Effects of pioglitazone versus metformin on circulating endothelial microparticles and progenitor cells in patients with newly diagnosed type 2 diabetes--a randomized controlled trial.

AIM: Endothelial microparticles (EMPs) and endothelial progenitor cells (EPCs) are markers of endothelial injury and repair. We compared the effects of pioglitazone versus metformin on the circulating numbers of EMPs and EPCs in patients with newly diagnosed type 2 diabetes. METHODS: This was a randomized, double-blind, comparator-controlled, 24-week single-centre trial conducted in a Teaching Hospital in Naples, Italy. One hundred and ten people with newly diagnosed type 2 diabetes who were never treated with antihyperglycaemic drugs and had haemoglobin A1c (HbA1c) levels between 7 and 10% were given pioglitazone hydrochloride (15-45 mg/day) (n = 55) or metformin (1000-2000 mg/day) (n = 55) as an active comparator. Absolute change from baseline to final visit in circulating EMPs and EPCs and their ratio were the main outcomes. RESULTS: Baseline characteristics did not differ between the study groups. The decrease in circulating EMPs CD31+ [intergroup difference, -32 counts/µl (95% CI -51 to -9)] and the increase in EPCs CD34+/KDR+ [intergroup difference, 33 cells/10(6) events (95% CI 13 to 55)] were greater with pioglitazone versus metformin. EMPs/EPCs ratio was reduced with pioglitazone and unchanged with metformin [difference, -1.5 (95% CI -2.6 to -0.5), p < 0.001]. Participants assigned to pioglitazone gained more weight and experienced greater improvements in some coronary risk measures [high-density lipoprotein (HDL)-cholesterol, triglycerides, adiponectin and C-reactive protein (CRP)] than did those assigned to metformin. CONCLUSION: Compared with metformin, pioglitazone treatment improved the imbalance between endothelial damage and repair capacity and led to more favourable changes in coronary risk factors in patients with newly diagnosed type 2 diabetes.

Interleukin-20 circulating levels in obese women: effect of weight loss.

BACKGROUND AND AIMS: Obesity is associated with an increased risk of developing atherosclerosis. Interleukin-20 (IL-20) is a pleiotropic cytokine thought to be involved in the onset and progression of atherosclerosis. The aim of this study was to determine whether circulating levels of IL-20 are elevated in obese women and whether they could be affected by a substantial decrease in body weight. METHODS AND RESULTS: Fifty obese and 50 age-matched, normal weight, premenopausal women participated in the study. Obese women entered into a medically supervised weight loss program aimed at reducing body weight to 90% of baseline. We measured anthropometric, glucose and lipid parameters, and IL-20, C-Reactive Protein (CRP) and interleukin-10 (IL-10) circulating levels. Circulating IL-20 and CRP levels were significantly higher in obese than control women (P=0.01), while IL-10 levels were significantly lower; IL-20 levels were positively associated with body weight (r=0.35; P=0.02) and visceral fat (waist-hip ratio; r=0.32; P=0.025). Caloric restriction-induced weight loss (>10% of original weight) over 6 months reduced IL-20 levels from 152 (112/184) to 134 (125/153)pg/ml (median and 25%/75%; P=0.03), and it was positively associated with changes in body mass index and waist-hip ratio. CONCLUSION: In premenopausal obese women, IL-20 levels are higher than matched normal weight control women, are associated with body weight and waist-hip ratio, and are reduced by weight loss.

Adherence to a Mediterranean diet and glycaemic control in Type 2 diabetes mellitus.

