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Chronic renal failure is a progressive disease characterized by a gradual destruction of nephrons and a consequent reduction of kidney function. End-stage renal disease (ESRD) necessitates renal replacement therapy as peritoneal dialysis, hemodialysis, or transplantation. Patients affected by ESRD or in hemodialysis are at risk for developing a number of comorbidities including hypertension, anemia, risk of bleeding, susceptibility to infection, medication side effects, and oral manifestations associated with the disease itself and with hemodialysis treatment. In this context, oral diseases represent a potential and preventable cause of poor health outcomes in people with ESRD due to their relation to infection, inflammation, and malnutrition. The aim of this article was to review ESRD and hemodialysis-associated manifestations and to describe the dental operative protocols for patients awaiting kidney transplantation in light of the most recent literature.

The Endocannabinoid System (ECS) comprising the CB1 and CB2 receptors and their endogenous ligands is a central signalling system regulating food intake and energy balance. It is also present in peripheral tissues where it is involved in cell proliferation and survival. It has been shown that in colon cancer cells, the CB1 receptor antagonist SR171416 reduces colon cancer cell growth by acting as an inverse agonist rather than an antagonist [1]. Starting from this observation and from evidence indicating that some biological responses to cannabinoids depend on estrogen levels and some selective estrogen receptor modulators can bind the CB1 receptor [2], we aimed to study the effects of the CB1 receptor ligand anandamide (AEA) on colon cancer cell proliferation and its ability to modulate some survival and proliferating pathways including Akt, MAPK/ERK and estrogen receptor (ER) b signalling which is the predominant ER pathway in colonic epithelium. We used an AEA-analogue and a selective inhibitor of fatty acid amide hydrolase (FAAH) that enhances intracellular levels of AEA and studied proliferation and cell cycle progression on human adenocarcinoma cells DLD1 and SW620. Our results showed that increased levels of AEA significantly reduced cell proliferation in both cell lines at 24 and 48 h also inducing an S phase cell cycle accumulation. The AEA-induced inhibition of cell growth was mediated by a reduced expression of phoshoAkt and phosphoERK and, at the same time, by an induction of ERβ expression. These data suggest that AEA can reduces colon cancer cell proliferation by interfering with different signalling pathways., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017

In the search of new compounds useful for the treatment of neurological disorders, medicinal plant research has progressed widely in the last decade, particularly because of the hypothesis that natural extracts possess a low or absent toxicity. Aloysia polystachia (Griseb.) Moldenke belonging to the family of Verbenaceae has been used in the traditional medicine for a variety of indications and as a sedative. Recently, some evidence reported that it exhibits some antidepressant- and anxiolytic-like effects involving the modulation of GABA system in preclinical studies in mice (1). However no investigations have been performed on its (geno)toxicity in long-term studies as it would be required for its use in neurological pathologies. In this study, we investigated the genotoxic potential of a water soluble extract of Aloysia polystachia leaves, administered orally every day for 90 days to 20 New Zealand white rabbits homogeneous for weight and age. Subjects were divided into four groups: the control group, and three experimental groups fed with a diet supplemented with 1g/kg; 1.5 g/kg and 2g/kg of Aloysia Polystachia, then peripheral blood was drawn from the vein auricularis marginalis before and after 45 and 90 days of treatment. Cytogenetic analysis was performed on each subject at each time point and chromosome aberrations (structural and numerical) and mitotic index were used as a measure of DNA damage and cytotoxicity (2). Results indicated that Aloysia polystachia extract induces a significant increase in the percentage of aberrant cells as well as in the aberration frequency (mainly chromatid breaks and fragments) associated with a decreasing trend of the mitotic index. The DNA damage was particularly higher after the first 45 days of treatment whereas it remains significantly high but almost unvaried until the end of the treatment. These data suggest that Aloysia polystachia extract has genotoxic and cytotoxic activity, even though further investigations are required to assess which compound of the extract could be responsible for the observed effects., Italian Journal of Anatomy and Embryology, Vol. 121, No. 1 (Supplement) 2016

