, p < 0.05) and volume (systolic +24%, p < 0.05), posterior wall thinning (−8%, p < 0.05), and larger cardiac output (+24%, p < 0.05); (III) glomerulosclerosis (+21%, p < 0.05), histopathology (+13%, p < 0.05), higher tubular expression of IL-6 (7-fold, p < 0.001), and TNFα (4-fold, p < 0.001). (IV) Correlations were found for LV dimensions with IL-6 (74%, p < 0.001) and TNFα (52%, p < 0.001) and fully abolished after TG and Q10 control. Chronic cola drinking induced cardiac remodeling associated with increase in proinflammatory cytokines and renal damage. Hypertriglyceridemia and oxidative stress were key factors. Hypertriglyceridemic lipotoxicity in the context of defective antioxidant/anti-inflammatory protection due to low Q10 level might play a key role in cardiorenal disorder induced by chronic cola drinking in rats.

Antioxidants Supplementation in Elderly Cardiovascular Patients

Supplementation with antioxidants and its benefit-risk relationship have been largely discussed in the elderly population. We evaluated whether antioxidants supplementation improved the biochemical profile associated with oxidative metabolism in elderly cardiovascular patients. Patients (n = 112) received daily supplementation with α-TP 400 mg, beta-carotene 40 mg, and vitamin C 1000 mg for 2 months (treatment). Plasma concentrations of alpha-tocopherol (α-TP), β-carotene (βC), ubiquinol-10 (QH-10), glutathione, and thiobarbituric acid reactive substances (TBARS) were determined before and after treatment. Response to treatment was dependent on pretreatment α-TP and βC levels. Increase in α-TP and βC levels was observed only in patients with basal levels <18 μM for α-TP (P < 0.01) and <0.30 μM for βC (P < 0.02). Ubiquinol-10, glutathione, and TBARS were unaffected by treatment: QH-10 (+57%, F 1,110 = 3.611, P < 0.06, and N.S.), glutathione (+21%, F 1,110 = 2.92, P < 0.09, and N.S.), and TBARS (−29%, F 1,110 = 2.26, P < 0.14, and N.S.). Treatment reduced oxidative metabolism: 5.3% versus 14.6% basal value (F 1,110 = 9.21, P < 0.0003). Basal TBARS/α-TP ratio was higher in smokers compared to nonsmokers: 0.11 ± 0.02 versus 0.06 ± 0.01 (F 32,80 = 1.63, P < 0.04). Response to antioxidant supplementation was dependent on basal plasma levels of α-TP and βC. Smoking status was strongly associated with atherosclerotic cardiovascular disease and high TBARS/α-TP ratio (lipid peroxidation).

A prospective observational cohort study highlights kidney biopsy findings of lupus nephritis patients in remission who flare following withdrawal of maintenance therapy

One of the most difficult management issues in lupus nephritis (LN) is the optimal duration of maintenance immunosuppression after patients are in clinical remission. Most patients receive immunosuppression for years, based mainly on expert opinion. Prospective data are unavailable. Complicating this issue are data that patients in clinical remission can still have histologically active LN; however, the implications of this are unknown. To study this, the Lupus Flares and Histological Renal Activity at the end of Treatment study (ClinicalTrial.gov, NCT02313974) was designed to examine whether residual histologic activity predisposes to LN flares in class III and IV LN. Patients in complete clinical remission for at least 12 months who had received at least 36 months of immunosuppression were eligible. Patients consented to a second kidney biopsy, were tapered off maintenance immunosuppression and were then followed prospectively for LN flares over 24 months. Forty-four patients were enrolled, and 36 completed the study. LN flares occurred in 11 patients, and ten of these had residual histologic activity on the second biopsy. All patients with an NIH activity index over two flared. The activity index and duration of systemic lupus erythematosus at the second biopsy were independent predictors of flare. A predictive equation based on these variables discriminated between flare and no flare with a sensitivity of 100%, specificity of 88%, and a misclassification rate of 8.3%. Thus, a repeat kidney biopsy may be useful in managing maintenance immunosuppression in LN, and patients in histologic remission may be candidates for withdrawal of therapy.