AIMS: Mediterranean-type diets reduce the risk of Type 2 diabetes. Whether a Mediterranean-type diet improves glycaemic control in diabetes remains unknown. METHODS: We conducted a cross-sectional analysis in 901 outpatients with Type 2 diabetes attending diabetes clinics located in Campania County, South Italy. We explored the relation between glycated haemoglobin (HbA(1c)), measured centrally, self-measured pre- and postprandial glucose levels and consumption of a Mediterranean-type diet. Adherence to a Mediterranean-type diet was assessed by a 9-point scale that incorporated the salient characteristics of this diet (range of scores, 0-9, with higher scores indicating greater adherence). The study was conducted from 2001 to 2007. RESULTS: Diabetic patients with the highest scores (6-9) had lower body mass index and waist circumferences, a lower prevalence of the metabolic syndrome and lower HbA(1c) and post-meal glucose levels than diabetic patients with the lowest scores (0-3). In multivariate analysis, mean HbA(1c) and 2-h post-meal glucose concentrations were significantly lower in diabetic patients with high adherence to a Mediterranean-type diet than those with low adherence [difference: HbA(1c) 0.9%, 95% confidence intervals (CI) 0.5-1.2%, P < 0.001; 2-h glucose 2.2 mmol/l, 95% CI 0.8-2.9 mmol/l, P < 0.001]. CONCLUSIONS: In Type 2 diabetes, greater adherence to a Mediterranean-type diet is associated with lower HbA(1c) and postprandial glucose levels.

First Report of Fusarium Wilt of Flue-Cured Tobacco Caused by Fusarium oxysporum f. sp. batatas in Spain.

Tobacco (Nicotiana tabaccum L.) is among the most important crops in La Vera Region (Cáceres, west-central Spain) where, since 2001, flue-cured tobacco plants showing typical symptoms of a vascular disease associated to the cyst nematode (Globodera tabacum) complex have been frequently observed (2). Symptoms observed were wilting, yellowing, and drying of the leaves, usually on one side of the plant; stunted and distorted leaves with curved midribs due to the unequal growth; and brown vascular discoloration of stems and midribs. Several diseased plants were collected during the summers of 2003 and 2004, and F. oxysporum was consistently isolated from sections of necrotic midribs. To confirm the pathogenicity of F. oxysporum isolates, plants of flue-cured tobacco (cv. Ct-681) were inoculated. Since Fusarium wilt of tobacco may be caused by F. oxysporum f. sp. vasinfectum or F. oxysporum f. sp. batatas (1), plants of cotton (cv. Acala) and sweet potato (cvs. Nemagold and Nancy Hall) were also inoculated to determine the forma specialis designation. Twenty plants per cultivar and isolate were inoculated with six isolates at the two to four true-leaf stage by pouring into the substrate 200 ml of a conidial suspension (1 × 105 CFU/ml). After inoculation, plants were incubated in a growth chamber at 28°C (day) and 24°C (night) with a 16-h photoperiod. All six F. oxysporum isolates were pathogenic to tobacco and sweet potato, but there were differences among isolates in time from inoculation to appearance of first disease symptoms (7 to 15 days after inoculation in tobacco and sweet potato) and also in disease severity 30 days after inoculation (60 to 100% wilt or mortality in tobacco and 50 to 100% in sweet potato). F. oxysporum was reisolated from stems of inoculated plants. No disease symptom was observed in cotton plants 60 days after inoculation, and F. oxysporum was not reisolated from them. Results of inoculation on the differential hosts indicated that disease symptoms in tobacco were caused by F. oxysporum f. sp. batatas. To our knowledge, this is the first report of F. oxysporum f. sp. batatas causing disease in flue-cured tobacco in Spain. References: (1) G. M. Armstrong and J. K. Armstrong. Phytopathology 58:1242, 1968. (2) G. Espárrago and I. Blanco. Plant Dis. 86:1402, 2002.

Dietary factors in erectile dysfunction.

The role of dietary factors in erectile dysfunction (ED) has never been addressed. In the present case-control study, we investigated the relation of the Mediterranean diet with ED. A total of 100 men with ED were compared with 100 age-matched men without ED. A scale indicating the degree of adherence to the Mediterranean diet was constructed: the total Mediterranean diet score ranged from 0 (minimal adherence to the Mediterranean diet) to 9 (maximal adherence). The percentage of physical inactivity was greater in the ED group (35 vs 19%, P=0.04), whereas the diet score was lower (4.7+/-0.5 vs 5.4+/-0.5, P<0.01), indicating a reduced adherence to the Mediterranean diet. In analyses adjusted for the prevalence of associated risk factors (hypertension, hypercholesterolemia), body mass index, waist, physical inactivity and total energy intake, the intake of fruits and nuts, and the ratio of monounsaturated lipids to saturated lipids remained the only individual measures associated with ED. In conclusion, the results of the present study show that dietary factors may be important in the development of ED: adoption of healthy diets would hopefully help preventing ED.

