Extracellular vesicles from mesenchymal stem cells: towards novel therapeutic strategies for neurodegenerative diseases

Neurodegenerative diseases are fatal disorders of the central nervous system (CNS) which currently lack effective treatments. The application of mesenchymal stem cells (MSCs) represents a new promising approach for treating these incurable disorders. Growing evidence suggest that the therapeutic effects of MSCs are due to the secretion of neurotrophic molecules through extracellular vesicles. The extracellular vesicles produced by MSCs (MSC-EVs) have valuable innate properties deriving from parental cells and could be exploited as cell-free treatments for many neurological diseases. In particular, thanks to their small size, they are able to overcome biological barriers and reach lesion sites inside the CNS. They have a considerable pharmacokinetic and safety profile, avoiding the critical issues related to the fate of cells following transplantation. This review discusses the therapeutic potential of MSC-EVs in the treatment of neurodegenerative diseases, focusing on the strategies to further enhance their beneficial effects such as tracking methods, bioengineering applications, with particular attention to intranasal delivery as a feasible strategy to deliver MSC-EVs directly to the CNS in an effective and minimally invasive way. Current progresses and limiting issues to the extent of the use of MSC-EVs treatment for human neurodegenerative diseases will be also revised

Nucleophosmin and its AML-associated mutant regulate c-Myc turnover through Fbw7γ

Mutations leading to aberrant cytoplasmic localization of nucleophosmin (NPM) are the most frequent genetic alteration in acute myelogenous leukemia (AML). NPM binds the Arf tumor suppressor and protects it from degradation. The AML-associated NPM mutant (NPMmut) also binds p19Arf but is unable to protect it from degradation, which suggests that inactivation of p19Arf contributes to leukemogenesis in AMLs. We report here that NPM regulates turnover of the c-Myc oncoprotein by acting on the F-box protein Fbw7γ, a component of the E3 ligase complex involved in the ubiquitination and proteasome degradation of c-Myc. NPM was required for nucleolar localization and stabilization of Fbw7γ. As a consequence, c-Myc was stabilized in cells lacking NPM. Expression of NPMmut also led to c-Myc stabilization because of its ability to interact with Fbw7γ and delocalize it to the cytoplasm, where it is degraded. Because Fbw7 induces degradation of other growth-promoting proteins, the NPM–Fbw7 interaction emerges as a central tumor suppressor mechanism in human cancer

Elongated styloid process evaluation on digital panoramic radiograph in a North Italian population

The aim of this study is to evaluate the prevalence of elongated styloid process in digital panoramic radiographs in a North Italian population in relation to age, gender and side. This study was performed as a retrospective analysis on digital panoramic radiographs of 600 (271 males and 329 females) Italian patients between 6 and 87 years old. The styloid process length were measured using the measuring tool of Sidexis Software. It was measured from the point where it left the temporal bone plate to its tip. Styloid processes measuring more than 30 mm were considered elongated. Chi-squared and Fisher tests were used and the test is considered significant if the p-value is lower or equal to 0.05. Thirty-three per cent of the patients showed an elongated styloid process. Seventeen per cent were elongated on both right and left side, fifteen point nine per cent were elongated only in one side. The prevalence of elongated styloid process was high and a progressive increase of the elongation prevalence was found in older groups

The use of the laser confocal scanning microscopy to measure resin remnants on customized lingual bracket

Background: The study aimed to evaluate the permanence of resin and enamel remains on lingual brackets at the end of orthodontic treatment and after the debonding procedure. The evaluation of resin remnants on customized lingual brackets bases has never done before in other studies because they are curved, and traditional techniques are not applicable. Methods: The sample consisted of 100 lingual brackets (25 incisors, 25 canines, 25 premolars, 25 molars) scanned with a confocal laser microscope (OLS4000). We measured the brackets' surface and the area of resin remnants with the software of the microscope. Median and quartiles were presented to describe the data. ARI calculation was indirect for each tooth, measuring the resin remnants to the total surface of the bracket. The Kruskal-Wallis test and Fisher test were applied respectively to compare the percentages of remnants and the frequencies of the ARI between the four groups. Results: After the analyses, 13 brackets had no adhesive remnants (ARI 0), 29 brackets had less than 50% of resin remnants (ARI 1), 50 brackets had more than 50% of resin remnants (ARI 2), and 8 brackets had 100% of adhesive (ARI 3). Canines brackets presented the lower amount of resin followed by premolars, incisors, and molars. Conclusion: Lingual brackets showed a high frequency of ARI = 2. The median percentage of the bracket surface covered by resin was 41%. We observed a slight tendency of more resin remnants on molar brackets, due to half-pad configuration. The authors suggest paying attention during the debonding procedure of molar brackets since a stronger connection between the adhesive and the bracket mesh means a higher risk of enamel damage

