Modulation of Myotilin and Fylamin C in Various Muscle Diseases: A Microarray Analysis

COMPARTMENT to identify the localization in cells. We showed that both MYOT and FLNC were significantly modulated in various muscle diseases. In particular, MYOT and FLNC mRNA were significantly downregulated in Acute quadriplegic myopathy (AQM) and Amyotrophic Lateral Sclerosis (ALS) compared to normal human skeletal muscle. Furthermore, the GIANT analysis showed a relationship confidence of 0.23 to MYOT and FLNC, confirming their strong correlation. These data provide to support our hypothesis that a positive correlation exists between MYOT and FLNC. Larger studies are needed to evaluate if MYOT and FLNC may be a promising clinical biomarker in subjects with diseases of the muscle

HIV RNA Suppression and Immune Restoration: Can We Do Better?

HAART has significantly changed the natural history of HIV infection: patients receiving antiretrovirals are usually able to control viremia, even though not all virological responders adequately recover their CD4+ count. The reasons for poor immune restoration are only partially known and they include genetic, demographic and immunologic factors. A crucial element affecting immune recovery is immune activation, related to residual viremia; indeed, a suboptimal virological control (i.e., low levels of plasma HIV RNA) has been related with higher levels of chronic inflammation and all-cause mortality. The sources of residual viremia are not yet completely known, even though the most important one is represented by latently infected cells. Several methods, including 2-LTR HIV DNA and unspliced HIV RNA measurement, have been developed to estimate residual viremia and predict the outcome of antiretroviral therapy. Considering that poor immunologic responders are exposed to a higher risk of both AIDS-related and non-AIDS-related diseases, there is a need of new therapeutic strategies, including immunomodulators and drugs targeting the latent viral reservoirs, in order to face residual viremia but also to “drive” the host immunologic responses

Heme oxygenase-1 in central nervous system malignancies

Central nervous system tumors are the most common pediatric solid tumors and account for 20%-25% of all childhood malignancies. Several lines of evidence suggest that brain tumors show altered redox homeostasis that triggers the activation of various survival pathways, leading to disease progression and chemoresistance. Among these pathways, heme oxygenase-1 (HO-1) plays an important role. HO-1 catalyzes the enzymatic degradation of heme with the simultaneous release of carbon monoxide (CO), ferrous iron (Fe2+), and biliverdin. The biological effects of HO-1 in tumor cells have been shown to be cell-specific since, in some tumors, its upregulation promotes cell cycle arrest and cellular death, whereas, in other neoplasms, it is associated with tumor survival and progression. This review focuses on the role of HO-1 in central nervous system malignancies and the possibility of exploiting such a target to improve the outcome of well-established therapeutic regimens. Finally, several studies show that HO-1 overexpression is involved in the development and resistance of brain tumors to chemotherapy and radiotherapy, suggesting the use of HO-1 as an innovative therapeutic target to overcome drug resistance. The following keywords were used to search the literature related to this topic: nuclear factor erythroid 2 p45-related factor 2, heme oxygenase, neuroblastoma, medulloblastoma, meningioma, astrocytoma, oligodendroglioma, glioblastoma multiforme, and gliomas

Identification of Novel Markers of Prostate Cancer Progression, Potentially Modulated by Vitamin D

Prostate cancer (PCa) is one of the most common cancers in men. The main risk factors associated with the disease include older age, family history of the disease, smoking, alcohol and race. Vitamin D is a pleiotropic hormone whose low levels are associated with several diseases and a risk of cancer. Here, we undertook microarray analysis in order to identify the genes involved in PCa. We analyzed three PCa microarray datasets, overlapped all genes significantly up-regulated, and subsequently intersected the common genes identified with the down-regulated genes transcriptome of LNCaP cells treated with 1α,25(OH)2D3, in order to identify the common genes involved in PCa and potentially modulated by Vitamin D. The analysis yielded 43 genes potentially involved in PCa and significantly modulated by Vitamin D. Noteworthy, our analysis showed that six genes (PRSS8, SOX4, SMYD2, MCCC2, CCNG2 and CD2AP) were significantly modulated. A Pearson correlation analysis showed that five genes out of six (SOX4 was independent), were statistically correlated with the gene expression levels of KLK3, and with the tumor percentage. From the outcome of our investigation, it is possible to conclude that the genes identified by our analysis are associated with the PCa and are potentially modulated by the Vitamin D