Erectile dysfunction associates with endothelial dysfunction and raised proinflammatory cytokine levels in obese men.

Erectile and endothelial dysfunction may have some shared pathways through a defect in nitric oxide activity. We evaluated associations between erectile function, endothelial function and markers of systemic vascular inflammation in 80 obese men, aged 35-55 yr, divided into two equal groups according to the presence/absence of erectile dysfunction. Compared with non-obese age-matched men [no.=50, body mass index (BMI)=24 +/- 1], obese men (all) had impaired indices of endothelial function as suggested by the reduced mean blood pressure and platelet aggregation responses to L-arginine, and higher circulating concentrations of the proinflammatory cytokines interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-18 (IL-18), as well as C-reactive protein (CRP). The mean erectile function score was 14 +/- 4 (range 7-19) in obese men with erectile dysfunction and 23.5 +/- 1 (range 22-25) in obese men without erectile dysfunction. Endothelial function showed a greater impairment in impotent obese men as compared with potent obese men. The mean blood pressure and platelet aggregation decreases following L-arginine were -1.5 +/- 1.1 mmHg and -1.1 +/- 1.2%, respectively, in obese men with erectile dysfunction, and -3.4 +/- 1.2 mmHg and -5.6 +/- 2.1%, respectively, in obese men without erectile dysfunction (p < 0.01). Circulating CRP levels were significantly higher in obese men with erectile dysfunction as compared with obese men without erectile dysfunction (p < 0.05). Erectile function score was positively associated with mean blood pressure responses to L-arginine and negatively associated with BMI, waist-to-hip ratio (WHR), and CRR Erectile and endothelial dysfunction associate in obese men and may contribute to their raised cardiovascular risk through impaired nitric oxide availability elicited by a low-grade inflammatory state.

Effect of licorice on the reduction of body fat mass in healthy subjects.

The history of licorice, as a medicinal plant, is very old and has been used in many societies throughout the millennia. The active principle, glycyrrhetinic acid, is responsible for sodium retention and hypertension, which is the most common side-effect. We show an effect of licorice in reducing body fat mass. We studied 15 normal-weight subjects (7 males, age 22-26 yr, and 8 females, age 21-26 yr), who consumed for 2 months 3.5 g a day of a commercial preparation of licorice. Body fat mass (BFM, expressed as percentage of total body weight, by skinfold thickness and by bioelectrical impedance analysis, BIA) and extracellular water (ECW, percentage of total body water, by BIA) were measured. Body mass index (BMI) did not change. ECW increased (males: 41.8+/-2.0 before vs 47.0+/-2.3 after, p<0.001; females: 48.2+/-1.4 before vs 49.4+/-2.1 after, p<0.05). BFM was reduced by licorice: (male: before 12.0+/-2.1 vs after 10.8+/-2.9%, p<0.02; female: before 24.9+/-5.1 vs after 22.1+/-5.4, p<0.02); plasma renin activity (PRA) and aldosterone were suppressed. Licorice was able to reduce body fat mass and to suppress aldosterone, without any change in BMI. Since the subjects were consuming the same amount of calories during the study, we suggest that licorice can reduce fat by inhibiting 11beta-hydroxysteroid dehydrogenase Type 1 at the level of fat cells.

Effects of surgical menopause and estrogen replacement therapy on atrio-ventricular conduction and ventricular repolarization.

OBJECTIVE: To evaluate the effect of surgical menopause and estrogen replacement therapy on atrio-ventricular conduction and ventricular repolarization in women. METHODS: In a prospective, randomized, double-blind, placebo-controlled clinical trial 50 women underwent hystero-salpingo-oophorectomy. Twenty-five women were treated with 50 microg/die of transdermal estradiol and the other 25 were treated with placebo patches. The duration of the treatment was 12 cycles. Before surgery and after 12 cycles of treatment, a standard electrocardiogram was performed to evaluate atrio-ventricular conduction time and cardiac repolarization time. RESULTS: No significant variations in atrio-ventricuar conduction time and cardiac repolarization time were detected between the two groups at entry, nor was there any difference in both groups after 12 cycles of treatment with transdermal estradiol. CONCLUSIONS: Surgical menopause and estrogen replacement therapy do not modify atrio-ventricular conduction and ventricular repolarization in women.

