M. De Rossi

Lifelong Physical Exercise Delays Age-Associated Skeletal Muscle Decline

Aging is usually accompanied by a significant reduction in muscle mass and force. To determine the relative contribution of inactivity and aging per se to this decay, we compared muscle function and structure in (a) male participants belonging to a group of well-trained seniors (average of 70 years) who exercised regularly in their previous 30 years and (b) age-matched healthy sedentary seniors with (c) active young men (average of 27 years). The results collected show that relative to their sedentary cohorts, muscle from senior sportsmen have: (a) greater maximal isometric force and function, (b) better preserved fiber morphology and ultrastructure of intracellular organelles involved in Ca(2+) handling and ATP production, (c) preserved muscle fibers size resulting from fiber rescue by reinnervation, and (d) lowered expression of genes related to autophagy and reactive oxygen species detoxification. All together, our results indicate that: (a) skeletal muscle of senior sportsmen is actually more similar to that of adults than to that of age-matched sedentaries and (b) signaling pathways controlling muscle mass and metabolism are differently modulated in senior sportsmen to guarantee maintenance of skeletal muscle structure, function, bioenergetic characteristics, and phenotype. Thus, regular physical activity is a good strategy to attenuate age-related general decay of muscle structure and function (ClinicalTrials.gov: NCT01679977).

D-lysine reduces the non-enzymatic glycation of proteins in experimental diabetes mellitus in rats.

Summaryd-Lysine, the non-physiological isomer of l-lysine, can competitively reduce protein non-enzymatic glycation in vitro. To study the effect of d-lysine in vivo, 6–8-week old Sprague-Dawley rats with streptozotocin-induced diabetes mellitus were treated from diagnosis for 45 days with two daily subcutaneous injections of d-lysine (0.5 g·ml−1·day−1). Another group of diabetic rats was only injected with equal volumes of physiological saline (0.9% NaCl). Glycated haemoglobin was measured by ion exchange chromatography, and glycated serum and lens proteins by boronate affinity gel chromatography. Serum and urinary creatinine concentrations were evaluated by the alkaline-picrate reaction. Urinary lysine concentrations at mid- and end-study were evaluated by cation exchange chromatography. Blood glucose concentrations, serum creatinine levels and creatinine clearances, measured at the end of the study, were similar in both diabetic groups (> 22.0 mmol/l, ≤ 106 μmol/l and ≈ 0.02 ml/s, respectively). Urinary lysine concentration in d-lysine-treated diabetic animals was more than 50-fold higher than in placebo-treated diabetic rats. In d-lysine-treated vs placebo-treated diabetic animals, a statistically significant reduction was found in the levels of glycated haemoglobin (stable HbA1; mean ± SD=3.00±0.74% vs 4.02±0.46%, p<0.05; labile HbA1=3.92±0.89% vs 5.84±0.61%, p<0.005), glycated serum proteins (1.40±0.47% vs 2.52±1.15%, p<0.05) and glycated lens proteins (4.90±0.96% vs 5.98±0.65 %,p<0.05). Thus, d-lysine (i) is not nephrotoxic and (ii) causes a significant reduction of the early glycation products at the protein level. Therefore, the d-amino acid could be useful in attempting to control damaging phenomena associated with or due to an enhanced protein non-enzymatic glycation.

Age-related changes in vasoactive intestinal polypeptide levels and distribution in the rat lung.

Vasoactive intestinal polypeptide (VIP) levels and distribution were studied in the lung of young-adult (3-month-old) and aged (28-month-old) male Wistar rats by radioimmunoassay and immunofluorescence. VIP concentrations were reduced approximately by 60% as the animal ages. The density of VIP-immunoreactive nerve fibres was remarkably reduced within bronchial smooth muscle and bronchial glands. Moreover, the number of VIP-immunoreactive nerve cell bodies located in intraparenchymal ganglia was decreased in old rats. The density of VIP-containing perivascular plexuses was slightly reduced in senescence. The present data are indicative that VIP neuronal system is impaired in the lung of old rats. In view of the significant age-dependent loss of VIP-immunoreactive nerve fibres that supply the bronchial tree and bronchial glands it cannot be excluded that the relaxant action exerted by peptide on airway smooth muscle and the control of bronchial secretion exerted by VIP are impaired in old age.

Anionic versus cationic immunoglobulin clearance in normal subjects : a novel approach to the evaluation of charge permselectivity

The excretion of proteins differing in charge (the different immunoglobulin subclasses) and/or size (albumin, immunoglobulins) were investigated in normal subjects in a number of physiological conditions aiming at the evaluation of renal charge permselectivity. In 101 randomly selected normal subjects the urinary excretion rates of albumin, IgG4 (anionic proteins) and of total IgG (mostly cationic) were evaluated in basal conditions; the protein/creatinine urinary ratio and protein clearances were assessed in part of them. In addition, the intra- and interday variations of protein excretion were evaluated. Protein clearances were measured in a sample group after standardized physical exercise, after an amino acid load, and in orthostatism. Albumin, IgG4 and IgG were assayed using sensitive methods developed in our laboratories. The excretion rate values of albumin, IgG4 and total IgG (median, interquartile range) were 4.36 micrograms/min, (2.58-6.59), 4.25 ng/min (2.6-7.6), and 1.47 micrograms/min (0.85-2.44), respectively. The clearances of the three proteins (mean +/- SD) were 0.13 +/- 0.07, 0.017 +/- 0.012 and 0.14 +/- 0.08 ml/min x 10(-3), respectively. The IgG4/IgG ratio averaged 0.1 and was always below 0.25. Protein excretion rates showed a noticeable variation during the day and from day to day. Physical exercise, the change of posture and the amino acid load significantly increased proteinuria but did not significantly modify the anionic/cationic immunoglobulin ratio. Thus, the anionic/cationic immunoglobulin ratio of about one tenth, substantially stable during dynamic tests, in normal subjects may be considered an index of physiological renal protein charge permselectivity.

