Melatonin limits adaptive ER stress and hepatosteatosis in leptin-deficient mice

Non alcoholic fatty liver disease (NAFLD) impacts on about 30% of the population in industrialized countries, associated to the metabolic syndrome may be reversible or dramatically evolve into cirrhosis or hepatocellular cancer (Wree et al., 2011). Leptin-deficient homozygous mice (ob/ob) represent a well-known animal model to study obesity, associated with overweight, liver steatosis and insulinresistance. Recently ER stress has been reported to contribute to hepatic steatosis and cell damage called lipoapoptosis (Flamment et al., 2010). Melatonin, the main pineal indoleamine, has been demonstrated to be useful to limit adipogenesis in many metabolic clinical conditions (de Luxan-Delgado et al., 2014). Therefore major aims of the present study were: 1.To localize ER stress, energy homeostasis and hypoxia markers in the liver of ob/ob mice receiving or not melatonin in drinking water at 100 mg/ kg/day for 8 weeks; 2.To characterize hepatic steatosis and quantify macrosteatosis in different experimental groups. C57BL6 mice treated or not with melatonin were used as controls. Remarkably in ob/ob mice receiving melatonin, macrosteatosis, periportal GRP78 staining decreased while beta catenin became basolateral into hepatocytes. Furthermore melatonin limited nuclear CHOP staining, a recognized index of major sensitivity to apoptosis, but stimulated p62/SQSTM1 signal, involved in reducing lipogenesis. Moreover by TEM analysis, we visualized in ob/ob mice liver mitochondria that displayed more cristae and strict RER adhesion after melatonin intake. In conclusion, our morphological analysis suggests that melatonin might ameliorate NAFLD by anti-oxidative and ER stress modulatory abilities in obese mice

NPP1 inhibits intimal hyperplasia in ApoE knockout mice

Atherosclerosis is an important cause of morbidity and mortality, which is increasingly recognized and reported on a global scale. This pathology is due to multiple metabolic toxicity including increased levels of reactive oxygen species (ROS). Excessive ROS are damaging to proteins, lipids, carbohydrates and nucleic acids, which prompt a classic “response to injury” mechanism including inflammation supporting a cytokine surge, granulation and fibrosis. ROS are excessive, robustly produced in atherosclerosis associated with endothelial dysfunction. Excessive ROS due to osteopontin (OPN) increase may be the driving force promoting atherosclerotic process. Recently has been shown that ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) promotes atherosclerosis, potentially mediated by OPN expression in ApoE knockout mice (Nitschke et al., 2011). Hence, this study tested the hypothesis that NPP1 deficiency modulates intimal hyperplasia and oxidative stress in the atherosclerotic process. For this study were used ApoE null mice and Npp1/ApoE double deficient mice. Atherosclerotic lesion area, calcification and vascular alterations were examined at 13, 18, 23 and 28 weeks of age. Morphological changes in vessels were evaluated by histological procedures and immunohistochemical analysis using thrombospondin-1 (TSP-1), transforming growth factor-β1 (TGF-β1), plasminogen activator inhibitor-1 (PAI-1) and oxidative stress markers such as superoxidodismutase (SOD) and inducible nitric oxide synthase (iNOS). In ApoE null mice vessels we demonstrated vascular alterations with extensive accumulation of collagen and elastic fibers and also an increase of TSP-1, TGF-β1, PAI-1 expression and oxidative stress related protein levels compared to Npp1/ApoE double deficient mice. Moreover, histological analysis showed neointima formation only in ApoE deficient mice. Our findings suggest that NPP1 could be involved in intimal hyperplasia and oxidative stress in the atherosclerosis pathway

Symmetrical anatomical variation of the anterior belly of the digastric muscle

The digastric muscle is an important surgical landmark. Several anatomical varia- tions of the digastric muscle are reported in literature and the presence of accessory anterior bellies of the muscle are not uncommon (1,2). We reported a symmetrical variation of the digastric muscle that was found during a dissection of the suprahy- oid region. The dissection showed digastric muscles with an accessory anterior belly, which originated from the anterior belly of muscles in proximity and anterior to the intermediate tendon. The accessory bellies of both sides were fused together on the midline and were attached with a unique tendon to the inner surface of the mental symphysis, filling the submental triangle completely. This unreported anatomical var- iation could be considered an additional contribute in the description of the varia- tions of the digastric muscle, with several implications in head and neck pathology, diagnosis and surgery.

Sirtuin 6 localization at cortical brain level of young diabetic mice

The metabolic syndrome, characterized by visceral obesity, dyslipidaemia, hyperglycaemia and hypertension, has become one of the major public-health challenges worldwide and it is strictly associated with the development of type II diabetes and neurodegenerative diseases (Alberti et al. 2005; Panza et al. 2010). Increased metabolic flux to the brain during overnutrition can orchestrate stress response, blood-brain barrier alteration, microglial cells activation and neuroinflammation (Nerurkar et al., 2011). The protein sirtuin family is a class of nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase that act on a variety of targets and so play a key role in central physiological regulation (Sebastian et al., 2012; Wang et al., 2012). To assess the physiopathological significance of sirtuin6 (SIRT6) at brain cortical level, we analysed its specific expression and subcellular localization in young db/db mice, animal model of type II diabetes mellitus, and respective control lean mice. In particular, we analysed the cytoarchitecture of the brain cortex, evaluated SIRT6 expression and its localization by immunohistochemistry comparing young db/db mice to lean control mice, distinguishing among the six cortical layers and between motor and somatosensory cortex. We observed that SIRT6 is mainly localized in the nucleus of both lean and db/db mice. Diabetic mice showed few SIRT6 positive cells respect to lean control mice in all cortical layers without significant differences between motor and somatosensory cortex. No morphological alteration have been find. In conclusion, our findings contribute to further understand SIRT6 protein expression in the early steps of type II diabetes mellitus and suggest its implication in the pathogenic processes of diabetes mellitus and diabetes–induced neurodegeneration