Plasma atrial natriuretic peptide (ANP) fragments proANP (1-30) and proANP (31-67) measurements in chronic heart failure: a useful index for heart transplantation?

The family of the atrial natriuretic peptides, proANP fragments and the active alphaANP, is strongly related to heart disease. The aim was to study in CHF subjects the relation of mdANP and NtANP with brain natriuretic peptide (BNP) and with other traditional medical parameters. Sixteen CHF patients (aged 51.9+/-13.7 years) and 16 healthy subjects age matched (50.8+/-5.9 years) were selected. Both NtANP and mdANP were higher in CHF patients than in healthy subjects (1436+/-288 vs. 288+/-22 pmol/l p<0.001 and 2305+/-383 vs. 423+/-65 pmol/l p<0.0001, respectively). BNP in CHF patients was 28.0+/-9 pmol/l (reference values 1.7+/-1.8 pmol/l). Both NtANP and mdANP demonstrated positive correlation with BNP, p<0.0001 and with left atrial end-systolic volume, p<0.05. BNP correlated with left ventricular mass, p<0.03. In conclusion, plasma NtANP and mdANP analyses are useful laboratory markers in CHF patient investigation and follow up. In particular, they could be employed as non-invasive parameters to follow up worsening of systolic dysfunction until heart transplantation is required.

Plasma lactate, GH and GH-binding protein levels in exercise following BCAA supplementation in athletes.

Branched chain amino acids (BCAA) stimulate protein synthesis, and growth hormone (GH) is a mediator in this process. A pre-exercise BCAA ingestion increases muscle BCAA uptake and use. Therefore after one month of chronic BCAA treatment (0.2 gkg(-1) of body weight), the effects of a pre-exercise oral supplementation of BCAA (9.64 g) on the plasma lactate (La) were examined in triathletes, before and after 60 min of physical exercise (75% of VO2 max). The plasma levels of GH (pGH) and of growth hormone binding protein (pGHBP) were also studied. The end-exercise La of each athlete was higher than basal. Furthermore, after the chronic BCAA treatment, these end-exercise levels were lower than before this treatment (8.6+/-0.8 mmol L(-1) after vs 12.8+/-1.0 mmol L(-1) before treatment; p < 0.05 [mean +/- std. err.]). The end-exercise pGH of each athlete was higher than basal (p < 0.05). Furthermore, after the chronic treatment, this end-exercise pGH was higher (but not significantly, p = 0.08) than before this treatment (12.2+/-2.0 ng mL(-1) before vs 33.8+/-13.6 ngmL(-1) after treatment). The end-exercise pGHBP was higher than basal (p < 0.05); and after the BCAA chronic treatment, this end-exercise pGHBP was 738+/-85 pmol L(-1) before vs 1691+/-555 pmol L(-1) after. pGH/pGHBP ratio was unchanged in each athlete and between the groups, but a tendency to increase was observed at end-exercise. The lower La at the end of an intense muscular exercise may reflect an improvement of BCAA use, due to the BCAA chronic treatment. The chronic BCAA effects on pGH and pGHBP might suggest an improvement of muscle activity through protein synthesis.

Circulating immunoreactive proANP(1-30) and proANP(31-67) in sedentary subjects and athletes.

BACKGROUND: Atrial natriuretic peptide (ANP) is synthesized and stored in myocytes as prohormone(1-126), which upon release is cleaved into proANP(1-98) and alpha-ANP(99-126). In addition, cleavage of proANP(1-98) produces proANP(1-30), proANP(31-67), and proANP(79-98) fragments. ProANP(1-30) and proANP(31-67) have roles in fluid and electrolyte homeostasis. The aim of the present study was to develop a plasma assay for proANP(1-30) and proANP(31-67) and to compare results in trained athletes and sedentary subjects. METHODS: Two competitive enzyme immunoassays were established with affinity-purified sheep antiserum against synthetic ANP fragments. The immunoreactivity (ir) of proANP(1-30) and proANP(31-67) was measured in 10-microL plasma samples without extraction in a microwell-based assay. Plasma concentrations in sedentary male subjects (n = 22) and male endurance athletes (n = 14) were examined. RESULTS: In the assay for ir-proANP(1-30) and ir-proANP(31-67), the concentrations at 95% B/B(0) were 4.7 and 14.2 pmol/L, respectively. Within-run CVs were 4-6% and 5-6%, and between-run CVs were 9% for both assays. Both assays were linear on dilution (y = 0.9945x - 0. 7291 and y = 1.0001x - 3.428), and the recoveries were 102-112% and 102-106%, respectively. In the sedentary and athletic groups, the ir-proANP(1-30) concentrations were similar: 318 +/- 38 pmol/L and 312 +/- 25 pmol/L (mean +/- SE), respectively, whereas the ir-proANP(31-67) was higher in the rowers (713 +/- 81 pmol/L) than in the sedentary subjects (387 +/- 71 pmol/L; P <0.005). CONCLUSIONS: The proANP fragment assays are precise (CV <10%) and exhibit nearly quantitative recovery (102-112%). Only ir-proANP(31-67) responds to physical training.