A charge selectivity impairment in protein permselectivity is present in type 2 diabetes

A possible loss in kidney charge permselectivity of proteins before any manifestation of nephropathy has been sought in type 2 (non-insulin-dependent) diabetes by assessing the clearances of proteins differing in charge and/or size (anionic and cationic immunoglobulins, albumin). Eighty-five consecutive outpatients with type 2 diabetes were studied and compared with 101 normal subjects. Of the patients, 14.1% wree microalbuminuric and 2.3% macroalbuminuric. A significant increase in protein clearances was observed in diabetic patients in comparison with normal subjects: the median of albumin clearance was 0.09 ml/min, interquartile range (IR) 0.04–0.31 (P<0.01 vs normals); that of anionic immunoglobulins (IgG4) 0.02 ml/min, IR 0.01–0.05 (P<0.005 vs normals); and that of neutral/cationic immunoglobulins (IgG) 0.13 ml/min, IR 0.07–0.19 (P<0.01 vs normals). The anionic/cationic immunoglobulin ratio median was 0.22, IR 0.11–0.43, and exceeded the upper limit of normal values in 29.4% of all patients. IgG4 clearance was positively correlated with albumin clearance (r=0.72) and with IgG clearance (r=0.98). Nevertheless anionic immunoglobulin clearance was increased in a number of patients (17.3%) with normal IgG excretion and even in patiens (15.1%) with normal albumin clearance. Clearances of IgG4 and IgG, but not that of albumin, were correlated with the duration of diabetes. Thus, an increased anionic/cationic IgG ratio in type 2 diabetes highlights a charge selectivity defect in protein permselectivity; this selective proteinuria may reflect more accurately than does microalbuminuria an early kidney abnormality in this form of diabetes.

Vasoactive intestinal polypeptide levels and distribution in the penis of old rats

Vasoactive intestinal polypeptide (VIP) levels and distribution were studied in the penis of young-adult (3-month-old) and old (30-month-old) Wistar rats by radioimmunoassay and immunofluorescence. No significant changes in tissue VIP concentrations or distribution occurred in the penis of old rats compared to young-adult rats. The present data indicate that the age-related impairment of male sexual function is not dependent on modifications of the VIP-ergic innervation of penile tissue.

3H-spiroperidol binding sites in the rabbit superior mesenteric artery. A histoautoradiographic study.

The distribution of 3H-spiroperidol binding sites within rabbit superior mesenteric artery was studied in normal as well as 6-hydroxydopamine (6-OHDA) sympathectomized animals using a histoautoradiographic technique. The labelled drug was located in the three layers of the artery (adventitia, media and intima), with the greater density in the media. In the adventitia the radiolabelled drug was located at the level of fibrous and connective cells. In the media 3H-spiroperidol was bound by smooth muscle cells and is found in the cellular membrane of the same cells. 6-OHDA administration causes an increase in the number of 3H-spiroperidol binding sites in the adventitia and as well as a 20-25% increase in the media. The possible existence of a direct dopaminergic innervation of the superior mesenteric artery is discussed.

Acetylcholinesterase containing nerve fibres in guinea pig ovary

The distribution of acetylcholine esterase (AChE) in the ovary of normal as well as of sympathectomized guinea pigs was studied. In normal animals AChE-positive nerve fibres were found organized in a perivascular plexus. Some nerve fibres reach the corpus luteum and the follicular wall. Chemical sympathectomy performed with the neurotoxin 6-hydroxydopamine caused an almost complete disappearance of AChE positive nerve fibres suggesting that most of the AChE activity of ovarian nerves is localized in adrenergic nerve fibres. Enzyme activity was also present in smooth muscle cells of the vascular tree.

Enhanced nonenzymatic glycation of eye lens proteins in experimental diabetes mellitus: An approach for the study of protein alterations as mediators of normal aging phenomena

The levels of advanced nonenzymatic glycation endproducts (ACE) were investigated by spectrofluorimetry in eye lens proteins obtained from rats with experimental diabetes of 3 and 6 months duration and from normal age-matched control rats. Diabetic animals showed higher AGE levels at both times studied. However the older control animals showed protein ACE levels comparable to those of the experimental 3 months diabetic group. These data suggest that a pathological phenomenon such as enhanced nonenzymatic glycation, associated to diabetic hyperglycemia, can be considered as a process leading to an accelerated aging of proteins. Thus experimental diabetes mellitus may be used as a model to investigate physiological protein senescence.

Direct immunohistochemical detection of binding sites for beta-blocker within rat cerebellum

Abstract The direct immunohistochemical detection of the potent beta-blocker (−)-alprenolol was accomplished in rat cerebellum. (−)-Alprenolol binding sites were found in Purkinje cells and in portions of the molecular and granular layers lying close to the Purkinje cells.