Ozonetherapy protects from in-stent coronary neointimal proliferation. Role of redoxins.

BACKGROUND In-stent restenosis and poor re-endothelization usually follow percutaneous transluminal coronary angioplasty, even using drug-eluting stents, due to inflammation and oxidative stress. Medical ozone has antioxidant and anti-inflammatory properties and has not been evaluated in this context. OBJECTIVES To evaluate whether ozonotherapy might reduce restenosis following bare metal stents implantation in relation to the redoxin system in pigs. METHODS Twelve male Landrace pigs (51±9kg) underwent percutaneous transluminal circumflex coronary arteries bare metal stent implantation under heparine infusion and fluoroscopical guidance, using standard techniques. Pigs were randomized to ozonetherapy (n=6) or placebo (n=6) treatment. Before stenting (24h) and twice a week for 30days post-stenting, venous blood was collected, ozonized and reinfused. Same procedure was performed in placebo group except for ozonation. Both groups received antiplatelet treatment. Histopathology and immunohistochemistry studies were performed. RESULTS Severe inflammatory reaction and restenosis with increase in the immunohistochemical expression of thioredoxin-1 were observed in placebo group 30days after surgery. Oppositely, ozonetherapy drastically reduced inflammatory reaction and restenosis, and showed no increase in the Trx-1 immunohistochemical expression 30days after surgery. Immunolabeling for Prx-2 was negative in both groups. Ozonated autohemotherapy strikingly reduced restenosis 30days following PTCA with BMS implantation in pigs. CONCLUSIONS Stimulation of the redoxin system by ozone pretreatment might neutralize oxidative damage from the start and increase antioxidative buffering capacity post-injury, reducing further damage and so the demand for antioxidant enzymes. Our interpretation agrees with the ozone oxidative preconditioning mechanism, extensively investigated.

Exercise Ameliorates Endocrine Pancreas Damage Induced by Chronic Cola Drinking in Rats.

Purpose This study evaluates whether the daily practice of an exercise routine might protect from endocrine pancreas damage in cola drinking rats. Methods Forty-eight Wistar rats were randomly assigned to 4 groups depending on a) beverage consumption ad libitum, water (W) or cola beverage (C), and b) physical activity, sedentary (S) or treadmill running (R). Accordingly, 4 groups were studied: WS (water sedentary), WR (water runner), CS (cola sedentary) and CR (cola runner). Body weight, nutritional data, plasma levels of glucose, creatinine, total cholesterol and cholesterol fractions, and triglycerides (enzymocolorimetry), and systolic blood pressure (plethysmography) were measured. After 6 months, euthanasia was performed (overdose sodium thiopental). Pancreatic tissue was immediately excised and conventionally processed for morphometrical and immunohistochemical determinations. Results The effects of running and chronic cola drinking on pancreas morphology showed interaction (p<0.001) rather than simple summation. Cola drinking (CS vs WS) reduced median pancreatic islet area (-30%, 1.8 104 μm2 vs 2.58 104 μm2, p<0.0001) and median β-cell mass (-43%, 3.81 mg vs 6.73 mg, p<0.0001), and increased median α/β ratio (+49%, 0.64 vs 0.43, p< 0.001). In water drinking rats (WR vs WS), running reduced median α-cell mass (-48%, 1.48 mg vs 2.82 mg, p<0.001) and α/β ratio (-56%, 0.19 vs 0.43, p<0.0001). Differently, in cola drinking rats (CR vs CS), running partially restored median islet area (+15%, 2.06 104 μm2 vs 1.79 104 μm2, p<0.05), increased median β-cell mass (+47%, 5.59 mg vs 3.81 mg, p <0.0001) and reduced median α/β ratio (-6%, 0.60 vs 0.64, p<0.05). Conclusion This study is likely the first reporting experimental evidence of the beneficial effect of exercise on pancreatic morphology in cola-drinking rats. Presently, the increase of nearly 50% in β cells mass by running in cola drinking rats is by far the most relevant finding. Moderate running, advisably indicated in cola consumers and patients at risk of diabetes, finds here experimental support.