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Ghrelin, an acylated peptide produced predominantly by the stomach, has been discovered to be a natural ligand of the growth hormone secretagogue receptor 1a (GHS-R1a). It is localized in distinct cells of the gastric mucosa, mainly distributed in the mid portion of the oxyntic gland characterized by P/D1 granules in man and X/A-like granules in rodents. The ghrelin cell represents the second most frequent endocrine cell type after the enterochromaffin-like cells in gastric oxyntic mucosa, pointing to a potentially relevant role in the physiology of the stomach. Ghrelin has no relevant homology with any known gastrointestinal peptide and displays strong GH-releasing activity both in animals and in humans. However, in addition to stimulating GH secretion, ghrelin possesses several other endocrine and extraendocrine biological activities that are explained by the widespread distribution of ghrelin and GHSR1a expression. In the rat, ghrelin exerts a control in gastric acid secretion and motility: the gastric acid secretion is stimulated by peripheral administration of high doses of ghrelin, but inhibited by very low doses of ghrelin delivered into the central nervous system. Moreover, ghrelin provides a potent and dose-related gastroprotective action against ethanol- and stress-induced gastric ulcers. The integrity of both nitric oxide (NO) system and capsaicin afferent nerves are required for the gastroprotective effect of ghrelin, whereas the vagus nerve might be involved in conveying ghrelinergic signal from periphery to the brain. In addition, prostaglandins derived by the constitutive cyclooxygenase (COX) activity are essential for the protective activity of ghrelin in ethanol and stress-induced gastric lesions. Given its prevailing role in physiological and pathophysiological gastric function, the discovery of ghrelin will open new perspectives and potential clinical implications in the gastroenteric field.

The transcription factor RUNX1, a pivotal regulator of HSCs and haematopoiesis, is a frequent target of chromosomal translocations, point mutations or altered gene/protein dosage. These modifications lead or contribute to the development of myelodysplasia, leukaemia or platelet disorders. A better understanding of how regulatory elements contribute to fine‐tune the RUNX1 expression in haematopoietic tissues could improve our knowledge of the mechanisms responsible for normal haematopoiesis and malignancy insurgence. The cohesin RAD21 was reported to be a regulator of RUNX1 expression in the human myeloid HL60 cell line and during primitive haematopoiesis in zebrafish. In our study, we demonstrate that another cohesin, NIPBL, exerts positive regulation of RUNX1 in three different contexts in which RUNX1 displays important functions: in megakaryocytes derived from healthy donors, in bone marrow samples obtained from adult patients with acute myeloid leukaemia and during zebrafish haematopoiesis. In this model, we demonstrate that alterations in the zebrafish orthologue nipblb reduce runx1 expression with consequent defects in its erythroid and myeloid targets such as gata1a and spi1b in an opposite way to rad21. Thus, also in the absence of RUNX1 translocation or mutations, additional factors such as defects in the expression of NIPBL might induce haematological diseases.

Amyotrophic lateral sclerosis (ALS) is an incurable motor neuron disease whose etiology remains unresolved; nonetheless, mutations of superoxide dismutase 1 (SOD1) have been associated with several variants of ALS. Currently available pharmacologic interventions are only symptomatic and palliative in effect; therefore, there is a pressing demand for more effective drugs. This study examined potential therapeutic effects of growth hormone secretagogues (GHSs), a large family of synthetic compounds, as possible candidates for the treatment of ALS. Human neuroblastoma cells expressing the SOD1-G93A mutated protein (SH-SY5Y SOD1G93A cells) were incubated for 24 h with H2O2 (150 µM) in the absence, or presence, of GHS (1 µM), in order to study the protective effect of GHS against increased oxidative stress. The two GHSs examined in this study, hexarelin and JMV2894, protected cells from H2O2-induced cytotoxicity by activating molecules that regulate apoptosis and promote cell survival processes. These findings suggest the possibility of developing new GHS-based anti-oxidant and neuroprotective drugs with improved therapeutic potential. Further investigations are required for the following: (i) to clarify GHS molecular mechanisms of action, and (ii) to envisage the development of new GHSs that may be useful in ALS therapy.

IntroductionGrowth hormone secretagogues (GHSs) exert multiple actions, being able to activate GHS-receptor 1a, control inflammation and metabolism, to enhance GH/insulin-like growth factor-1 (IGF-1)-mediated myogenesis, and to inhibit angiotensin-converting enzyme. These mechanisms are of interest for potentially targeting multiple steps of pathogenic cascade in Duchenne muscular dystrophy (DMD).MethodsHere, we aimed to provide preclinical evidence for potential benefits of GHSs in DMD, via a multidisciplinary in vivo and ex vivo comparison in mdx mice, of two ad hoc synthesized compounds (EP80317 and JMV2894), with a wide but different profile. 4-week-old mdx mice were treated for 8 weeks with EP80317 or JMV2894 (320 µg/kg/d, s.c.).ResultsIn vivo, both GHSs increased mice forelimb force (recovery score, RS towards WT: 20% for EP80317 and 32% for JMV2894 at week 8). In parallel, GHSs also reduced diaphragm (DIA) and gastrocnemius (GC) ultrasound echodensity, a fibrosis-related parameter (RS: ranging between 26% and 75%). Ex vivo, both drugs ameliorated DIA isometric force and calcium-related indices (e.g., RS: 40% for tetanic force). Histological analysis highlighted a relevant reduction of fibrosis in GC and DIA muscles of treated mice, paralleled by a decrease in gene expression of TGF-β1 and Col1a1. Also, decreased levels of pro-inflammatory genes (IL-6, CD68), accompanied by an increment in Sirt-1, PGC-1α and MEF2c expression, were observed in response to treatments, suggesting an overall improvement of myofiber metabolism. No detectable transcript levels of GHS receptor-1a, nor an increase of circulating IGF-1 were found, suggesting the presence of a novel receptor-independent mechanism in skeletal muscle. Preliminary docking studies revealed a potential binding capability of JMV2894 on metalloproteases involved in extracellular matrix remodeling and cytokine production, such as ADAMTS-5 and MMP-9, overactivated in DMD.DiscussionOur results support the interest of GHSs as modulators of pathology progression in mdx mice, disclosing a direct anti-fibrotic action that may prove beneficial to contrast pathological remodeling.

