Metabolic Changes Associated With Muscle Expression of SOD1G93A

Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disorder, classified into sporadic or familial forms and characterized by motor neurons death, muscle atrophy, weakness, and paralysis. Among the familial cases of ALS, approximately 20% are caused by dominant mutations in the gene coding for superoxide dismutase (SOD1) protein. Of note, mutant SOD1 toxicity is not necessarily limited to the central nervous system. ALS is indeed a multi-systemic and multifactorial disease that affects whole body physiology and induces severe metabolic changes in several tissues, including skeletal muscle. Nevertheless, whether alterations in the plasticity, heterogeneity, and metabolism of muscle fibers are the result of motor neuron degeneration or alternatively occur independently of it remain to be elucidated. To address this issue, we made use of a mouse model (MLC/SOD1G93A) that overexpresses the SOD1 mutant gene selectively in skeletal muscle. We found an alteration in the metabolic properties of skeletal muscle characterized by alteration in fiber type composition and metabolism. Indeed, we observed an alteration of muscle glucose metabolism associated with the induction of Phosphofructokinases and Pyruvate dehydrogenase kinase 4 expression. The upregulation of Pyruvate dehydrogenase kinase 4 led to the inhibition of Pyruvate conversion into Acetyl-CoA. Moreover, we demonstrated that the MLC/SOD1G93A transgene was associated with an increase of lipid catabolism and with the inhibition of fat deposition inside muscle fibers. All together these data demonstrate that muscle expression of the SOD1G93A gene induces metabolic changes, along with a preferential use of lipid energy fuel by muscle fibers. We provided evidences that muscle metabolic alterations occurred before disease symptoms and independently of motor neuron degeneration, indicating that skeletal muscle is likely an important therapeutic target in ALS

Neuromuscular magnetic stimulation counteracts muscle decline in ALS patients: results of a randomized, double-blind, controlled study

The aim of the study was to verify whether neuromuscular magnetic stimulation (NMMS) improves muscle function in spinal-onset amyotrophic lateral sclerosis (ALS) patients. Twenty-two ALS patients were randomized in two groups to receive, daily for two weeks, NMMS in right or left arm (referred to as real-NMMS, rNMMS), and sham NMMS (sNMMS) in the opposite arm. All the patients underwent a median nerve conduction (compound muscle action potential, CMAP) study and a clinical examination that included a handgrip strength test and an evaluation of upper limb muscle strength by means of the Medical Research Council Muscle Scale (MRC). Muscle biopsy was then performed bilaterally on the flexor carpi radialis muscle to monitor morpho-functional parameters and molecular changes. Patients and physicians who performed examinations were blinded to the side of real intervention. The primary outcome was the change in the muscle strength in upper arms. The secondary outcomes were the change from baseline in the CMAP amplitudes, in the nicotinic ACh currents, in the expression levels of a selected panel of genes involved in muscle growth and atrophy, and in histomorphometric parameters of ALS muscle fibers. The Repeated Measures (RM) ANOVA with a Greenhouse-Geisser correction (sphericity not assumed) showed a significant effect [F(3, 63) = 5.907, p < 0.01] of rNMMS on MRC scale at the flexor carpi radialis muscle, thus demonstrating that the rNMMS significantly improves muscle strength in flexor muscles in the forearm. Secondary outcomes showed that the improvement observed in rNMMS-treated muscles was associated to counteracting muscle atrophy, down-modulating the proteolysis, and increasing the efficacy of nicotinic ACh receptors (AChRs). We did not observe any significant difference in pre- and post-stimulation CMAP amplitudes, evoked by median nerve stimulation. This suggests that the improvement in muscle strength observed in the stimulated arm is unlikely related to reinnervation. The real and sham treatments were well tolerated without evident side effects. Although promising, this is a proof of concept study, without an immediate clinical translation, that requires further clinical validation