A rapid urine creatinine assay by capillary zone electrophoresis.

Using capillary zone electrophoresis, the urine creatinine (uCr) assay was validated in extemporaneous diluted urine, both in healthy subjects and athletes, with the uCr concentration as a reference value to compare excretion rates of other metabolites in the same samples. The electrokinetic sample injection was carried out at 10 kV per 10 s; UV absorbance detection was at 254 nm. Using standard samples, the creatinine migration mean time in 100 mmol/L acetate buffer, pH 4.4, was 3.3+/-0.2 min; the repeatability for absolute migration mean time was 0.6% and peak height repeatability was 2.9%. The correlation coefficient of the standard curve was r = 0.999 and the detection limit was 23.1 micromol/L. Intra- and interassay coefficients of variation (CV) were 3.0 and 3.6%, respectively; recovery was 99+/-3% and linearity was r= 0.98. Normal urine samples were diluted 1:80 in run buffer. The present CE urine creatinine assay showed a good correlation with HPLC and with Jaffe methods (r = 0.98 and r = 0.97, respectively; p < 0.0001). The uCr in the morning urine samples of 34 healthy males (M), 38 healthy females (F), and 83 male athletes (A) was 10.4+/-6.1 mmol/L, 10.8+/-8.1 mmol/L and 13.2+/-6.5 mmol/L, respectively. The uCr difference (p < 0.02) between M and A and a correlation (p < 0.05) with age in A were observed.

Regulation of corticosteroid receptors in patients with anorexia nervosa and Cushing's syndrome.

We have studied 16 patients with anorexia nervosa (11 with a stabilised weight loss and 5 in the weight-losing phase), 11 healthy controls, and 10 patients with Cushing's syndrome, by measuring plasma cortisol (by enzyme-immunoassay), ACTH (by RIA), corticosteroid (Type I-mineralocorticoid and Type II-glucocorticoid) receptors in mononuclear leukocytes (by radio-receptor assay), and lymphocyte subpopulations (by cytofluorimetry). In anorexic patients with a stabilised weight loss and in Cushing's syndrome the mean value of both Type I and Type II corticosteroid receptors in mononuclear leukocytes was significantly lower than in controls. The correlation between Type II receptors and plasma cortisol was inverse in stabilised anorexia nervosa and in Cushing's syndrome, and direct in healthy controls. Anorexic patients in the weight-losing phase showed a significant increase in plasma cortisol levels and a normal number of Type II receptors. From these results we hypothesise that in anorexia nervosa there is a progression from an increase in plasma cortisol in the weight-losing phase, to a concomitant decrease in Type II receptors when the disease is stabilised.

Free carnitine and acetyl carnitine plasma levels and their relationship with body muscular mass in athletes.

The purpose of the present study was to investigate the relationship between plasma carnitine concentration and body composition variation in relation to muscular and fat masses since there is no experimentally proved correlation between plasma carnitine and body masses. We used bioelectric impedance analysis (BIA), to determine body composition and to have a complete physical fitness evaluation. The post-absorptive plasma free carnitine and acetyl carnitine plasma levels, body composition as Fat-Free Mass (FFM) and Fat Mass (FM) in kg, as well as in percent of body mass, were analysed in 33 healthy subjects. A significant negative correlation was found between plasma acetyl carnitine and FFM in weight (kg) as well as in percent of body mass (respectively p < 0.0001; p < 0.01); a significant positive correlation was found only between FM in percent and plasma acetyl carnitine (p < 0.01). The observed negative correlation between plasma acetyl carnitine and muscular mass variation might reflect an oxidative metabolic muscle improvement in relation to muscular fat free mass increment and might be evidence that muscle metabolism change is in relation to plasma acetyl carnitine concentration.