This study investigated modifications of microRNA expression profiles in knee synovial fluid of patients with osteoarthritis (OA) and rupture of the anterior cruciate ligament (ACL). Twelve microRNAs (26a-5p, 27a-3p, let7a-5p, 140-5p, 146-5p, 155-5p, 16-5p,186-5p, 199a-3p, 210-3p, 205-5p, and 30b-5p) were measured by real-time quantitative polymerase chain reaction (RT-qPCR) in synovial fluids obtained from 30 patients with ACL tear and 18 patients with knee OA. These 12 miRNAs were chosen on the basis of their involvement in pathological processes of bone and cartilage. Our results show that miR-26a-5p, miR-186-5p, and miR-30b-5p were expressed in the majority of OA and ACL tear samples, whereas miR-199a-3p, miR-210-3p, and miR-205-5p were detectable only in a few samples. Interestingly, miR-140-5p was expressed in only one sample of thirty in the ACL tear group. miR-140-5p has been proposed to modulate two genes (BGN and COL5A1100) that are involved in ligamentous homeostasis; their altered expression could be linked with ACL rupture susceptibility. The expression of miR-30b-5p was higher in OA and chronic ACL groups compared to acute ACL samples. We provide evidence that specific miRNAs could be detected not only in synovial fluid of patients with OA, but also in post-traumatic ACL tears.

Made available in DSpace on 2022-05-01T10:51:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-01 Evidence regarding the effect of the onlay preparation design for different CAD/CAM restorative materials considering the preservation of cusps is lacking. Molars were 3D-modeled in four preparation designs for onlay restoration: Traditional design with functional cusp coverage (TFC), non-retentive design with functional cusp coverage (NFC), traditional design with non-functional cusp coverage (TNFC) and non-retentive design with non-functional cusp coverage (NNFC). The restorations were simulated with two CAD/CAM restorative materials: LD—lithium disilicate (IPS e.max CAD) and RC—resin composite (GrandioBloc). A 100 N axial load was applied to the occlusal surface, simulating the centric contact point. Von Mises (VM) and maximum principal (Pmax) stress were evaluated for restorations, cement layer and dental substrate. The non-retentive preparation design reduced the stress concentration in the tooth structure in comparison to the conventional retentive design. For LD onlays, the stress distribution on the restoration intaglio surface showed that the preparation design, as well as the prepared cusp, influenced the stress magnitude. The non-retentive preparation design provided better load distribution in both restorative materials and more advantageous for molar structure. The resin composite restoration on thenon-functional cusp is recommended when the functional cusp is preserved in order to associate conservative dentistry and low-stress magnitude. School of Dentistry Universidad Espíritu Santo Institute of Science and Technology São Paulo State University (Unesp) São José dos Campos Department of Dental Materials Academic Centre for Dentistry Amsterdam (ACTA) University of Amsterdam and Vrije Universiteit Amsterdam Department of Dentistry Center for Biological and Health Sciences Western Paraná State University (Unioeste) Fraunhofer JL IDEAS Department of Industrial Engineering University of Naples Federico II School of Dentistry University of Naples Federico II Institute of Science and Technology São Paulo State University (Unesp) São José dos Campos

Palmitoylethanolamide (PEA) is an endogenous lipid produced on demand by neurons and glial cells that displays neuroprotective properties. It is well known that inflammation and neuronal damage are strictly related processes and that microglia play a pivotal role in their regulation. The aim of the present work was to assess whether PEA could exert its neuroprotective and anti-inflammatory effects through the modulation of microglia reactive phenotypes. In N9 microglial cells, the pre-incubation with PEA blunted the increase of M1 pro-inflammatory markers induced by lipopolysaccharide (LPS), concomitantly increasing those M2 anti-inflammatory markers. Images of microglial cells were processed to obtain a set of morphological parameters that highlighted the ability of PEA to inhibit the LPS-induced M1 polarization and suggested that PEA might induce the anti-inflammatory M2a phenotype. Functionally, PEA prevented Ca2+ transients in both N9 cells and primary microglia and antagonized the neuronal hyperexcitability induced by LPS, as revealed by multi-electrode array (MEA) measurements on primary cortical cultures of neurons, microglia, and astrocyte. Finally, the investigation of the molecular pathway indicated that PEA effects are not mediated by toll-like receptor 4 (TLR4), on the contrary, a partial involvement of cannabinoid type 2 receptor (CB2R) was shown by using a selective receptor inverse agonist.