Navicular tenosuspension with anterior tibialis tendon (Young procedure) associated to calcaneo-stop for the treatment of paediatric flexible flatfoot: clinical and ultrasound study

Background and aim of the work: Flexible flatfoot is one of the most common deformities in pediatric orthopaedics. Arthroeresis procedures are designed to correct this deformity. Among them, calcaneostop is a procedure with both biomechanical and proprioceptive properties. There could be other surgical procedure combined, such as a percutaneous Achilles tendon lengthening and the Gould tibialis posterior retension or Young tibialis anterior navicular tenosuspension. This study analyzed the clinical and sonographic results of 36 patients following flexible flatfoot surgical treatment with a calcaneo-stop arthroeresis combined with Achilles lengthening and a Young procedure. Methods: From March 2001 to August 2014, 36 patients (54 feet) were treated with calcaneo-stop arthroeresis, percutaneous Achilles tendon lengthening and Young\u2019s tenosuspension. The clinical assessment and a sonography of the anterior tibialis tendon (ATT) were performed in all patients. Results: The average follow-up was 7.4 years (range 8 months-14 years) with a satisfactory outcome in 51 feet (94.5%). No major and minor complications were observed. In four cases the calcaneo-stop was removed for pain and low tolerance of the patient. The AOFAS score and the talocalcaneal angle did not have statistically significant in case of ATT was or not still inserted in the navicular at the follow-up. Conclusions: The calcaneo-stop procedure is a simple, reliable and minimally invasive procedure for the treatment of pediatric flexible flatfoot. Although the indications for the Young tenosuspension as an isolated procedure is very narrow, it can still be an effective procedure when combined to calcaneo-stop. The key to appropriate utilization is a thorough understanding of the biomechanics of the foot function and a specific appreciation of the function of the ATT

Pain Modulation after Oromucosal Cannabinoid Spray (SATIVEX®) in Patients with Multiple Sclerosis: A Study with Quantitative Sensory Testing and Laser-Evoked Potentials

Background. Delta-9-tetrahydrocannabinol (THC)/cannabidiol (CBD) (nabiximols or Sativex®) is an oromucosal spray formulation containing THC and CBD at an approximately 1:1 fixed ratio. Its administration for the treatment of pain in patients with multiple sclerosis (MS) has been established. MS patients generally complain of different kinds of pain, including spasticity-related and neuropathic pain. In this study, we compared and evaluated pain modulation and thermal/pain threshold of MS patients before and after THC/CBD administration. Methods. 19 MS patients underwent clinical examination, numerical rating scale (NRS), quantitative sensory testing (QST), and laser-evoked potentials (LEPs) before and after 1 month of therapy. Psychophysiological and neurophysiological data were compared to sex- and age-matched controls. Results. Patients reported a significant reduction in pain. We found statistically significant differences in LEP parameters between patients and controls but no significant change in LEP measures after THC/CBD therapy. Cold and heat detection thresholds were altered in patients but did not change after THC/CBD therapy. There was a significant increase in cold pain threshold by hand stimulation and a significant reduction in abnormal cold perception thresholds. Conclusions. Our results indicate that Sativex® therapy provides pain relief in MS patients and suggest that it might modulate peripheral cold-sensitive TRP channels

