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Aging is characterized by a progressive decline of skeletal muscle (SM) mass and strength which may lead to sarcopenia in older persons. To date, a limited number of studies have been performed in the old SM looking at the whole, complex network of the extracellular matrix (i.e., matrisome) and its aging-associated changes. In this study, skeletal muscle proteins were isolated from whole gastrocnemius muscles of adult (12 mo.) and old (24 mo.) mice using three sequential extractions, each one analyzed by liquid chromatography with tandem mass spectrometry. Muscle sections were investigated using fluorescence- and transmission electron microscopy. This study provided the first characterization of the matrisome in the old SM demonstrating several statistically significantly increased matrisome proteins in the old vs. adult SM. Several proteomic findings were confirmed and expanded by morphological data. The current findings shed new light on the mutually cooperative interplay between cells and the extracellular environment in the aging SM. These data open the door for a better understanding of the mechanisms modulating myocellular behavior in aging (e.g., by altering mechano-sensing stimuli as well as signaling pathways) and their contribution to age-dependent muscle dysfunction.

Hyaluronic acid (HA) is an unbranched polysaccharide particularly abundant in the extracellular matrix (ECM) of soft connective tissues. In humans, about 50% of the total HA in the organism is localized in the skin. HA plays an essential role in the hydration of the ECM, in the regulation of tissue homeostasis, in the resistance to mechanical stimuli/forces, and in the modulation of tissue regeneration. For these reasons, HA is widely used in regenerative medicine and cosmetics. In this study we used an innovative fluid dynamic system to investigate the effects of a cross-linked macrostructural HA formulation on dermal collagen of healthy human skin explants. The good preservation of skin explants provided by the bioreactor allowed applying refined high-resolution microscopy techniques to analyze in situ the HA-induced modifications on the ECM collagen fibrils up to 48 h from the application on the skin surface. Results demonstrated that this HA formulation, commercially proposed for subcutaneous injection, may act on dermal ECM also when applied transcutaneously, improving ECM hydration and modifying the organization of the collagen fibrils. These findings, obtained by the original combination of explanted human skin use with an advanced culture system and multiscale imaging techniques, are consistent with the volumizing and anti-aging effect of HA.

Cerenkov luminescence imaging and Cerenkov photodynamic therapy have been developed in recent years to exploit the Cerenkov radiation (CR) generated by radioisotopes, frequently used in Nuclear Medicine, to diagnose and fight cancer lesions. For in vivo detection, the endpoint energy of the radioisotope and, thus, the total number of the emitted Cerenkov photons, represents a very important variable and explains why, for example, 68Ga is better than 18F. However, it was also found that the scintillation process is an important mechanism for light production. Nanotechnology represents the most important field, providing nanosctructures which are able to shift the UV-blue emission into a more suitable wavelength, with reduced absorption, which is useful especially for in vivo imaging and therapy applications. Nanoparticles can be made, loaded or linked to fluorescent dyes to modify the optical properties of CR radiation. They also represent a useful platform for therapeutic agents, such as photosensitizer drugs for the production of reactive oxygen species (ROS). Generally, NPs can be spaced by CR sources; however, for in vivo imaging applications, NPs bound to or incorporating radioisotopes are the most interesting nanocomplexes thanks to their high degree of mutual colocalization and the reduced problem of false uptake detection. Moreover, the distance between the NPs and CR source is crucial for energy conversion. Here, we review the principal NPs proposed in the literature, discussing their properties and the main results obtained by the proponent experimental groups.