Background: In neurodegenerative diseases, hydrogen peroxide (H2O2) and the production of reactive oxygen species (ROS) have been implicated in the activation of well-known inflammatory signalling pathways. Microglial cells can assume two opposite phenotypes: M2 anti-inflammatory phenotype, in the early stages of neurodegenerative diseases, and the M1 pro-inflammatory subtype that become prevalentwith the diseases progression. Recently, different studies have demonstrated that microglia communicate with neuronal and non neuronal cells by releasing microvescicles (MVs) that contribute to disease progression. Targeting microglia M2 polarization represent an interesting therapeutic approach to inhibit local neurodegeneration, for example inducing the release of insulin growth factor-1 (IGF-1), a beneficial factor against inflammation. Our previous studies have demonstrated that hexarelin, a well-known growth hormone secretagogues (GHS), is endowed with antioxidant, anti-inflammatory and neuroprotective activities, promotes neurogenesis and inhibits cytotoxic effects of β-amyloid in N9 cells, but can also stimulate the release of IGF-1. In this study, we explored the ability of hexarelin to modulated the phenotype of N9 cells and how the induction of M2 microglia phenotype can oppose H2O2-induced neurotoxicity in mouse neuroblastoma Neuro2A cells. Methods: N9 cells were incubated with hexarelin 10-6 M for 24 h. Cells were used to quantify mRNA levels of iNOS and Arg1 for the evaluation of M1/M2 polarization by Real Time PCR, while medium was collected for subsequent experiments. N9 collected media were processed by two centrifugations at 1000xg for 10 and 20 minutes, respectively. The supernatant was ultracentrifugated at 110’000xg for 70 minutes and the pellet resuspended. Before use, MVs were quantified by NanoSight instrument. Neuro2A cells were treated with H2O2 100 µM for 24 h. At the end of the treatment, H2O2 was removed and substituted with N9 conditioned media, purified MVs, or N9 conditioned media deprived of MVs for 24 h. mRNA levels of Bax, BCL-2, caspase-3 and caspase-7 were measured by Real Time PCR. Results: Hexarelin-treated N9 cells showed a prevalence of M2 anti-inflammatory phenotype, with the down-regulation of iNOS and the up-regulation of Arg1 mRNA levels. H2O2 100 µM induced the activation of apoptosis pathway: the mRNA level of pro-apoptotic Bax significantly increased, as well as the levels of caspase-3 and caspase-7, while BCL-2 mRNA levels were reduced. N9 collected media and MVs extracted inhibited the apoptosis activation: Bax, caspase-3 and caspase-7 mRNA levels were reduced while anti-apoptotic BCL-2 mRNA levels were increased. Conclusion: Our results suggest that hexarelin modulated the phenotype of N9 microglial cells, inducing a polarization toward the M2 anti-inflammatory phenotype. Medium derived by hexarelin-treated N9 cells and MVs reduced the toxic effect of H2O2. Further experiments are needed to clarify the composition of MVs underlying the neuroprotective effects.

Hexarelin, a synthetic hexapeptide, exerts cyto-protective effects at the mitochondrial level in cardiac and skeletal muscles, both in vitro and in vivo, may also have important neuroprotective bioactivities. This study examined the inhibitory effects of hexarelin on hydrogen peroxide (H2O2)-induced apoptosis in Neuro-2A cells. Neuro-2A cells were treated for 24 h with various concentrations of H2O2 or with the combination of H2O2 and hexarelin following which cell viability and nitrite (NO2−) release were measured. Cell morphology was also documented throughout and changes arising were quantified using Image J skeleton and fractal analysis procedures. Apoptotic responses were evaluated by Real-Time PCR (caspase-3, caspase-7, Bax, and Bcl-2 mRNA levels) and Western Blot (cleaved caspase-3, cleaved caspase-7, MAPK, and Akt). Our results indicate that hexarelin effectively antagonized H2O2-induced damage to Neuro-2A cells thereby (i) improving cell viability, (ii) reducing NO2− release and (iii) restoring normal morphologies. Hexarelin treatment also reduced mRNA levels of caspase-3 and its activation, and modulated mRNA levels of the BCL-2 family. Moreover, hexarelin inhibited MAPKs phosphorylation and increased p-Akt protein expression. In conclusion, our results demonstrate neuroprotective and anti-apoptotic effects of hexarelin, suggesting that new analogues could be developed for their neuroprotective effects.