BAG3 protein belongs to the family of co-chaperones involved in protein quality control and in the clearance of misfolded proteins [1]. Few studies have addressed BAG3 distribution and function in the central nervous system (CNS) and little is known about the cellular localization of BAG3 during neuronal differentiation in vitro and migration in vivo. Therefore we analysed by immunofluorescence microscopy the cellular distribution of BAG3 in the PC12 cell model treated or not with NGF and in developing and adult cortex of mice brain. Our results shows that BAG3 localizes mainly in vesicle structures of the neuritic domain during cell differentiation, while in undifferentiated cells it appears confined to the cytoplasm near the nuclear membrane. These observations were corroborated by transmission electron microscopy (TEM) which revealed that in NGF-differentiated PC12 cells, BAG3 localizes into electron-dense vesicles clustered along the axon and showing the typical aspect of the large dense core vesicles (LDCVs). Interestingly, the change of BAG3 localization during neuronal differentiation was associated only to a slight increase in the total BAG3 immunoreactivity as shown by western blot analysis. In order to provide further insights on the role of BAG3 in neuronal differentiation and migration, we also analysed BAG3 localization in mice developing and adult cerebral cortex. In mouse developing cortex, BAG3 appeared to be intensely expressed in cellular processes of migrating cells, while in adult brain a low expression was detected in neuronal cell bodies and glial cells. In conclusion, our findings suggest that the presence and differential expression of BAG3 might be required for the correct development of the nervous system as well as for the maintenance of protein homeostasis., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017

The receptor activator of nuclear factor-κB ligand (RANKL), its signaling receptor RANK, and its natural decoy receptor OPG are members of the tumour necrosis factor (TNF) and TNF receptor superfamily and are best known for their essential role in controlling osteoclastogenesis. RANKL in bone has also been shown to serve as a chemoattractant for cancer cells, thus explaining the tropism of certain cancers such as breast and prostate cancer to preferentially metastasize to bone. Here, we will discuss the important role of RANKL and its possible role in the management of bone loss in patients with breast cancer., Italian Journal of Anatomy and Embryology, Vol 119, No 1 (2014)

Questo nuovo testo di istologia, destinato alle lauree triennali è stato redatto tenero conto dell'evoluzione della disciplina e delle necessità formative delle lauree triennali sanitarie, ognuna delle quali richiede uno specifico insegnamento.

Cystic fibrosis (CF) is a genetic life-shortening condition in Caucasians. Despite being a multi-organ disease, CF is classically diagnosed by symptoms of acute/chronic respiratory disease, with persistent pulmonary infections (Amaral, 2015). CF is caused by mutations in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein. Inheritance of mutant CFTR alleles results in surface liquid depletion and defective mucociliary clearance leading to pulmonary failure. Defects in CFTR perturb the regulation of many intracellular signaling pathways including the NF-kB pathway causing excessive production of pro-inflammatory mediators. Current CF therapies are directed to delay CF lung damage by restoring CFTR function and controlling abnormal inflammation. However, only few anti-inflammatory drugs are effective for CF treatments (mainly oral corticosteroids and ibuprofen), these drugs have limited beneficial effects in presence of considerable side effects. Flavonoids have been reported as promising anti-inflammatory drugs and some of them seem to act as CFTR direct activators (Amaral, 2015). From this respect, herbal remedies or plant bioactive molecules may be of great interest. To this aim, we tested the anti-inflammatory activity of apolar extracts from the roots of three Peonia species (Paeoniaceae family), namely P. rockii P. ostii and P. lactiflora, on CFTR ΔF508/ΔF508 CuFi1 cells and normal counterpart. The effects of the extracts on intrinsic as well as TNFα-induced inflammation were evaluated by determining IL-8, IL-6 and RANTES production. Further- more, to study the direct effect of the extracts on NF-kB activation, Human Embryonic Kidney cells were used in transient transfection of NF-kB reporter plasmid and NF-kB activity and cytokine productions were also evaluated. Results showed a significant anti-inflammatory activity of all three Peonia extracts with the P. lactiflora being the most effective. Furthermore, we also tested the anti-inflammatory potential of the pure flavonoid naringin in the same model systems. We found that naringin was able to reduce cytokine release through inhibiting the key enzymes of the NF-kB and MAPK/ ERK pathways. Interestingly, preliminary results on spray dried pharmaceutical formulations of this molecule, show that naringin co-sprayed with leucine improves pharmacological activity of the flavonoid neat raw drug., Italian Journal of Anatomy and Embryology, Vol. 120, No. 1 (Supplement) 2015