Sirtuin 6 localization at cortical brain level of young diabetic mice

The metabolic syndrome, characterized by visceral obesity, dyslipidaemia, hyperglycaemia and hypertension, has become one of the major public-health challenges worldwide and it is strictly associated with the development of type II diabetes and neurodegenerative diseases (Alberti et al. 2005; Panza et al. 2010). Increased metabolic flux to the brain during overnutrition can orchestrate stress response, blood-brain barrier alteration, microglial cells activation and neuroinflammation (Nerurkar et al., 2011). The protein sirtuin family is a class of nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase that act on a variety of targets and so play a key role in central physiological regulation (Sebastian et al., 2012; Wang et al., 2012). To assess the physiopathological significance of sirtuin6 (SIRT6) at brain cortical level, we analysed its specific expression and subcellular localization in young db/db mice, animal model of type II diabetes mellitus, and respective control lean mice. In particular, we analysed the cytoarchitecture of the brain cortex, evaluated SIRT6 expression and its localization by immunohistochemistry comparing young db/db mice to lean control mice, distinguishing among the six cortical layers and between motor and somatosensory cortex. We observed that SIRT6 is mainly localized in the nucleus of both lean and db/db mice. Diabetic mice showed few SIRT6 positive cells respect to lean control mice in all cortical layers without significant differences between motor and somatosensory cortex. No morphological alteration have been find. In conclusion, our findings contribute to further understand SIRT6 protein expression in the early steps of type II diabetes mellitus and suggest its implication in the pathogenic processes of diabetes mellitus and diabetes–induced neurodegeneration

Platelet preparations in neuronal cell differentiation

Concentrated Growth Factors (CGF) is a platelet rich preparation that has the important feature of a tight fibrin network and containing a large number of growth factors possessing great regenerative potentialities [1]. The regeneration of nervous system is one of the mail goal of regenerative medicine. The aim of this study is to test the in vitro CGF effects on both differentiated and undifferentiated SH-SY5Y cells, derived from human neuroblastoma. To induce differentiation, SH-SY5Y cells have been treated with Retinoic Acid (RA) 10µM, in both basal and complete medium and in the presence and absence of CGF. After 72 hours, different parameters have been investigated: the morphological characteristics of the cells, the cell proliferation, the cellular vitality using the MTT test, the CGF and/or RA differentiation property and the immunocytochemical analysis of neuronal specific markers (NeuN, Sinaptophisine, β-III-tubulin, Nestin). Moreover the NGF (Nerve Growth Factor) and BDNF (Brain Derived Growth Factor) release have been assayed by ELISA test. Our results obtained suggest that treatment with CGF, also used alone, positively affects cell differentiation and neuronal phenotype regulating the expression of the neuronal markers and improving the outgrowth of neurites. Taken together these results seems to be promised into new approaches for neuronal regeneration using platelet preparations

The anastomotic network around the anterior superior alveolar nerve: an anatomical and radiological study

Innervation of superior teeth is supplied by the posterior (PSAN), anterior (ASAN) and sometimes by middle superior alveolar nerve (MSAN). PSAN arises from the maxillary nerve and passes through the posterolateral maxillary wall towards the posterior teeth. ASAN arises from the anterior portion of the infraorbital nerve and courses within the infraorbital canal passing nearby the piriform aperture and premaxilla. When present, MSAN arises from the posterior portion of the infraorbital nerve and runs along the lateral maxillary wall. However, an additional nasopalatine or sublabial injection is frequently required to obtain a complete anesthesia of the maxillary teeth due to rich anastomotic network (1-2). With the aim to better describe the complexity of the superior alveolar nerve network, fifty-seven high-definition sinonasal cone-beam CT (CBCT) were analyzed. PSAN, ASAN and MSAN were detected by specific bony landmarks/canals and nervous anastomoses were accurately evaluated. In addition, medial anastomotic branches from the palatal and/or nasal nervous plexi were also considered. PSAN and ASAN were identified in 100% of cases whereas MSAN in 19.6% of cases. Anastomotic branch versus ASAN was identified in all cases from MSAN and in 60.3% from PSAN. Medial anastomotic branch was detected in 62.0% of cases from the nasal plexus and in 6.2% from the palatal plexus: the former passed through a bony defect in the floor of the piriform aperture or at the base of the nasal septum; the latter passed through a tiny canal in the interface between maxilla and premaxilla. These data confirm that maxillary teeth innervation, especially for incisor teeth, could be provided not only by alveolar nerves but also from palatal and nasal plexi via small branches running within maxillary bony canals. These results support the need of additional anesthetic injection to obtain adequate anesthesia of the maxillary teeth; moreover, the role of CBCT in the identification of the nervous pattern was underlined

Symmetrical anatomical variation of the anterior belly of the digastric muscle

The digastric muscle is an important surgical landmark. Several anatomical varia- tions of the digastric muscle are reported in literature and the presence of accessory anterior bellies of the muscle are not uncommon (1,2). We reported a symmetrical variation of the digastric muscle that was found during a dissection of the suprahy- oid region. The dissection showed digastric muscles with an accessory anterior belly, which originated from the anterior belly of muscles in proximity and anterior to the intermediate tendon. The accessory bellies of both sides were fused together on the midline and were attached with a unique tendon to the inner surface of the mental symphysis, filling the submental triangle completely. This unreported anatomical var- iation could be considered an additional contribute in the description of the varia- tions of the digastric muscle, with several implications in head and neck pathology, diagnosis and surgery