Introduction: Acute respiratory distress syndrome (ARDS) is an acute form of diffuse lung injury characterized by (i) an intense inflammatory response, (ii) increased pulmonary vascular permeability, and (iii) the loss of respiratory pulmonary tissue. In this article we explore the therapeutic potential of hexarelin, a synthetic hexapeptide growth hormone secretagogue (GHS), in an experimental model of ARDS. Hexarelin has anti-inflammatory properties and demonstrates cardiovascular-protective activities including the inhibition of cardiomyocyte apoptosis and cardiac fibrosis, both of which may involve the angiotensin-converting enzyme (ACE) system. Methods: In our experimental model, ARDS was induced by the instillation of 100 mM HCl into the right bronchus; these mice were treated with hexarelin (320 μg/kg, ip) before (Pre) or after (Post) HCl challenge, or with vehicle. Respiratory system compliance, blood gas analysis, and differential cell counts in a selective bronchoalveolar lavage (BAL) were determined 6 or 24 hours after HCl instillation. In an extended study, mice were observed for a subsequent 14 days in order to assess lung fibrosis. Results: Hexarelin induced a significant improvement in lung compliance and a reduction of the number of total immune cells in BAL 24 hours after HCl instillation, accompanied with a lower recruitment of neutrophils compared with the vehicle group. At day 14, hexarelin-treated mice presented with less pulmonary collagen deposition compared with vehicle-treated controls. Conclusions: Our data suggest that hexarelin can inhibit the early phase of the inflammatory response in a murine model of HCl-induced ARDS, thereby blunting lung remodeling processes and fibrotic development.

L’Italia è uno tra i paesi in Europa e nel mondo dove l’impresa familiare è maggiormente presente nei diversi settori produttivi, con una stima che si avvicina a circa l’85% delle imprese italiane: si tratta di imprese spesso molto antiche, che crescono più di altre tipologie di impresa, hanno una redditività più elevata, e sono fortemente orientate all’espansione nei mercati internazionali (Aidaf, 2019). Molte di queste imprese operano nei settori del Made in Italy, dove i valori dell’autenticità, unicità, qualità, artigianalità, fiducia ed etica, sono spesso associati alla dimensione di impresa familiare (Carcano, 2013) e possono costituire elementi di forte differenziazione. Ciò avviene soprattutto nei settori dell’alimentare, dell’abbigliamento e dell’arredamento, dove è fondamentale investire sulla creazione del valore del brand e sulla sua percezione positiva non solo in Italia ma soprattutto all’estero (Alon et al., 2016). Per analizzare i diversi approcci alla comunicazione dei valori familiari in un contesto internazionale, abbiamo dunque individuato i valori tipicamente associabili alle imprese familiari evidenziati dalla letteratura, che ci hanno consentito di sviluppare uno schema di analisi della comunicazione estera. Successivamente, è stata condotta un’analisi esplorativa sui siti web di un campione di novanta imprese italiane appartenenti ai settori del made in Italy sopra menzionati.

VGF gene encodes for a neuropeptide precursor of 68 kDa composed by 615 (human) and 617 (rat, mice) residues, expressed prevalently in the central nervous system (CNS), but also in the peripheral nervous system (PNS) and in various endocrine cells. This precursor undergoes proteolytic cleavage, generating a family of peptides different in length and biological activity. Among them, TLQP-21, a peptide of 21 amino acids, has been widely investigated for its relevant endocrine and extraendocrine activities. The complement complement C3a receptor-1 (C3aR1) has been suggested as the TLQP-21 receptor and, in different cell lines, its activation by TLQP-21 induces an increase of intracellular Ca2+. This effect relies both on Ca2+ release from the endoplasmic reticulum (ER) and extracellular Ca2+ entry. The latter depends on stromal interaction molecules (STIM)-Orai1 interaction or transient receptor potential channel (TRPC) involvement. After Ca2+ entry, the activation of outward K+-Ca2+-dependent currents, mainly the KCa3.1 currents, provides a membrane polarizing influence which offset the depolarizing action of Ca2+ elevation and indirectly maintains the driving force for optimal Ca2+ increase in the cytosol. In this review, we address the main endocrine and extraendocrine actions displayed by TLQP-21, highlighting recent findings on its mechanism of action and its potential in different pathological conditions.

The critical role of neuroinflammation in favoring and accelerating the pathogenic process in Alzheimer's disease (AD) increased the need to target the cerebral innate immune cells as a potential therapeutic strategy to slow down the disease progression. In this scenario, mesenchymal stem cells (MSCs) have risen considerable interest thanks to their immunomodulatory properties, which have been largely ascribed to the release of extracellular vesicles (EVs), namely exosomes and microvesicles. Indeed, the beneficial effects of MSC‐EVs in regulating the inflammatory response have been reported in different AD mouse models, upon chronic intravenous or intracerebroventricular administration. In this study, we use the triple‐transgenic 3xTg mice showing for the first time that the intranasal route of administration of EVs, derived from cytokine‐preconditioned MSCs, was able to induce immunomodulatory and neuroprotective effects in AD. MSC‐EVs reached the brain, where they dampened the activation of microglia cells and increased dendritic spine density. MSC‐EVs polarized in vitro murine primary microglia toward an anti‐inflammatory phenotype suggesting that the neuroprotective effects observed in transgenic mice could result from a positive modulation of the inflammatory status. The possibility to administer MSC‐EVs through a noninvasive route and the demonstration of their anti‐inflammatory efficacy might accelerate the chance of a translational exploitation of MSC‐EVs in AD., In a preclinical model of Alzheimer's disease, characterized by neuronal damage and a high rate of inflammation (left), the intranasal (IN) administration of extracellular vesicles (EVs) derived from mesenchymal stromal/stem cells (MSCs) operates in dampening inflammation (by reducing microglia activation) and in inducing neuroprotective effects (by decreasing spine loss) (right). These data suggest the possibility that the IN route administration of MSC‐EVs might accelerate the chance of a translational exploitation of MSC‐EVs toward therapy.

