MODELLI SPERIMENTALI DI SOVRACCARICO DIETETICO DI FERRO: EFFETTI CENTRALI E PERIFERICI SU METABOLISMO E FUNZIONE RIPRODUTTIVA

Background and Aim. Iron is an essential micronutrient, which is involved as a cofactor in fundamental biochemical activities, and it is necessary for proper brain development in the fetal and early neonatal period. However, cellular iron overload produces toxic build-up in many organs, including brain, and, under aerobic conditions, catalyses the propagation of reactive oxygen species and the generation of highly reactive radicals through Fenton Chemistry. Association between metabolic and reproductive impairment has been proved in patients affected by dysmetabolic iron overload syndrome (DIOS). In particular, iron is the most important factor afflicting the hypothalamic-pituitary axis in a dose-dependent fashion leading to hypogonadotropic hypogonadism (HH). Our previous studies in a mouse model of DIOS showed the association between dietary iron overload, visceral adipose tissue insulin resistance and hypertriglyceridemia. Aim of this thesis was to assess whether and how iron overload may affect (a) the reproductive axis (mainly at the hypothalamic-pituitary levels) in a mouse model of DIOS; (b) the migratory feature and GnRH secretory pattern in GN-11 and GT1-7 cells, in vitro models of immature/migratory and mature/GnRH-secreting neurons, respectively. Results. In male mice, dietary-iron overload (IED) led to: a) an increment in testis iron content, b) a reduction in testicular weight and length, c) no changes in hypothalamic iron content c) no changes in mRNA levels of iron-responsive genes, transferrin receptor (TfR) and ferritin H (FtH), in testes and hypothalamus d) an up-regulation of hypothalamic GnRH mRNA levels, e) no changes in hypothalamic Kiss1 and GPR54 gene expression, e) a reduction in pituitary LH\u3b2 gene expression. Moreover, the hypothalamic increment of TNF\u3b1 gene expression along with the phosphorylation/activation of AMPK protein suggested the presence of an inflammatory condition. Increased hypothalamic CHOP mRNA levels also confirmed the endoplasmic reticulum stress feature. IED mice gained less weight than controls showing a reduction in VAT mass and in serum leptin levels, whereas hypothalamic NPY mRNA levels were increased and POMC gene expression was reduced. Western blot analysis showed that the pAkt/Akt ratio was up-regulated in the hypothalamus of IED mice, whereas phosphorylation of ERK1/2 (pERK) protein resulted unchanged in both groups. As far as GN-11 and GT1-7 cells are concerned, a 24-hour treatment with 200 \ub5M Ferric Ammonium Citrate (FAC, source of ferric iron) induced an increment in the intracellular specific iron content of both cell-based models without affecting the cell viability and morphology. Gene expression analysis showed that both cell lines express TfR and FtH, whose mRNA levels were modulated by iron overload. Exposure of GN-11 cells to FAC resulted in the dose (200\u20131000 \ub5M FAC for 24 hours)- and time (24-72 hours with 200 \ub5M FAC)-dependent inhibition of FBS-induced chemomigration, as assessed by Boyden chamber assay. Pre-treatment with 200 \ub5M deferoxamine (DFO, a specific iron chelator) reverted the above reported iron-driven effect on cell migration. Time-course experiments showed that 200 \u3bcM FAC was associated with increased pERK1/2 and pAkt protein levels and with decreased pAMPK ones. Chemomigration assays carried out with the specific inhibitors of ERK1/2, Akt and AMPK highlighted that only Akt pathway seems involved in FAC-mediated inhibition of GN-11 cell migration. In GN-11 cells, iron treatment increased IL-6 gene expression in a dose-dependent mode, whereas NF-kB nuclear translocation and activation was not affected. Up-regulated SOD2 mRNA levels confirmed a condition of activated oxidative stress. Conclusions. The present data show that dietary-iron overload impairs the reproductive axis, probably leading to HH, but further experiments are needed to understand the anatomic site mainly involved in iron-driven damage. Iron treatment negatively affects the migration of GN-11 neuronal cells by the activation of Akt signaling pathway. Hence, iron overload may impair the migration of GnRH neurons from the olfactory placode into forebrain and hypothalamus, where these neurons promote the reproductive competence