PRECIPITAÇÃO MENSAL E ANUAL PROVÁVEL NO ESTADO DE SANTA CATARINA

A chuva é um dos fatores meteorológicos que apresenta grande variação espacial e temporal. Diversos trabalhos ao ar livre são afetados de forma direta e indireta pela sua ausência ou excesso de precipitação. Este trabalho tem como objetivo caracterizar a distribuição sazonal e espacial da precipitação mensal e anual no Estado de Santa Catarina. Foram utilizados os dados de precipitação diária do período de 1970 a 2012 de 92 estações pluviométricas distribuídas no estado de Santa Catarina. Para estimar a precipitação provável em período mensal foi usada a distribuição gama e para a precipitação anual foram avaliadas as distribuições normal e gama. A aderência das séries de precipitação às distribuições de probabilidade foi avaliada pelo teste de Kolmogorov-Smirnov, ao nível de significância de 5%. Para representar a variação espacial e sazonal da precipitação foram gerados mapas das variáveis analisadas usando o software ArcGIS. Foram determinados os mapas de precipitação e anual  com probabilidades de 25%, 50% e 75 %, em que se pode observar a variação sazonal e também espacial da precipitação no estado de Santa Catarina.  Com base nos dados, observou-se que a distribuição Gama se mostrou adequada para estimar as probabilidades de chuva mensal. Para as séries de precipitação anual, embora ambas distribuições foram consideradas adequadas verificou-se que em 75% das estações o ajuste obtido com a distribuição gama  foi melhor que a distribuição normal. A precipitação mensal e anual apresenta grande variação espacial e sazonal no estado de Santa Catarina. Nos meses de outubro a março as maiores precipitações mensais ocorrem na região do litoral norte e menor no litoral sul. Nos meses de abril a setembro observa-se maiores precipitações na região oeste do estad

ASC-exosomes ameliorate the disease progression in SOD1(G93A) murine model underlining their potential therapeutic use in human ALS

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive degeneration of motoneurons. To date, there is no effective treatment available. Exosomes are extracellular vesicles that play important roles in intercellular communication, recapitulating the effect of origin cells. In this study, we tested the potential neuroprotective effect of exosomes isolated from adipose-derived stem cells (ASC-exosomes) on the in vivo model most widely used to study ALS, the human SOD1 gene with a G93A mutation (SOD1(G93A)) mouse. Moreover, we compared the effect of two different routes of exosomes administration, intravenous and intranasal. The effect of exosomes administration on disease progression was monitored by motor tests and analysis of lumbar motoneurons and glial cells, neuromuscular junction, and muscle. Our results demonstrated that repeated administration of ASC-exosomes improved the motor performance; protected lumbar motoneurons, the neuromuscular junction, and muscle; and decreased the glial cells activation in treated SOD1(G93A) mice. Moreover, exosomes have the ability to home to lesioned ALS regions of the animal brain. These data contribute by providing additional knowledge for the promising use of ASC-exosomes as a therapy in human ALS

Extracellular vesicles from adipose mesenchymal stem cells target inflamed lymph nodes in experimental autoimmune encephalomyelitis

Background aims: Adipose mesenchymal stem cells (ASCs) represent a promising therapeutic approach in inflammatory neurological disorders, including multiple sclerosis (MS). Recent lines of evidence indicate that most biological activities of ASCs are mediated by the delivery of soluble factors enclosed in extracellular vesicles (EVs). Indeed, we have previously demonstrated that small EVs derived from ASCs (ASC-EVs) ameliorate experimental autoimmune encephalomyelitis (EAE), a murine model of MS. The precise mechanisms and molecular/cellular target of EVs during EAE are still unknown. Methods: To investigate the homing of ASC-EVs, we intravenously injected small EVs loaded with ultra-small superparamagnetic iron oxide nanoparticles (USPIO) at disease onset in EAE-induced C57Bl/6J mice. Histochemical analysis and transmission electron microscopy were carried out 48 h after EV treatment. Moreover, to assess the cellular target of EVs, flow cytometry on cells extracted ex vivo from EAE mouse lymph nodes was performed. Results: Histochemical and ultrastructural analysis showed the presence of labeled EVs in lymph nodes but not in lungs and spinal cord of EAE injected mice. Moreover, we identified the cellular target of EVs in EAE lymph nodes by flow cytometry: ASC-EVs were preferentially located in macrophages, with a consistent amount also noted in dendritic cells and CD4+ T lymphocytes. Conclusions: This represents the first direct evidence of the privileged localization of ASC-EVs in draining lymph nodes of EAE after systemic injection. These data provide prominent information on the distribution, uptake and retention of ASC-EVs, which may help in the development of EV-based therapy in MS