Branched-chainα-amino acid chronic treatment: responses of plasmaα-keto-related compounds and ammonia when used in physical exercise performance.

To examine the effects of acute branched-chainα-amino acids (BCAA) oral administration following chronic BCAA intake, a group of well trained young swimmers (n = 12) was submitted to a one month chronic BCAA treatment (0.2g/Kg body weight per die; Leu: Val: Ileu = 2:1:1) and a physical exercise test before and after this period of treatment was carried out. The exercise tests (60min swim) were performed in a high circulating BCAA level state which was obtained through oral BCAA administration (or placebo) just before the beginning of the exercise. The groups will be referred to as BCAA/before, BCAA/after, placebo/before, placebo/after. Blood and plasma (antecubital vein) samples were collected from the different groups at different times: on the morning of the day before the test (basal time, rest 0), the following day 30min after an acute administration (oral dose placebo or BCAA acute treatment: Leu 4.8g, Val 2.4g, Ileu 2.4g), just before the beginning of the exercise performance (time 0min, rest 1), at the end of the exercise (time 60min, EE) and during recovery (time 120min, Re). Plasma ammonia levels increased significantly from rest 1 to the end of the exercise in all subjects, but it was significantly higher in BCAA treated than in placebo subjects in both the before and after chronic treatment groups (BCAA/before: from 38 ± 7 to 204 ± 65mmol/l; placebo/before: from 36 ± 10 to 93 ± 29mmol/l; BCAA/after: from 36 ± 9 to 171 ± 43mmol/l; placebo/after: from 30 ± 6 to 65 ± 16mmol/l). Plasma ammonia level increments observed before a chronic one month BCAA treatment were significantly higher than after this treatment (p < 0.05). Plasma alanine was at all times of the test higher before the BCAA chronic treatment than after; this difference resulted significant at rest 0, rest 1 and recovery times (p < 0.05). After acute BCAA administration, plasma BCAA levels increased from 618 ± 52mmol/l to 1893 ± 284mmol/l (p < 0.05) from the onset of exercise and remained elevated throughout the test. Placebo and basal (rest 0) levels both before and after the chronic treatment did not demonstrate any significant differences. Plasma BCAA and BCKA levels, in the BCAA/before demonstrated significantly higher levels than placebo/before at rest 1 time (BCAA/before vs placebo/before: Leu 86 ± 27 vs 620 ± 97mmol/l; KIC 60 ± 3 vs 87 ± 5mmol/l, Ileu 51 ± 19 vs 359 ± 56mmol/l, KMV 26 ± 1 vs 43 ± 2mmol/l, Val 290 ± 79 vs 915 ± 133mmol/l, KIV 14 ± 1 vs 24 ± 2mmol/l). The levels after the chronic treatment maintained circa these differences in the two groups BCAA/after and placebo/after. The plasma BCAA as well as the BCKA levels of acutely treated athletes, in physical exercise, showed a different profile before and after the chronic treatment. The chronic treated BCAA/after group in fact depicted a decreasing BCKA level profile at the end of the exercise and during recovery; on the contrary, before the chronic treatments, acutely treated athletes demonstrated a tendency to increase these levels during recovery. These data seem to confirm that increased BCAA availability, before exercise, result in significantly greater plasma ammonia responses during exercise than does placebo administration; furthermore this increment is lower after chronic treatment. The interpretation of the ammonia data is difficult since the exercise type could have an influence on this phenomenon. The differences in the profile patterns of alanine, BCAA and BCKA levels seem to indicate that the chronic treatment brings about a state in which there is a better use of BCAA compounds as energy supply.

Effect of naltrexone treatment on insulin secretion, insulin action and postprandial thermogenesis in obesity.