Ozone (O3) is an oxidizing natural gas widely applied as adjunctive therapeutic treatment for a variety of pathological conditions. Currently, O3-based therapies rely on the low-dose concept i.e., the administration of low O3 concentrations able to induce a mild oxidative stress stimulating antioxidant and anti-inflammatory response without causing cell damage. In addition, low O3 concentrations are thought to activate cellular and molecular mechanisms responsible for analgesic and regenerative effects. Due to these properties, in the last decade interest has arisen in the fields of orthopedics and regenerative medicine on the potential of O3 to counteract joint diseases involving cartilage degeneration. In this pilot study, we have explored the anti-degenerative potential of O3 on knee articular cartilage explanted from a healthy adult rabbit and maintained in vitro. Light and transmission electron microscopy were used to monitor chondrocyte and extracellular matrix features of cartilage samples undergoing O3 treatment every three days for two weeks. Results demonstrated that low O3 concentrations act on chondrocytes and the molecular components of the extracellular matrix of articular cartilage explants, significantly improving their preservation under in vitro conditions, likely by promoting both protective and pro-regenerative pathways. This opens promising perspectives for further investigations on the therapeutic potential of O3 for the treatment of cartilage degeneration not only as painkilling and anti-inflammatory agent but also as a cartilage regenerative agent.

Nanoparticles (NPs) include a wide group of small structures composed by very different materials and characterized by peculiar properties that make them suitable for many applications, especially imaging and drug delivery. In this overview we focus on the optical properties of fluorescent NPs available for in vivo, in vitro and ex vivo preclinical studies and detectable with the optical imaging technique alone or in combination with microscopic confocal imaging. We summarize here the basic principles of the optical detection of fluorescent NPs, elucidate which are the current issues to be resolved and possible solutions to achieve the highest sensitivity and specificity for an unbiased analysis. So far NPs application in clinic is in evaluation due to safety questions still unaddressed but in the future they could dramatically improve both preclinical research and patient clinical care. 

Bacterial wilt (BW) caused by is responsible for substantial losses in cultivated potato ( ) crops worldwide. Resistance genes have been identified in wild species; however, introduction of these through classical breeding has achieved only partial resistance, which has been linked to poor agronomic performance. The (At) pattern recognition receptor elongation factor-Tu (EF-Tu) receptor (EFR) recognizes the bacterial pathogen-associated molecular pattern EF-Tu (and its derived peptide elf18) to confer anti-bacterial immunity. Previous work has shown that transfer of AtEFR into tomato confers increased resistance to . Here, we evaluated whether the transgenic expression of would similarly increase BW resistance in a commercial potato line (INIA Iporá), as well as in a breeding potato line (09509.6) in which quantitative resistance has been introgressed from the wild potato relative Resistance to was evaluated by damaged root inoculation under controlled conditions. Both INIA Iporá and 09509.6 potato lines expressing showed greater resistance to , with no detectable bacteria in tubers evaluated by multiplex-PCR and plate counting. Notably, AtEFR expression and the introgression of quantitative resistance from had a significant additive effect in 09509.6-AtEFR lines. These results show that the combination of heterologous expression of with quantitative resistance introgressed from wild relatives is a promising strategy to develop BW resistance in potato.

The expression of leptin and leptin receptor (Ob-R) has been partially elucidated in colon of patients with inflammatory bowel diseases (IBDs), even though leptin is involved in angiogenesis and inflammation. We previously reported overexpression of GLUT5 fructose transporter, in aberrant clusters of lymphatic vessels in lamina propria of IBD and controls. Here, we examine leptin and Ob-R expression in the same biopsies. Specimens were obtained from patients with ulcerative colitis (UC), Crohn’s disease (CD) and controls who underwent screening for colorectal cancer, follow-up after polypectomy or with a history of lower gastrointestinal symptoms. Immunohistochemistry revealed leptin in apical and basolateral membranes of short epithelial portions, Ob-R on the apical pole of epithelial cells. Leptin and Ob-R were also identified in structures and cells scattered in the lamina propria. In UC, a significant correlation between leptin and Ob-R in the lamina propria was found in all inflamed samples, beyond non-inflamed samples of the proximal tract, while in CD, it was found in inflamed distal samples. Most of the leptin and Ob-R positive areas in the lamina propria were also GLUT5 immunoreactive in inflamed and non-inflamed mucosa. A significant correlation of leptin or Ob-R expression with GLUT5 was observed in the inflamed distal samples from UC. Our findings suggest that there are different sites of leptin and Ob-R expression in large intestine and those in lamina propria do not reflect the status of mucosal inflammation. The co-localization of leptin and/or Ob-R with GLUT5 may indicate concomitance effects in colorectal lamina propria areas.