Neurodegenerative diseases, including Alzheimer disease (AD), Parkinson disease (PD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and Huntington disease, are characterized by the loss of neurons as well as neuronal function in multiple regions of the central and peripheral nervous systems. Several studies in animal models have shown that androgens have neuroprotective effects in the brain and stimulate axonal regeneration. The presence of neuronal androgen receptors in the peripheral and central nervous system suggests that androgen therapy might be useful in the treatment of neurodegenerative diseases. To illustrate, androgen therapy reduced inflammation, amyloid-β deposition, and cognitive impairment in patients with AD. As well, improvements in remyelination in MS have been reported; by comparison, only variable results are observed in androgen treatment of PD. In ALS, androgen administration stimulated motoneuron recovery from progressive damage and regenerated both axons and dendrites. Only a few clinical studies are available in human individuals despite the safety and low cost of androgen therapy. Clinical evaluations of the effects of androgen therapy on these devastating diseases using large populations of patients are strongly needed.

In several neurodegenerative diseases, including amyotrophic lateral sclerosis, alterations of redox homeostasis within the cell lead to oxidative stress, abnormalities in mitochondrial morphology and biochemistry, damage to DNA, RNA, proteins and lipids, and ultimately to apoptosis. Intracellular hydrogen peroxide (H2O2), generated through oxidative stress, activates the apoptotic pathway by the phosphorylation of the pro-apoptotic regulator Bax and the downregulation of the anti-apoptotic protein Bcl-2, with the subsequent activation of caspases – 3 and – 7. Consequently, phosphoinositide 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) are modulated, supporting cell survival or inducing cell death. Hexarelin is a well-known growth hormone secretagogue endowed with growth hormone-releasing effects and neuroprotective activities, such as prevention of status epilepticus and promotion of neurogenesis. Moreover, hexarelin reduced the activation of caspase – 3 and caspase – 7 caused by brain hypoxia-ischemia procedures in neonatal rats, and exerted protective effects against β-amyloid cytotoxicity in vitro. In this study, we explored the protective effects of hexarelin against oxidative stress and its anti-apoptotic mechanism to attenuate H2O2-induced neurotoxicity in Neuro-2A cells, a mouse neuroblastoma cell line. Neuro-2A cells were incubated with increasing concentration of H2O2 for 24 h. MTT assay was used to measure changes in the viability of cells, and 100 µM H2O2 was selected as the lower concentration that significantly induced a reduction of cells survival. Further measurements by real-time PCR analysis showed that 24 hours treatment with 100 µM H2O2 induced significant changes in caspases – 3 and – 7, Bax and Bcl-2 mRNA levels, inducing cell apoptosis. Hexarelin blunted H2O2 effects both increasing cell viability and by opposing changes in caspases – 3 and – 7, Bax and Bcl-2 mRNA levels. Western blot analysis confirmed that hexarelin activate the Akt, p38 and ERK1/2 pathways. In conclusion, our results suggest that hexarelin exerts neuroprotective activities against H2O2-induced toxicity in Neuro-2A cells. Further experiments are needed to identify the precise molecular mechanisms underlying this neuroprotective effects and to clarify whether other GHS moieties exert the same neuroprotective effects.

Combined AntiRetroviral Treatments (cARTs) used for HIV infection may result in varied metabolic complications, which in some cases, may be related to patient genetic factors, particularly microRNAs. The use of monozygotic twins, differing only for HIV infection, presents a unique and powerful model for the controlled analysis of potential alterations of miRNAs regulation consequent to cART treatment. Profiling of 2578 mature miRNA in the subcutaneous (SC) adipose tissue and plasma of monozygotic twins was investigated by the GeneChip® miRNA 4.1 array. Real-time PCR and ddPCR experiments were performed in order to validate differentially expressed miRNAs. Target genes of deregulated miRNAs were predicted by the miRDB database (prediction score > 70) and enrichment analysis was carried out with g:Profiler. Processes in SC adipose tissue most greatly affected by miRNA up-regulation included (i) macromolecular metabolic processes, (ii) regulation of neurogenesis, and (iii) protein phosphorylation. Furthermore, KEGG analysis revealed miRNA up-regulation involvement in (i) insulin signaling pathways, (ii) neurotrophin signaling pathways, and (iii) pancreatic cancer. By contrast, miRNA up-regulation in plasma was involved in (i) melanoma, (ii) p53 signaling pathways, and (iii) focal adhesion. Our findings suggest a mechanism that may increase the predisposition of HIV+ patients to insulin resistance and cancer.