Recycling Aluminum

Students will investigate and compare the energy cost to produce aluminum products from aluminum ore and recycled aluminum. Students will perform an electrolysis activity to reinforce the idea that recycling metal requires less energy than mining and refining metals from their original source in the earth

The i148m Pnpla3 polymorphism influences serum adiponectin in patients with fatty liver and healthy controls

BACKGROUND: Reduced adiponectin is implicated in the pathogenesis of nonalcoholic fatty liver disease (NAFLD) and steatohepatitis (NASH), and the I148M Patatin-like phospholipase domain-containing 3 (PNPLA3) polymorphism predisposes to NAFLD and liver damage progression in NASH and chronic hepatitis C (CHC) by still undefined mechanisms, possibly involving regulation of adipose tissue function. Aim of this study was to evaluate whether the I148M PNPLA3 polymorphism influences serum adiponectin in liver diseases and healthy controls. METHODS: To this end, we considered 144 consecutive Italian patients with NAFLD, 261 with CHC, 35 severely obese subjects, and 257 healthy controls with very low probability of steatosis, all with complete clinical and genetic characterization, including adiponectin (ADIPOQ) genotype. PNPLA3 rs738409 (I148M) and ADIPOQ genotypes were evaluated by Taqman assays, serum adiponectin by ELISA. Adiponectin mRNA levels were evaluated by quantitative real-time PCR in the visceral adipose tissue (VAT) of 35 obese subjects undergoing bariatric surgery. RESULTS: Adiponectin levels were independently associated with the risk of NAFLD and with the histological severity of the disease. Adiponectin levels decreased with the number of 148\u2009M PNPLA3 alleles at risk of NASH both in patients with NAFLD (p\u2009=\u20090.03), and in healthy subjects (p\u2009=\u20090.04). At multivariate analysis, PNPLA3 148\u2009M alleles were associated with low adiponectin levels (<6\u2009mg/ml, median value) independently of NAFLD diagnosis, age, gender, BMI, and ADIPOQ genotype (OR 1.67, 95% c.i. 1.07-2.1 for each 148\u2009M allele). The p.148\u2009M PNPLA3 variant was associated with decreased adiponectin mRNA levels in the VAT of obese patients (p\u2009<\u20090.05) even in the absence of NASH. In contrast, in CHC, characterized by adiponectin resistance, low adiponectin was associated with male gender and steatosis, but not with PNPLA3 and ADIPOQ genotypes and viral features. CONCLUSIONS: The I148M PNPLA3 variant is associated with adiponectin levels in patients with NAFLD and in healthy subjects, but in the presence of adiponectin resistance not in CHC patients. The I148M PNPLA3 genotype may represent a genetic determinant of serum adiponectin levels. Modulation of serum adiponectin might be involved in mediating the susceptibility to steatosis, NASH, and hepatocellular carcinoma in carriers of the 148\u2009M PNPLA3 variant without CHC, with potential therapeutic implications

Development and implementation of a prescription opioid registry across diverse health systems