For many years a series of studies has been carried out to evaluate the role of endogenous opioid peptides on glucose metabolism. In this work we studied the influence of endogenous opioid peptides on insulin response to OGTT and glucose-induced thermogenesis before and after a prolonged oral treatment with Naltrexone (50 mg/daily for 6 days), an opioid receptor antagonist, in a group of 9 obese subjects. Moreover in obese patients we evaluated the effect of this anti-opioid drug on insulin secretion and insulin sensitivity during an IVGTT using the minimal model approach. We compared the pre-treatment results with data coming from a group of 5 normal-weight subjects. We measured blood glucose, plasma insulin and C-peptide concentrations and evaluated the following parameters: first (phi 1) and second (phi 2) phase of beta-cell sensitivity, insulin sensitivity and glucose effectiveness. Obese subjects displayed an increased insulin response to oral and i.v. glucose load, due to an increased first phase of insulin secretion (phi 1), a reduced insulin sensitivity (Si) and glucose effectiveness (Sg) in respect to normal-weight subjects. They showed no difference in glucose and insulin area during oral load and in their profiles during i.v. glucose load after naltrexone treatment. Similarly no significant variation in insulin sensitivity and glucose effectiveness was observed. The glucose-induced thermogenesis, measured by indirect calorimetry, was not modified by naltrexone. Therefore our study demonstrates that endogenous opioids do not play any role in the impairment of peripheral insulin sensitivity and energy expenditure in human obesity.

Branched chain amino acids chronic treatment and muscular exercise performance in athletes: a study through plasma acetyl-carnitine levels.

In relation to energy request during physical exercise, muscular tissue Branched Chain Amino Acids (BCAA) are metabolized particularly when the oxidation rises. But in the whole-human body, it is difficult to estimate, in a quantitative sense, the role played by BCAA in sustaining exercise. During a BCAA treatment, made on a group of athletes kept under observation, it was observed, through Conconi's test, that this treatment influenced physical performance. Aim of present work is to investigate if BCAA chronic treatment effect on physiological trial is confirmed on blood circulating biochemical energy parameters and in particular on acetyl-carnitine, since acetyl-linked compounds may be an important biochemical factor.Fourteen athletic well trained male subjects, were randomly divided into two subgroups; a first group was submitted to a chronic treatment (n = 7) of BCAA (oral intake was 0.2 g/Kg die) and a second group, as controls (n = 7), assumed oral placebo. Conconi's test demonstrated a significant difference (p < 0.005) in the exercise performance of the two sub-groups, comparing the measurements of ratios of deflection velocity (V d ), before and after the treatment. Therefore we studied the athletes performing a muscular exercise test (40 Km/h, cycle race, for 90 min) after one month of treatment. During this treatment period the subjects followed a well standardized diet. Samples of blood were drawn before, at the end and during the recovery (60 min) to study if traditional biochemical parameters varied and confirmed the observed differences in Conconi's test. The measurements of concentrations of FFA, KB, free carnitine, acetyl-carnitine and BCAA were performed. Plasma BCAA levels did not demonstrate variations either before or after the exercise performance, or between the two groups. The biochemical factors, substrates and hormones, KB, FFA, lactate, insulin and growth hormone plasma levels did not demonstrate significant differences from the patterns present in literature. Plasma free and acetyl-carnitine followed the well known variations, but only acetyl-carnitine levels demonstrated, at the end increase in acetyl-carnitine levels could be related to a minor fatigue situation and to a larger energy supply availability perhaps present in BCAA treated athletes (Sahlin et al., 1990; May et al., 1989). Both mentioned hypothesis seem in concordance with a smaller acetyl-CoA substrate accumulation, or better for present study, is even more successful with athletes who give a better physical performance. In fact Conconi's test in the two sub-groups of athletes seems to suggest that BCAA treated athletes were able to give a better performance, furthermore out of curiosity we point out that the athletes treated with BCAA won more races than the untreated.We would also like to add in conclusion that although confirming the difficulties of studies in the whole-body, our work gives an interesting clue about the possibility to use acetyl-carnitine plasma levels to understand the biochemical importance of the BCAA as substrate able to influence physical performance, but further research is needed.The phenomenon presence might be showed better perhaps by studying untrained groups during prolonged exercise and with physical performance at exhaustion. If treatment were able to help the physical performance and to shift the fatigue, then confirmation might be a less raised plasma acetyl-carnitine level. In effect blood ammonium levels in present study did not demonstrate any variation in and between sub-groups; this latter observation could be caused by the quantity of work load, and training state of the athletes (Ji et al., 1987; Kirkendall, 1990). Moreover, as observed by Hageloch et al. (1990), the ammonia increases less during prolonged endurance exercise, and in fact the athletes of present study were all middle distance racing cyclists, and the physical performance was a prolonged endurance exercise.