Androgens in women, as well as in men, are intrinsic to maintenance of (i) reproductive competency, (ii) cardiac health, (iii) appropriate bone remodeling and mass retention, (iii) muscle tone and mass, and (iv) brain function, in part, through their mitigation of neurodegenerative disease effects. In recognition of the pluripotency of endogenous androgens, exogenous androgens, and selected congeners, have been prescribed off-label for several decades to treat low libido and sexual dysfunction in menopausal women, as well as, to improve physical performance. However, long-term safety and efficacy of androgen administration has yet to be fully elucidated. Side effects often observed include (i) hirsutism, (ii) acne, (iii) deepening of the voice, and (iv) weight gain but are associated most frequently with supra-physiological doses. By contrast, short-term clinical trials suggest that the use of low-dose testosterone therapy in women appears to be effective, safe and economical. There are, however, few clinical studies, which have focused on effects of androgen therapy on pre- and post-menopausal women; moreover, androgen mechanisms of action have not yet been thoroughly explained in these subjects. This review considers clinical effects of androgens on women's health in order to prevent chronic diseases and reduce cancer risk in gynecological tissues.

TLQP-21, a peptide encoded by the prohormone VGF, is expressed in neuroendocrine cells and can modulate inflammatory processes. Since TLQP-21 can bind the complement 3a receptor 1 on macrophages, interest has risen in this peptide as a potential drug for the treatment of Acute Respiratory Distress Syndrome (ARDS), whose hospital mortality can reach 35–46%. Since no effective pharmacologic therapies are available, our aim was to exploit the potential of a short analog of TLQP-21(JMV5656) in order to modulate the inflammatory process in ARDS and the progression to pulmonary fibrosis in an experimental model of unilateral acid aspiration in mice. Mice were divided in 2 treatment groups. In the acute protocol, mice received intra-peritoneal injection of either vehicle or 0.6 mg/kg JMV5656 on experimental days 1 and 2, and ARDS was induced on day 3 under deep anesthesia by instillation of HCl (1.5 ml/kg of 0.1 M HCl in 0.9% NaCl) into the right lung; all measurements were performed 24 h later. In the subacute protocol, mice were treated as previously, but treatment with vehicle or JMV5656 was repeated also on day 4 and measurements were made 2 weeks later. Twenty-four hours after acid instillation, the total number of immune cell in the BAL rose sharply due primarily to an increase in the PMN population which increased from 1% up to 58% of total cell numbers. JMV5656 significantly reduced PMN recruitment into the alveolar space, but had no effects on cytokine levels in BAL. Two weeks after acid injury, static compliance of the right lung was significantly higher in the JMV5656-treated group compared to vehicle-treated group. Treatment with JMV5656 also blunted the acid-induced collagen deposition in the right lung. These results suggest that JMV5656 can ameliorate mechanical compliance, and reduce collagen deposition in acid-injured lungs in mice. This effect was likely due to the ability of JMV5656 to inhibit PMN recruitment in the injured lung.

Growth hormone secretagogues (GHS) are a family of synthetic molecules, first discovered in the late 1970s for their ability to stimulate growth hormone (GH) release. Many effects of GHS are mediated by binding to GHS-R1a, the receptor for the endogenous hormone ghrelin, a 28-amino acid peptide isolated from the stomach. Besides endocrine functions, both ghrelin and GHS are endowed with some relevant extraendocrine properties, including stimulation of food intake, anticonvulsant and anti-inflammatory effects, and protection of muscle tissue in different pathological conditions. In particular, ghrelin and GHS inhibit cardiomyocyte and endothelial cell apoptosis and improve cardiac left ventricular function during ischemia−reperfusion injury. Moreover, in a model of cisplatin-induced cachexia, GHS protect skeletal muscle from mitochondrial damage and improve lean mass recovery. Most of these effects are mediated by GHS ability to preserve intracellular Ca2+ homeostasis. In this review, we address the muscle-specific protective effects of GHS mediated by Ca2+ regulation, but also highlight recent findings of their therapeutic potential in pathological conditions characterized by skeletal or cardiac muscle impairment.

Background Ventilator-induced diaphragmatic dysfunction (VIDD) is a common event during mechanical ventilation (MV) leading to rapid muscular atrophy and contractile dysfunction. Recent data show that renin-angiotensin system is involved in diaphragmatic skeletal muscle atrophy after MV. In particular, angiotensin-II can induce marked diaphragm muscle wasting, whereas angiotensin-(1–7) (Ang-(1–7)) could counteract this activity. This study was designed to evaluate the effects of the treatment with Ang-(1–7) in a rat model of VIDD with neuromuscular blocking agent infusion. Moreover, we studied whether the administration of A-779, an antagonist of Ang-(1–7) receptor (Mas), alone or in combination with PD123319, an antagonist of AT2 receptor, could antagonize the effects of Ang-(1–7). Methods Sprague-Dawley rats underwent prolonged MV (8 h), while receiving an iv infusion of sterile saline 0.9% (vehicle) or Ang-(1–7) or Ang-(1–7) + A-779 or Ang-(1–7) + A-779 + PD123319. Diaphragms were collected for ex vivo contractility measurement (with electric stimulation), histological analysis, quantitative real-time PCR, and Western blot analysis. Results MV resulted in a significant reduction of diaphragmatic contractility in all groups of treatment. Ang-(1–7)-treated rats showed higher muscular fibers cross-sectional area and lower atrogin-1 and myogenin mRNA levels, compared to vehicle treatment. Treatment with the antagonists of Mas and Ang-II receptor 2 (AT2R) caused a significant reduction of muscular contractility and an increase of atrogin-1 and MuRF-1 mRNA levels, not affecting the cross-sectional fiber area and myogenin mRNA levels. Conclusions Systemic Ang-(1–7) administration during MV exerts a protective role on the muscular fibers of the diaphragm preserving muscular fibers anatomy, and reducing atrophy. The involvement of Mas and AT2R in the mechanism of action of Ang-(1–7) still remains controversial.