Metabolic changes associated with muscle expression of SOD1G93A

Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disorder, classified into sporadic or familial forms and characterized by motor neurons death, muscle atrophy, weakness, and paralysis. Among the familial cases of ALS, approximately 20% are caused by dominant mutations in the gene coding for superoxide dismutase (SOD1) protein. Of note, mutant SOD1 toxicity is not necessarily limited to the central nervous system. ALS is indeed a multi-systemic and multifactorial disease that affects whole body physiology and induces severe metabolic changes in several tissues, including skeletal muscle. Nevertheless, whether alterations in the plasticity, heterogeneity, and metabolism of muscle fibers are the result of motor neuron degeneration or alternatively occur independently of it remain to be elucidated. To address this issue, we made use of a mouse model (MLC/SOD1G93A) that overexpresses the SOD1 mutant gene selectively in skeletal muscle. We found an alteration in the metabolic properties of skeletal muscle characterized by alteration in fiber type composition and metabolism. Indeed, we observed an alteration of muscle glucose metabolism associated with the induction of Phosphofructokinases and Pyruvate dehydrogenase kinase 4 expression. The upregulation of Pyruvate dehydrogenase kinase 4 led to the inhibition of Pyruvate conversion into Acetyl-CoA. Moreover, we demonstrated that the MLC/SOD1G93Atransgene was associated with an increase of lipid catabolism and with the inhibition of fat deposition inside muscle fibers. All together these data demonstrate that muscle expression of the SOD1G93Agene induces metabolic changes, along with a preferential use of lipid energy fuel by muscle fibers. We provided evidences that muscle metabolic alterations occurred before disease symptoms and independently of motor neuron degeneration, indicating that skeletal muscle is likely an important therapeutic target in ALS

Taurine Administration Counteracts Aging-Associated Impingement of Skeletal Muscle Regeneration by Reducing Inflammation and Oxidative Stress

Abstract: Sarcopenia, which occurs during aging, is characterized by the gradual loss of skeletal muscle mass and function, resulting in a functional decline in physical abilities. Several factors contribute to the onset of sarcopenia, including reduced regenerative capacity, chronic low-grade inflammation, mitochondrial dysfunction, and increased oxidative stress, leading to the activation of catabolic pathways. Physical activity and adequate protein intake are considered effective strategies able to reduce the incidence and severity of sarcopenia by exerting beneficial effects in improving the muscular anabolic response during aging. Taurine is a non-essential amino acid that is highly expressed in mammalian tissues and, particularly, in skeletal muscle where it is involved in the regulation of biological processes and where it acts as an antioxidant and anti-inflammatory factor. Here, we evaluated whether taurine administration in old mice counteracts the physiopathological effects of aging in skeletal muscle. We showed that, in injured muscle, taurine enhances the regenerative process by downregulating the inflammatory response and preserving muscle fiber integrity. Moreover, taurine attenuates ROS production in aged muscles by maintaining a proper cellular redox balance, acting as an antioxidant molecule. Although further studies are needed to better elucidate the molecular mechanisms responsible for the beneficial effect of taurine on skeletal muscle homeostasis, these data demonstrate that taurine administration ameliorates the microenvironment allowing an efficient regenerative process and attenuation of the catabolic pathways related to the onset of sarcopenia

La reazione polimerasica a catena (PCR) nella diagnosi delle infezioni genitali da Papillomavirus umano (HPV)

In a three year time (2001-2003) we evaluated the use of HPV testing and typing in 520 cytologically abnormal women (322 LSIL, 198 HSIL), in 25 squamous and 18 adeno invasive cancers. HPV testing was performed by polymerase chain reaction (PCR) using L1 consensus primers; HPV typing was performed using specific primers for low and high risk types. The prevalence of high risk HPV was: LSIL 41%; HSIL 74%; squamous invasive cancers 96% and adeno 100%. HPV 16 is the most represented type in all lesions. We also evaluated the usefulness of p53 typing in the early identification of women at risk for cervical cancer

Detection of Gardnerella vaginalis, Candida spp. and Trichomonas vaginalis DNA in symptomatic women

While vaginitis caused by Trichomonas vaginalis is now less frequent, fungal Candida spp. infections are frequently found and the bacterial vaginosis is one of the most common vaginal diseases caused by anaerobic microorganisms such as Gardnerella vaginalis. Purpose of this study is to evaluate the usefulness of a rapid molecular test for the diagnosis of vaginitis/bacterial vaginosis in symptomatic women. In our clinic, between January 2008 - June 2009, we admitted 1592 (388 were pregnant) symptomatic women with a specific request to test them for fungi, Trichomonas and Gardnerella on vaginal fluid. The samples were tested with the kit Affirm (Becton Dickinson) that provides results in 40 minutes and allows the simultaneous identification of the DNA of Gardnerella vaginalis, Candida spp. and Trichomonas vaginalis. One hundred and eight out of 388 pregnant women were positive only for Gardnerella; 53 for Candida and Gardnerella; 59 were positive only for Candida and 10 for Trichomonas. As to the remaining 1204 not pregnant patients, 356 were positive only for Gardnerella; 98 for Candida and Gardnerella, 143 were positive only for Candida and 21 for Trichomonas.A simultaneous positivity for Trichomonas and Candida or for Trichomonas and Gardnerella has not been observed in any case. Molecular testing is obviously more sensitive and specific than culture method and microscopic research, especially for the detection of Gardnerella. It also enables differential diagnosis between bacterial vaginosis and vaginitis and therefore allows a targeted therapeutic intervention