Increased muscular phospholipase A2 activity in diabetic rats.

Anomalies in prostaglandin (PG) synthesis have been suggested in both experimental and human diabetes mellitus; increased levels of plasma and tissue eicosanoids has been recently reported by several investigators. One step in prostaglandin synthesis is the enzymatic hydrolysis of membrane phospholipids by Phospholipase A2 (PLA2). Nevertheless the alternative pathway involving Phospholipase C must be considered. An evaluation of PLA2 activity is therefore a useful method for studying prostaglandin synthesis in the peripheral target tissues of insulin activity. We studied PLA2 activity in normal and diabetic rat muscle. Streptozotocin-induced diabetic rats showed significantly higher muscular PLA2 activity when compared with controls (3.04 x 10(-2) +/- 0.50 x 10(-2) versus 1.34 x 10(-2) +/- 0.35 x 10(-2) arachidonic acid pMol.mg protein-1.min-1 (p less than 0.01). This effect was not observed in diabetic animals successfully treated with insulin (1.78 x 10(-2) +/- 0.5 x 10(-2) versus 1.34 x 10(-2) +/- 0.35 x 10(-2) arachidonic acid pMol.mg protein-1.min-1), and a significant correlation was found between blood glucose and muscular PLA2 activity (r = 0.42; p less than 0.05). Our results clearly show that in streptozotocin-induced diabetic rats muscular PLA2 activity is significantly higher. The relationship between blood glucose levels and muscular PLA2 activity and the decrease of PLA2 activity after insulin treatment suggest that these changes may be related to a defect in insulin effect.

Plasma acetyl-carnitine concentrations during and after a muscular exercise test in patients with liver disease.

In many human tissues, fuel is stored for immediate use, as well as for energy exchange between different parts of the body. Fat and glycogen represent, together with proteins, the principal energy storage materials. During energy requirement, e.g. muscular exercise, glycogen as a local reserve, is used first to supply energy needs. Acetyl-carnitine, as an active molecular group, represents an intermediate substrate, usable directly in the working tissue. The present study investigates whether plasma acetyl-carnitine could be a useful biochemical measure for information on fuel exchange in the body, and whether it is a rapidly available energy source exchangeable among tissues with different metabolic functions, such as muscle and liver. The present study investigated control and hepatopathic subjects after maximal and submaximal muscular exercise. Hepatopathic patients may be a useful model, as liver carnitine metabolism is likely to be impaired. Plasma acetyl-carnitine before, during and after maximal exercise in hepatopathic subjects did not differ, while in normal subjects it increased. After submaximal exercise, acetyl-carnitine increased in patients, as well in controls. In the patients (n = 9) with liver metabolism disorders we observed that during maximal exercise plasma acetyl-carnitine varied from 3.26 +/- 2.18 mumol/l (time 0 min) to 4.30 +/- 2.02 mumol/l (time 20 min) and from 1.99 +/- 1.36 mumol/l to 4.83 +/- 2.60 mumol/l (p less than 0.05) in the controls (n = 7).(ABSTRACT TRUNCATED AT 250 WORDS)

Endocrine pancreatic function in pheochromocytoma.

In six patients with pheochromocytoma oral glucose tolerance test (OGTT) and arginine test were carried out. Blood insulin and glucagon response were investigated. In subjects with adrenal tumor glycemic curve pattern was typical: a rapid and exaggerated increase of glycemia followed by an abrupt drop. Absolute insulinemic response to oral glucose was normal, but inappropriate to glycemic stimulus. Arginine infusion provoked a slightly above normal increase in blood glucose and a normal increase in blood glucagon. In three of the patients studied postoperatively, reduced glycemic response to glucose was observed, whereas there were no evident variations in blood insulin and glucagon response. These data suggest that in pheochromocytoma impaired glucose tolerance is partly due to the reduced insulin response to oral glucose load.