Intracellular hydrogen peroxide (H2O2) generated through oxidative stress is involved in necrosis and apoptosis, ultimately resulting in aging, cancer and several neurodegenerative diseases, including Amyotrophic Lateral Sclerosis (ALS). Evidences demonstrate that neurotrophic factors, including insulin-like growth factor 1 (IGF-1), regulate survival and differentiation of nerve cells and maintain neuronal structural integrity. However, given the high tumorigenic potential of exogenous IGF-1 administration, alternative methods to increase endogenous IGF-1 expression should be develop. Growth hormone secretagogues (GHS) are a family of peptides that stimulates the secretion of growth hormone and IGF-1, and exerts neuroprotective effects against β-amyloid cytotoxicity. Among them, hexarelin has shown growth hormone-releasing effects and neuroprotective activities, such as prevention of status epilepticus and promotion of neurogenesis. Moreover, hexarelin reduces the activation of caspase-3 and caspase-7 involved in apoptosis pathway, caused by brain hypoxia-ischemia in neonatal rats. In this study, we explored the protective effects of hexarelin against oxidative stress and its anti-apoptotic mechanisms to attenuate H2O2-induced neurotoxicity in mouse neuroblastoma Neuro2A cells. Neuro2A cells were incubated with increasing concentration of H2O2 for 24 h. After determining the optimal concentration of H2O2, Neuro2A cells were treated with H2O2 in absence or presence of hexarelin (24 h). Cell viability was evaluated using MTT assay, and mRNA levels of Bax, Bcl-2, caspase-3 and caspase-7 were obtained by Real Time PCR. Western blot analysis was used to measure ERK1/2, Akt and NF- κB proteins expression. In conclusion, our results suggest that hexarelin exerts neuroprotective activities against H2O2-induced toxicity in Neuro2A cells. Further experiments are needed to identify the molecular mechanisms underlying this neuroprotective effects.

The nucleophosmin 1 gene (NPM1) is the most frequently mutated gene in acute myeloid leukemia. Notably, NPM1 mutations are always accompanied by additional mutations such as those in cohesin genes RAD21, SMC1A, SMC3, and STAG2 but not in the cohesin regulator, nipped B-like (NIPBL). In this work, we analyzed a cohort of adult patients with acute myeloid leukemia and NPM1 mutation and observed a specific reduction in the expression of NIPBL but not in other cohesin genes. In our zebrafish model, overexpression of the mutated form of NPM1 also induced downregulation of nipblb, the zebrafish ortholog of human NIPBL. To investigate the hematopoietic phenotype and the interaction between mutated NPM1 and nipblb, we generated a zebrafish model with nipblb downregulation which showed an increased number of myeloid progenitors. This phenotype was due to hyper-activation of the canonical Wnt pathway: myeloid cells blocked in an undifferentiated state could be rescued when the Wnt pathway was inhibited by dkk1b mRNA injection or indomethacin administration. Our results reveal, for the first time, a role for NIPBL during zebrafish hematopoiesis and suggest that an interplay between NIPBL/NPM1 may regulate myeloid differentiation in zebrafish and humans through the canonical Wnt pathway and that dysregulation of these interactions may drive leukemic transformation.

Hexarelin is a well-known Growth Hormone Secretagogues (GHS) that possesses growth hormone-releasing effects and neuroprotective activities, such as prevention of status epilepticus and promotion of neurogenesis. Moreover, hexarelin reduces the activation of caspase 3 and 7 caused by brain hypoxia-ischemia in neonatal and adult rats and exerts protective effects against β-amyloid cytotoxicity. Several neurodegenerative diseases (NDDs), such as amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Huntington’s disease (HD), are characterized by an increased production of reactive oxygen species (ROS), which lead to a dysregulation of mitochondrial dynamics, neuronal plasticity and apoptosis. In particular, ROS cause the downregulation of the anti-apoptotic protein Bcl-2 and the phosphorylation of the pro-apoptotic molecule Bax, with the subsequent activation of caspases 3 and 7. In this study, we explored the protective effects of hexarelin against oxidative stress and its anti-apoptotic mechanisms to attenuate H2O2-induced neurotoxicity using mouse neuroblastoma Neuro2A cells. MTT assay was used to measure changes in the viability of Neuro2A cells at different concentrations of H2O2 for 24 hours, and 100 μM H2O2 was selected as the lower concentration that significantly induced oxidative stress and a reduction of cells survival. Further assays showed that 24 hours treatment with 100 μM H2O2 induced significant changes in caspase 3 and caspase 7 mRNA levels and induced apoptosis. Treatment of Neuro2A cells with hexarelin blunted 100 μM H2O2 effects in MTT assay and cytotoxicity assay (CCK8). The apoptosis mechanism induced by H2O2 was reduced by hexarelin, which was confirmed by real-time PCR analysis of caspase 3, caspase 7, Bcl-2 and Bax mRNA expression. In conclusion, our results suggest that hexarelin exerts neuroprotective activities against H2O2-induced toxicity in Neuro2A cells. Further experiments are needed to identify the molecular mechanisms underlying the neuroprotective effects.