The ability of whole body vibration (WBV) to increase energy expenditure (EE) has been investigated to some extent in the past using short-term single exercises or sets of single exercises. However, the current practice in WBV training for fitness is based on the execution of multiple exercises during a WBV training session for a period of at least 20 min; nevertheless, very limited and inconsistent data are available on EE during long term WBV training session. This crossover study was designed to demonstrate, in an adequately powered sample of participants, the ability of WBV to increase the metabolic cost of exercise vs. no vibration over the time span of a typical WBV session for fitness (20 min). Twenty-two physically active young males exercised on a vibration platform (three identical sets of six different exercises) using an accelerometer-verified vibration stimulus in both the WBV and no vibration condition. Oxygen consumption was measured with indirect calorimetry and expressed as area under the curve (O2(AUC)). Results showed that, in the overall 20-min training session, WBV increased both the O2(AUC) and the estimated EE vs. no vibration by about 22% and 20%, respectively (P<0.001 for both, partial eta squared [η2] ≥0.35) as well as the metabolic equivalent of task (+5.5%, P = 0.043; η2 = 0.02) and the rate of perceived exertion (+13%, P<0.001; ŋ2 = 0.16). Results demonstrated that vibration is able to significantly increase the metabolic cost of exercise in a 20-min WBV training session.

While olfactory deficit is already known to be associated with early-stage Parkinson’s disease (PD), taste perception has not fully clarified so far. In this study, we investigated the taste performance in 61 patients with PD and 66 healthy controls (HC) using the Whole Mouth (WMT) and Taste Strip Tests (TST). In addition, we evaluated their olfactory function by means of the Sniffin’ Sticks Test (SST). TST score was significantly lower in PD patients than in HC (TST score 11.0 ± 2.8 vs. 12.2 ± 2.1; p  < 0.018) while WMT showed no difference. The olfactory evaluation confirmed the results reported in the literature with a significant reduction of the SST score in PD patients than in HC (SST score 7.0 ± 2.8 vs. 11.3 ± 2.8; p  < 0.0001). The conflicting results revealed by TST and WMT could rely on a taste impairment not detectable at supra-threshold concentration of tastes, typical of the daily life. Possible biological correlates of taste impairment in PD are discussed.

The investigation of the absorption of drug delivery systems, designed for the transport of therapeutic molecules inside the body, could be relatively simplified by the fluorophore association and tracking by means of bio-imaging techniques (i.e., optical in vivo imaging or confocal and multiphoton microscopy). However, when a fluorescence signal comes out from the skin, its specific detection can be problematic. Skin high autofluorescence can hinder the observation of administered exogenous fluorophores conjugated to drug delivery systems, making it more challenging to detect their biodistribution. In the present study, we have developed a method based on the spectrofluorometric analysis of skin samples to discriminate the fluorescent signal coming from administered fluorescent molecules from the background. Moreover, we gave a semi-quantitative evaluation of the signal intensity. Thus, we distinguished two gel formulations loading the fluorophore rhodamine B (called GEL RHO and GEL SLN-RHO). The two formulations of gels, one of which containing solid lipid nanoparticles (GEL RHO-SLN), were administered on skin explants incubated in a bioreactor, and the penetration was evaluated at different time points (2 and 6 hours). Cryostatic sections of skin samples were observed with confocal laser scanning microscopy, and a spectrofluorometric analysis was performed. Significantly higher signal intensity in the samples administered with SLN-RHO GEL, with a preferential accumulation in the hair bulbs, was found. Reaching also the deeper layers of the hair shaft after 6 hours, the solid lipid nanoparticles thickened with polymer represent a suitable drug delivery system for transcutaneous administration.