Circulating myomiRs in Muscle Denervation: From Surgical to ALS Pathological Condition

ALS is a fatal neurodegenerative disease that is associated with muscle atrophy, motoneuron degeneration and denervation. Different mechanisms have been proposed to explain the pathogenesis of the disease; in this context, microRNAs have been described as biomarkers and potential pathogenetic factors for ALS. MyomiRs are microRNAs produced by skeletal muscle, and they play an important role in tissue homeostasis; moreover, they can be released in blood circulation in pathological conditions, including ALS. However, the functional role of myomiRs in muscle denervation has not yet been fully clarified. In this study, we analyze the levels of two myomiRs, namely miR-206 and miR-133a, in skeletal muscle and blood samples of denervated mice, and we demonstrate that surgical denervation reduces the expression of both miR-206 and miR-133a, while miR-206 but not miR-133a is upregulated during the re-innervation process. Furthermore, we quantify the levels of miR-206 and miR-133a in serum samples of two ALS mouse models, characterized by different disease velocities, and we demonstrate a different modulation of circulating myomiRs during ALS disease, according to the velocity of disease progression. Moreover, taking into account surgical and pathological denervation, we describe a different response to increasing amounts of circulating miR-206, suggesting a hormetic effect of miR-206 in relation to changes in neuromuscular communication

HPV DNA genotyping test in low and high grade intraephitelial lesions

The human papillomavirus (HPV) is certainly one of the few viruses associated with cancer and is present in a significant number of cancers of the anus, penis, vagina, vulva and in almost all cervical cancers.The aim of our work is to evaluate the impact of genotypes high-risk oncogenic (HR) in low-grade (LSIL) and high grade (HSIL) intraepithelial lesions. In it are also evaluated the impact of genotypes low oncogenic risk (LR) and the percentage of non-detection (NR) Papillomavirus DNA Between January 2008 and June 2009 we submitted for HPV DNA genotyping test 392 patients, seronegative for HIV1/2, of which 278 with LSIL and 114 with HSIL. In 250 patients tested positive for HPV DNA HR the type 16 is present in 102 patients (41%), the 18 in 62 (25%), the 31 in 37 patients (15%), the 33 in 13 patients (5%); also was limited the simultaneous presence of 16 and 18 (13 patients 5%) The search for HPV DNA is a modern method of study of cervical pathology. Optimizes the follow-up of intraepithelial lesions and the therapeutic intervention, avoiding the dangers of over/ undertreatment and allows to monitor women treated for cervical pathology

Polyoma BK virus infection in renal transplant recipients

Several studies reported a correlation between the human Polyomavirus BK (BKV) and interstitial nephritis (PVN: Polyoma Virus Nephropaty) in renal transplant recipients in whom immunosuppressive treatment is thought to facilitate or induce reactivation of the virus. In the present study we monthly evaluated the presence of BKV-DNA in plasma and urine of 29 kidney transplant recipients. We used a nested PCR for BKV-DNA screening in plasma and urine and a quantitative assay Real Time PCR in case of a positive screening result. The viral DNA has been detected in 48% of the patients samples: only in urine of six patients; in plasma of four and in both of two. Immunosuppressive therapy has been decreased in positive patients. The kidney loss occurred just in the two patients with high viral load in plasma and urine. The definitive diagnosis of BKV Nephropaty requires allograft biopsy though biomolecular test for BKV-DNA in urine (viruria) and in plasma (viremia) could be very important non-invasive method for the early diagnosis of PVN

Qualità dell’aria negli ambienti confinati: le attività dell’ISPRA

Questo volume include le relazioni presentate durante il workshop sull’inquinamento indoor, che ha avuto un duplice scopo di: 1) fornire una panoramica delle iniziative a livello nazionale e internazionale in termini di studi, strategie di campionamento degli inquinanti chimici e biologici, e delle norme per prevenire e/o ridurre la contaminazione dell’aria indoor; 2) presentare le esperienze maturate dai componenti del Gruppo di Studio Nazionale sull’Inquinamento Indoor e dalle principali istituzioni nazionali.This document reports the presentations at the workshop on indoor air pollution, which had a dual aim: 1) to provide an overview of initiatives at national and international level, in terms of ongoing studies and strategies for sampling and analysing indoor air chemical and biological contaminants, and technical regulations to prevent and/or to reduce indoor air contamination; 2) to report the experiences in the field by the members of the National Working Group on indoor air and main national institutions as well