One Year of COVID-19 Pandemic in Italy: Effect of Sedentary Behavior on Physical Activity Levels and Musculoskeletal Pain among University Students

: The COVID-19 outbreak forced Italian students to reduce their daily activities, inducing a seden-tary attitude that was worsened by distanced learning. This study aimed to survey the physical activity levels that were maintained before and during the social restrictions following the pan-demic, their correlation to musculoskeletal pain, as well as analyzing the impact of these COVID-19 restrictions on pain and fatigue that affects daily life activities. A total of 2044 students completed the online questionnaire, of which the results of 1654 participants were eligible. Before the pandemic, the levels of physical activity were distributed as: 19.9% no activity, 30.1% light ac-tivity, 21.5% moderate activity, and 28.5% high activity. After one year of the pandemic, 30.6% of the participants were inactive, 48.1%, 10.9%, and 10.5% stated as maintaining, respectively, light, moderate and high levels of physical activity. Furthermore, 43.5% reported neck pain and 33.5% stated to experience low back pain. Physical activity levels lower than 150 min/week may have predisposed students to suffer from neck pain (1.95 OR at 95% CI, 1.44-2.64) and low back pain (1.79 OR at 95% CI, 1.29-2.49). A positive correlation between physical activity levels, Verbal Descriptive Scale (VDS), and pain frequency have been observed for neck and low back pain (p-value < 0.05). Finally, low physical activity levels were associated with musculoskeletal pain onset and pain worsening

Bortezomib modulates CHIT1 and YKL40 in monocyte-derived osteoclast and in myeloma cells

Osteolytic bone disease is a common manifestation of multiple myeloma (MM) that leads to progressive skeleton destruction and is the most severe cause of morbidity in MM patients.It results from increased osteolytic activity and decrease osteoblastic function. Activation of mammalian chitinases CHIT1 and YKL40 is associated with osteoclast (OCs) differentiation and bone digestion. In the current study, we investigated the effect of two Bortezomib’s concentration (BO) (2.5 nM and 5nM) on osteoclastogenesis by analyzing regulation of chitinase expression. OCs exposition to BO was able to inhibit the expression of different OCs markers such as RANK, CTSK, TRAP and MMP9. In addition BO-treatment reduced CHIT1 enzymatic activity and both CHIT1 and YKL40 mRNA expression levels and cytoplasmatic and secreted protein. Moreover, immunofluorescence evaluation of mature OCs showed that BO was able to translocate YKL40 into the nucleus, while CHIT1 remained into the cytoplasm. Since MM cell lines such as U266, SKM-M1 and MM1 showed high levels of CHIT1 activity, we analyzed bone resorption ability of U266 using dentin disc assay resorption pits. Silencing chitinase proteins in U266 cell line with specific siRNAs, resulted in pits number reduction on dentine discs. In conclusion, we showed that BO decreases osteoclastogenesis and reduces bone resorption in OCs and U266 cell line by modulating the chitinases CHIT1 and YKL40. These results indicate that chitinases may be a therapeutic target for bone disease in MM patients

Chitotriosidase expression in monocyte-derived dendritic cells

Chitotriosidase (CHIT1) belongs to 18 glycosyl-hydrolase family, an ancient gene family that is widely expressed from prokaryotes to eukaryotes [1]. CHIT1 is a very critical enzyme to regulate the susceptibility to infection of organisms containing chitin as structural components. Conversely, during the development of acute/chronic inflammatory disorders, the enzymatic activity of CHIT1 increases significantly. The CHIT1 is expressed in activated macrophages as well in different lines monocyte-derived such as Kupffer cells and osteoclasts [2]. So far, it is unknown whether CHIT1 is expressed in other cells involved in the immune response such as monocyte-derived DCs. In this study we have investigated whether CHIT1 is produced in monocyte-derived DCs (moDCs) and the differential expression of CHIT1 during the different stage of moDCs differentiation. The presence of CHIT1 were examined by real time RT-PCR, Western Blot and Confocal Immunofluorescence, in Immature Dendritic cells (iDCs), generated from human monocytes by stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) and in mature Dendritic cells (mDCs), obtained by using lipopolysaccharide (LPS) and interferon-gamma (IFN-g). We observed that CHIT1 was expressed during the DCs differentiation and maturation process in time dependent manner. The maturation of DCs showed a significantly increased expression of CHIT1 mRNA and protein. Furthermore, the CHIT1 was evenly distributed in cytoplasm both in iDCs and in mDCs. The enzymatic activity confirmed that CHIT1 could play a role in moDCs function. Taken together, our data confirm the crucial role of CHIT1 in primary immune responses and indicate that could be correlated with the immunogenicity of DCs