Objective: Develop and implement a prescription opioid registry in 10 diverse health systems across the US and describe trends in prescribed opioids between 2012 and 2018. Materials and Methods: Using electronic health record and claims data, we identified patients who had an outpatient fill for any prescription opioid, and/or an opioid use disorder diagnosis, between January 1, 2012 and December 31, 2018. The registry contains distributed files of prescription opioids, benzodiazepines and other select medications, opioid antagonists, clinical diagnoses, procedures, health services utilization, and health plan membership. Rates of outpatient opioid fills over the study period, standardized to health system demographic distributions, are described by age, gender, and race/ethnicity among members without cancer. Results: The registry includes 6 249 710 patients and over 40 million outpatient opioid fills. For the combined registry population, opioid fills declined from a high of 0.718 per member-year in 2013 to 0.478 in 2018, and morphine milligram equivalents (MMEs) per fill declined from 985 MMEs per fill in 2012 to 758 MMEs in 2018. MMEs per member declined from 692 MMEs per member in 2012 to 362 MMEs per member in 2018. Conclusion: This study established a population-based opioid registry across 10 diverse health systems that can be used to address questions related to opioid use. Initial analyses showed large reductions in overall opioid use per member among the combined health systems. The registry will be used in future studies to answer a broad range of other critical public health issues relating to prescription opioid use

Adiponectin interactions in bone and cartilage biology and disease

The adipokine adiponectin promotes insulin sensitivity and fat \u3b2-oxidation. In addition to its metabolic effects, adiponectin is an important local and systemic modulator of bone remodeling and cartilage biology, involving direct and indirect mechanisms and a large set of downstream molecular signals. Moreover, data suggest that changes in adiponectin signaling may be associated with bone and cartilage diseases. Adiponectin seems to exert a negative net effect on bone mass and to be an independent predictor of lower bone mass, whereas available data about actions on cartilage are more controversial, showing both pro- and anti-inflammatory actions. Adiponectin-bone cross talk seems to be reciprocal, as osteocalcin, produced by osteoblasts, has been shown to stimulate adiponectin expression and to improve glucose tolerance. Adiponectin-related signaling in bone and cartilage should be considered within the network of hormonal and nutritional signals that may influence skeleton biology, together with body homeostasis and adipose mass changes

Somatostatin (SS), SS receptors and SS analog treatment in tumorigenesis

Deep insight on Somatostatin (SS), SS receptors and SS analog treatment in tumorigenesis

Front panel human interface for FASTBUS

A human interface based on the Snoop diagnostic module has been designed to facilitate checkout of FASTBUS devices, diagnosis of system faults, and monitoring of system performance. This system, which is a generalization of the usual computer front panel or control console, includes logic analyzer functions, display and manual-control access to other modules, a microprocessor which allows the user to create and execute diagnostic programs and store them on a minifloppy disk, and a diagnostic network which allows remote console operation and coordination of information from multiple segments' Snoops

Somatostatin, Somatostatin Analogs and Somatostatin Receptor Dynamics in The Biology of Cancer Progression

The pharmacological effects (i.e., inhibition of endocrine secretion and cell proliferation) mediated by the hormone somatostatin (SRIF) are derived from its universal high-affinity binding to five different G proteincoupled receptors (GPCRs), named sst1-5. However, SRIF has a half-life of less than 3 min, whereas the available mono- and bi-specific SRIF preferential analogs show prolonged half-life and increased potency. These compounds may control tumor development, cell proliferation and metastatization by direct actions, including cell division arrest in G0/G1 phase (i.e., induction of cyclin-dependent kinase inhibitor p27(kip1) or p21(Cip1)), induction of apoptosis (i.e., induction of p53 and Bax) and suppression of cell invasion. Along with these direct actions on the biology of cancer progression, in vivo SRIF analogs may also regulate tumor growth through indirect actions, by suppressing the secretion of growth-promoting hormones and growth factors and angiogenesis. Interestingly, when ssts are co-expressed, they may interact forming homo- or heterodimers, also with other GPCRs such as type 2 dopamine receptor and the \u3bc-opioid receptor 1, altering their original pharmacological and functional properties. Dimers can be not only constitutive, but perhaps also ligandpromoted: hence, compounds with high affinity for different ssts isoforms may be used to achieve effects elicited by specific dimers. Future developments in the knowledge of ssts dynamics upon SRIF and SRIF analogs binding in neoplastic tissues may allow the full elucidation of the pathophysiological role of this system and the exploitation of the therapeutic potential of its modulation