Osteoclasts, as well as preosteoclasts, show different adhesion features in relationship to the substrate on which cells are grown, i.e. the formation of either podosomes belt or sealing zones. Podosomes belt forms on non bone substrates, i.e. when cells interact with glass coverslips or culture plates, whereas sealing zones form when cells grow on bone-like substrates. Podosomes belt corresponds to numerous F-actin columns arranged at the cell periphery, whereas the sealing zone could be defined as a unique large band of actin [1]. In the study of bone resorption mechanisms, the employ of bone slices is not perfectly suitable to investigate actin rearrangement due to cell-extracellular matrix (ECM) interaction, since it doesn’t allow to obtain high quality preparations to be examined both by light and electronmicroscopy (TEM and SEM). In particular, TEM preparation requires demineralization which could influence the chemical properties of either bone slices or bone-like composites. Moreover, the use of bone slices as scaffold, although extensive, doesn’t allow ultrastructural details that are necessary in the study of mineral resorption by monocytes or preosteoclasts [2,3]. The aim of the present study was to set up an experimental model for the study of cell-ECM interaction between either monocytes or early differentiating osteoclasts and a mineralized ECM. RAW 264.7 cells (a monocyte-macrophage cell line that can differentiate in osteoclasts) were cultured on a composite constituted by calcium phosphate and type I collagen to investigate actin polymerization and podosome formation. This bone-like composite doesn’t present the mechanical bone properties, but it is constituted by the main bone components and exhibits the advantage that collagen glues the mineral phase in clusters that can be either added to cell cultures or applied on coverslips, as well as to the culture medium. Light and fluorescence microscopy, as well as TEM and SEM techniques were employed. Results showed that the use of this bone-like composite allowed to obtain useful morphological information about the resorption activity of RAW 264.7 cell line differentiating towards the osteoclastic phenotype., Italian Journal of Anatomy and Embryology, Vol 117, No 2 (Supplement) 2012

Mitophagy is a novel identified autophagic process through which defective and malfunctioning mitochondria are degraded or recycled. The balance between this process and mitochondrial biogenesis is crucial for post-mitotic neurons’ survival, as mitochondria are the primary energetic source required for the formation and function of axon terminals and dendritic spines (Amadoro et al 2014). As other cell types, neuronal populations also continually modulate size and number of these organelles, according to the variable energy demands and metabolic states throughout the entire lifetime and/or different sub-cellular compartments. In polarized neurons removal of damaged mitochondria can be a bioenergetically demanding task because these organelles need to be actively transported from the axons and dendrites in order to fuse to cell body resident lysosomes (Wang et al 2006). Physiologically aging as well as numerous human neurodegenerative disease, including Parkison’s disease, Huntington’s disease, amyotrophic lateral sclerosis and Alzheimer’s disease (AD) has been associated with alterations in quality control of neuronal mitochondria (Santos et al 2010), but still now, mitophagy pathway and its dysfunction in neudegenerative disease remain undisclosed. Data from several neurotoxic models show that mitochondria undergo to selective morphological changes in distribution, shape and interconnectivity which are hallmarks of mitophagy. These changes are typically studied at the confocal and TEM microscopes and quantified through a morphometric approach. Due to their polarized nature, neurons present a loss in the neuritic domain more pronounced in respect to the cell body compartment. In addition, morphometric analysis show that the mitochondria interconnectivity, area/perimeter ratio and elongation, major/minor axis ratio, are reduced in neurotoxic models and that these changes parallel in vivo findings from tissue derived from several neurodegenerative diseases. Thus, neurotoxic models, which are acute insults, are able to trigger mitophagy in similar way to human neurodegenerative diseases which, instead, develop the pathology in the course of several years., Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

Presentazione istologico funzionale del tessuto osseo

Terapia oncologica e psoriasi

Osteointegrated titanium dental implants are widely used biomaterials that have to integrate within the alveolar bone and interact with periodontal soft tissues. In this study, we investigated the immunocytochemical expression of the extra-cellular matrix (ECM) protein fibronectin (FN) and type I collagen (Coll I) in Flow 2002 fibroblast cultures spread on grade III-titanium samples with five different surface topographies and we correlated the immunocytochemical data to the adhesion capability of these cells to the above-mentioned substrates. Five different surfaces of grade III-titanium implants were at first characterized both by scanning electron microscopy (SEM) and by laser profilometry for surface roughness evaluation. After being spread on the biomaterial surfaces, the fibroblasts were left to proliferate for 72 hr and subsequently the cells underwent immunocytochemical procedures for detecting both FN and Coll I. The fibroblasts appeared more adherent to smoother titanium surfaces than to rougher ones; however, the highest cell density was detected on the roughest surface, even if it was unrelated to the highest FN expression. In the other biomaterial surfaces examined, as well as in controls, immunocytochemical FN expression correlated effectively to cell density on the examined substratum, whereas no determinant information was available regarding Coll I. It is reasonable to assume that surface roughness could be a relevant parameter influencing fibroblast adhesion to substrata; however, the evaluation of the cell density only is insufficient, and should be supported by the immunocytochemical FN expression, which could be confirmed as a useful tool in determining implant material biocompatibility. (Journal of Applied BiomaterialsBiomechanics 2004; 2: 169-76).

Bone is a complex tissue that have many function, when bone remodeling becomes perturbed we can assist at many pathologies. The key factor in these pathologies is a perturbation of RANKL/RANK/OPG pathways. We have tune up a 3D culture system that have the advantage to produce osteoclast cells more physiological than the administration of the recombinant RANKL protein. This can be useful when study new drugs to avoid the cross-reaction with the exogenous cytokines.

The role of RANK/RANKL/OPG in bone modeling