Mesenchymal stem cell-based tissue engineering strategy for cartilage regeneration: A morphomolecular study

Articular cartilage is an avascular and aneural tissue with poor self-repair capacity. Pathological conditions leading to the cartilage degeneration, such as osteoarthritis (OA), have prompted the development of strategies aimed to its regeneration, such as mesenchymal stem cells (MSCs)-based tissue engineering approach. The aim of this study was to investigate if chondrocytes, differentiated from rat adipose tissue derived-MSCs (AMSCs) and seeded on Collagen Cell Carrier (CCC) scaffolds, are able to constitute a morphologically and biochemically healthy hyaline cartilage. To this purpose the AMSCs were primarily differentiated in chondrocytes through chondrogenic medium and subsequently cultured for 6 weeks on CCC scaffolds. The expression of osteoblast (Runt-related transcription factor 2 (RUNX2) and osteocalcin), chondrocyte (collagen I, II and lubricin) and apoptosis (caspase-3) biomarkers were evaluated in undifferentiated AMSCs, AMSCs-derived chondrocytes cultured in monolayer and AMSCs-derived chondrocytes seeded on CCC scaffolds, by different techniques such as immunohistochemistry, ELISA, Western blot and gene expression analyses. AMSCs-derived chondrocytes cultured on CCC scaffolds showed the increased expression of collagen II and lubricin, whereas the expression of collagen I, RUNX2, osteocalcin and caspase-3 resulted decreased when compared to the other groups. In conclusion, the results of this study suggest a possible role of AMSCs and the use of CCC scaffolds for therapeutic strategies aimed to the articular cartilage regeneration

Expression and localization of CHI3L1 in monocyte derived dendritic cells

Chitinase-3-like-1 protein (CHI3L1) also called YKL-40, is a 40 kDa mammalian glycoprotein which is a heparin, chitin and collagen binding member of the mammalian chitinase-like proteins. Biological activities of CHI3L1 embrace regulation of cell proliferation, adhesion, angiogenesis, migration and activation. CHI3L1 is produced by variety of cells, including neutrophils, monocytes/macrophages, osteoclasts and Kupffer cells [1]. CHI3L1 secretion is induced by interferon (INF)-g and interleukin (IL)-6 and is an acute phase reactant associated with disease severity and mortality in a variety of infectious [2]. In this study, we have examined the expression and localization of CHI3L1 during the differentiation and maturation of monocyte derived dendritic cells by real time RT-PCR, Western Blot, Confocal Immunofluorescence, and Immunocytochemical assays. Potential nuclear localization signal (NLS) was determinated using the open source software cNLS Mapper and Chimera. Peripheral blood monocytes were differentiated toward immature DCs (iDC) and mature DCs (mDCs) through a combination of factors and cytokines. Our result showed, for the first time, that CHI3L1 is expressed during the process of differentiation and maturation of DCs in time dependent manner. Furthermore, CHI3L1 is evenly distributed in cytoplasm and in the nucleus of both the iDCs and mDCs. In conclusion, the discovery of CHI3L1 expression in DCs has opened new dilemma for designing DC-based cancer immunotherapeutic. In fact, on the light of these results one can’t exclude that as well as activated Tumor-associated macrophages (TAMs) also DCs infiltration could to be a significant unfavorable prognostic factor for cancer patients

Unexpected ovarian activity in premenopausal breast cancer survivors treated with exemestane and GnRH analogues

n/