Neurodegeneration associated-proteins in human olfactory epithelium: immunocytochemical and biomolecular study in healthy subjects and patients with synucleinopathies

Olfactory impairment is considered an initial disturbance of several neurodegenerative diseases (NDs), including Parkinson\u2019s disease (PD) and Alzheimer\u2019s disease (AD). In addition, smell impairment precedes a decade, or even longer, the onset of motor or cognitive symptoms. Olfactory signals are detected by olfactory receptor proteins (ORPs) expressed in the cilia of olfactory receptor neurons (ONs). ONs are the distinctive cellular components of the peripheral olfactory epithelium (OE) and lie in the nasal vault. ONs axons pass the cribriform plate and reach the olfactory bulb (OB) where the olfactory stimuli are processed and sent to the superior nuclei of the CNS. Previous studies in AD and other neurodegenerative disorders have shown the presence of \u3b2-amyloid deposits in the OB, neurofibrillary tangles, as well as Lewy body pathology. OB represents the brain area earlier involved in the neuropathological process, decades before the development of clinical symptoms. Therefore, OB can be considered a target in the study of neurodegenerative diseases in their early molecular processes. Moreover, the OB of healthy subjects presents deposits of aggregated proteins confirming that these aggregates are deposited in a prodromal disease stage. Since the OB is an early accumulation site of aggregated proteins and the synapses derive from the ONs, it is possible that the first event of protein aggregation occurs in OE. ONs are directly exposed to the external environment including chemical/physical toxic injuries and such micro-environment predisposes to abnormal protein processing and folding (Sammeta and McClintock 2010). In addition, ONs and all other mature cell components have a half-life of three months and programmed apoptosis. The neural activity is maintained by a constant cellular turn-over, which is sustained by the basal stem cells. This regeneration process is persistent during the whole life of an individual, albeit with a decreasing rate with aging. Extensive scientific literature indicates the neuronal damage as the consequence of exposure to toxic injuries leading to neurodegeneration and ONs are a natural model of this noxious process (Lema Tom\ue9, Tyson et al. 2013). The hypothesis of this pathological pathway is supported by several studies, in which aggregated forms of \u3b1-synuclein, tau and \u3b2-amyloid are detected in olfactory mucosa (OM) biopsies as well as in autoptic samples of patients with Parkinson\u2019s disease (PD), Lewy body dementia (LBD), Frontotemporal dementia (FTD) and Alzheimer disease (AD) (Funabe, Takao et al. 2013) (Saito, Shioya et al. 2016) (Tabaton, Cammarata et al. 1991) (Talamo, Rudel et al. 1989) (Crino, Greenberg et al. 1995) (Arnold, Lee et al. 2010). In this study, we investigated for the first-time primary ONs sampled ex vivo using olfactory brushing (OBg) in normal subjects and patients with different neurodegenerative disorders. Because of its convenient location, OE is easily accessible and can be sampled to obtain the ONs in the tissue outer layer. This sampling method is harmless and non-invasive, bypassing potential artifacts due to post mortem specimens as well as avoiding the invasiveness of biopsy procedures. Recently, we showed that OBg procedure in Creutzfeldt-Jakob Disease (CJD) patients allows efficient OM sampling for the Real-Time Quaking-Induced Conversion (RT-QuIC) assay. We specifically amplified pathological prion protein (PrPSc) providing a diagnostic intra vitam test with sensitivity and specificity nearly to 100% (Orr\ufa, Bongianni et al. 2014). For the purpose of our study, we firstly defined the phenotypic characterization of the human olfactory cells sampled with OBg from healthy subjects. Distinct antibodies were selected to analyze the olfactory epithelium cells: olfactory marker protein (OMP), neuron-specific class III \u3b2-tubulin (TUJ-1), protein gene product 9.5 (PGP 9.5), Pan-Cytokeratin (PCK). Secondly, we aimed to determine the expression patterns of the major misfolded proteins involved in the main neurodegenerative diseases. In particular, the selected proteins were: \u3b1-synuclein, APP/beta-amyloid, tau, and TDP-43. The identification of the expression patterns of these proteins in the ONs might provide information to understand the abnormal molecular mechanisms in the initial misfolding species involved in the pathological process. Moreover, in this study, we speculated on the subcellular locale where the protein aggregation may occur. Furthermore, by demonstrating the constitutive expression of the native NDs-associated proteins in the OE, we could assume that they may represent a potential template for triggering the aggregation process. Based on the immunocytochemistry analysis, we investigated the \u3b1-synuclein expression in patients affected by different synucleinopathies. In fact, \u3b1-synuclein misfolding and aggregation mechanisms are involved in the pathogenesis of neurodegenerative disorders such as Parkinson's disease (PD), dementia with Lewy bodies (LBD) and multiple system atrophy (MSA), which are all characterized by \u3b1-synuclein fibrils deposition (Spillantini, Schmidt et al. 1997). Finally, we analyzed the immunocytochemistry results in OM samples tested by \u3b1-synuclein RT-QuIC (\u3b1-syn RT-QuIC)