Introduction: Intracellular hydrogen peroxide (H2O2) generated through oxidative stress is involved in necrosis and apoptosis, ultimately resulting in aging, cancer and several neurodegenerative diseases, including Amyotrophic Lateral Sclerosis (ALS). In particular reactive oxygen species (ROS) modulate the extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase (PI3K/Akt) pathways, and the activation of pro-apoptotic protein Bax. Evidences demonstrate that neurotrophic factors, including insulin-like growth factor 1 (IGF-1), regulate survival and differentiation of nerve cells and maintain neuronal structural integrity. Experimental data showed that growth hormone secretagogues (GHS) not only inhibit cytotoxic effect of β-amyloid in N9 cells and counteract atrophy in cachectic rats, but can stimulate the release of IGF-1. Hexarelin is a well-known GHS endowed with growth hormone-releasing effects and neuroprotective activities, such as prevention of status epilepticus and promotion of neurogenesis. Moreover, hexarelin reduces the activation of caspase-3 and caspase-7 involved in apoptosis pathway, caused by brain hypoxia-ischemia in neonatal rats. In this study, we explored the protective effects of hexarelin against oxidative stress and its anti-apoptotic mechanisms to attenuate H2O2-induced neurotoxicity in mouse neuroblastoma Neuro2A cells. Materials and methods: Neuro2A cells were incubated with increasing concentration of H2O2 for 24 h. After determining the optimal concentration of H2O2, Neuro2A cells were treated with H2O2 in absence or presence of hexarelin (24 h). Cell viability was evaluated using MTT assay, and mRNA levels of Bax, Bcl-2, caspase-3 and caspase-7 were obtained by Real Time PCR. Western blot analysis was used to measure ERK1/2, Akt and NF- κB proteins expression. Results: H2O2 100 μM was selected as the lower concentration that significantly induced oxidative stress and a reduction of cells viability. MTT assay demonstrated that hexarelin increased cells survival. H2O2 induced the activation of apoptosis pathway: the mRNA level of pro-apoptotic protein Bax was significantly increased as well as the levels of caspase-3 and caspase-7, while Bcl-2 mRNA levels were reduced. Hexarelin also reduced the apoptosis activation: Bax, caspase-3 and caspase-7 mRNA levels were reduced while anti-apoptotic Bcl-2 mRNA levels were increased. Western blotting analysis confirmed the role of hexarelin via the anti-apoptotic Akt and ERK1/2 pathways and the reduction of NF-κB translocation to the nucleus. Discussion and conclusion: In conclusion, our results suggest that hexarelin exerts neuroprotective activities against H2O2-induced toxicity in Neuro2A cells. Further experiments are needed to identify the molecular mechanisms underlying this neuroprotective effects.

Background: Metabolic complications represent a common and serious problem associated with HIV infection and combined Antiretroviral Therapy (cART). Alterations in body fat distribution are associated with significantly increased risks of (i) metabolic derangements, (ii) cardiovascular pathologies, and (iii) insulin resistance. A case control study showed that in subcutaneous adipose tissue from HIV-infected patients on cART presenting lipodystrophy (LS), the levels of miRNA-218 were upregulated and those of lipin-1, a putative target gene of miRNA-218, were downregulated compared with HIV-negative subjects. Lipin-1 is one of the most important factors linked to development of LS. Lipin-1, by controlling PPARγ2, regulates the expression of specific genes, such as that of glucose transporter type 4 (GLUT-4), required for maturation and maintenance of adipocytes. Objectives: To determine whether lopinavir/ritonavir (LPV/RTV) can modulate lipogenesis in adipocytes affecting miRNA-218 and lipin-1 mRNA expression, and to investigate the functional link between miRNA-218 and GLUT-4 mRNA expression. Methods: Differentiated 3T3-L1 cells were treated with various combinations of LPV/RTV, followed by measurements of cell viability, lipid accumulation, lipin-1 and GLUT-4 mRNA and miRNA-218 levels. Transfection of anti-miR-218 or a miRNA-218 mimic were used to investigate the role of miRNA-218 in lipogenesis. Results: LPV/RTV treatment of 3T3-L1 cells did not affect the viability of differentiated 3T3-L1 cells, but caused (i) a significant decrease of lipid accumulation, (ii) an overexpression of miRNA-218, and (iii) a reduction of lipin-1 and GLUT-4 mRNA levels. The anti-miR-218 transfection of 3T3-L1 cells significantly ameliorated the adipogenic dysfunction and restored mRNA levels of lipin-1 and GLUT-4 consequent to LPV/RTV treatment. By contrast, 3T3-L1 cells transfected with a specific miRNA-218 mimic showed (i) an overexpression of miRNA-218, (ii) a reduced cellular lipid fraction, and (iii) decreased levels of mRNA for lipin-1 and GLUT-4. Conclusion: 3T3-L1 cells, treated with LPV/RTV, show altered lipid content due to increased miRNA-218 levels, which affects lipin-1 mRNA. Moreover, increased miRNA-218 levels were inversely correlated with changes in GLUT-4 expression, which suggests a role for miRNA-218 in mediating the insulin resistance consequent to cART.