Welding parameters and their effect of column strength

This thesis presents a study on the influence of different welding parameters on the magnitude and distribution of residual stresses in flame-cut plates 24\u27 x 2\u27 made of A36 steel. The residual stress diagrams obtained after complete sectioning and after slicing are related to the original conditions of fabrication and mnaufacturing

Robot-assisted pancreaticoduodenectomy. Safety and feasibility

Background: The availability of robotic assistance could make laparoscopic pancreaticoduo- denectomy safely feasible. We herein provide a systematic review on laparoscopic robot-assisted pancreaticoduodenectomy (RAPD). Methods: Literature search was conducted on multiple databases considering articles published in English up to October 31, 2014, reporting on ten or more patients. Results: A total of 262 articles were identified. Excluding duplicates (n=172), studies not matching inclusion criteria (n=77), and studies not suitable for other reasons (n=6), a total of seven studies reporting on 312 RAPDs were eventually reviewed. These studies were either retrospective cohort studies (n=4) or case-matched studies (n=3). No randomized controlled trial was identified. Most patients undergoing RAPD were diagnosed with malignant tumors (224/312; 71.8%). RAPD was feasible in most patients. Conversion to open surgery was reported in 9.2% of the patients. A hybrid RAPD technique, employing standard laparoscopy or open surgery through a mini-incision, was adopted in most patients (178/312; 57.0%). Overall, there were six postoperative deaths at 30 days (6/312; 1.9%), including one intraoperative death caused by portal vein injury, while 137 out of 260 patients with complete information developed postoperative complications (52.7%). The mean length of hospital stay ranged from 10–29 days. Postoperative pancreatic fistula (POPF) occurred in 66 patients (66/312; 21.1%). Grade C POPF was reported in eight patients (8/312; 2.5%). The costs of RAPD were assessed in two studies, demonstrating additional costs ranging from 4,000–5,000 US dollars to 6,193 Euro. The mean number of examined lymph nodes and the rate of positive surgical margins indicate that RAPD could be an appropriate oncologic operation. Conclusion: RAPD is safely feasible. These results were obtained in selected patients and in specialized centers. RAPD should not be implemented in the occasional patient by surgeons without advanced laparoscopic skills and formal training in robotic surgery

Systematic review and metanalysis of clinical outcomes after enucleation of pancreatic metastases from renal cell carcinoma

Introduction: A systematic review of the literature was carried out to determine the clinical and oncological outcome of patients who had enucleation of solitary pancreatic metastases from renal cell carcinoma. Methods: Operative mortality, postoperative complications, observed survival and disease free survival were analyzed. The clinical outcomes of patients who had enucleation were compared to those of 857 patients collected from the literature who had standard or atypical pancreatic resection for the same disease using propensity score matching Results: There was no postoperative mortality in the 56 patients who had enucleation of pancreatic metastases from renal cell carcinoma . In 51 patients postoperative complications could be analyzed. Ten patients (10/51=19.6%) had postoperative complications. Three patients (3/51 = 5.9%) had major complications (Clavien-Dindo III or more). Five year observed survival rates and disease free survival for patients with enucleation were 92% and 79% respectively. These results compared favourably with those obtained in patients who had standard resection and other forms of atypical resection (also using propensity score matching). Patients who had partial pancreatic resection (atypical or not) with pancreatic-jejunal anastomosis had increased rates of postoperative complications and local recurrences. Conclusions: Enucleation of pancreatic metastases offers a valid solution in selected patients

La relazione tra intermediari e clienti: un rapido sguardo d’assieme sui profili evolutivi

Il saggio ripercorre in rapida sintesi le tappe salienti della regolazione del rapporto tra intermediari e clienti, con un richiamo delle finalità della stessa, degli aspetti problematici, delle ricadute sui profili organizzativi degli intermediari bancari. Vengono sottolineate la “fluidità” del complessivo apparato regolatorio, sempre più pervasivo e con struttura “multistrato”, nonché la difficoltà del diritto dell’economia di afferrare un fenomeno per sua natura articolato e mutevole nel tempo.The paper provides a brief overview of the key steps in the regulatory framework about the relationship between intermediaries and customers, recalling the aims, the problematic aspects, and the effects on the organisational aspects of banking intermediaries. It highlights the "fluidity" of the regulatory system as a whole – that is ever more intrusive and “multi-layered” - and the difficulty of economic law to regulate an ever-changing phenomenon