DIAGNOSIS AND PROGNOSIS OF ADRENAL BENIGN AND MALIGNANT TUMORS: NEW INSIGHTS FROM A COMPUTERIZED OPERATOR-INDEPENDENT MORPHOMETRIC MODEL AND PROTEOMIC EXPRESSION

Background: High mitotic index, high nuclear grade and reticulin distruption are part of representative hallmarks of adrenocortical cancer (ACC). A characteristic neutrophil/T-lymphocytes infiltrate ratio has been often implicated in carcinogenesis, progression and clinical outcome of several cancer types. However, its role in adrenal cortical tumors is unclear. Histology-based diagnosis may also suffer of a moment of subjectivity due to inter- and intra-observer variations. Proteomic studies of malignant tumors represent the future both for possible diagnostic and prognostic implication; whole proteome analysis of adrenocortical tumors from fresh tissues may represent the lacking piece of puzzling management of this tumor. Aim: to assess by computerized morphometry morphological features, vascular, inflammatory, reticulin and proliferative index pattern in adrenocortical adenomas (ACAs) and carcinomas and to assess proteomic profiles from adrenocortical tumors fresh tissues. Methods: A single Institution series of 11 ACAs and 18 ACCs samples was analyzed using a Kontron-Zeiss KS400 image analyzer. Four consecutive sections 4 \ub5m thick were obtained with a total of 250\u2013300 HPF examined for each case. Immunohistochemistry for Ki67, reticulin and CD8/CD15 was obtained. To minimize subjectivity, particularly relevant when quantitative results are expected, we generated a morphometric model based on analysis of volume fractions occupied by Ki67 positive and negative cells (nuclei, cytoplasm) and inflammatory compartments (CD15+ granulocytes, CD8+ lymphocytes) and reticulin framework surface. Lastly, the assessment of Ki-67 by computerized morphometry was compared with pathologist\u2019s evaluation. After sample preparation protocol of 7 ACCs, 5 ACAs and 5 normal adrenal tissue samples, difference In Gel Electrophoresis (DIGE) and following the protein spots individuation and isolation, Mass Spectrometry, were performed to allow protein identification. Results: The volume fraction of Ki-67 positive cells was highest in ACC. The volume fraction of nuclei in unit volume and the nuclear/cytoplasmic ratio in both Ki-67 negative cells and Ki-67 positive cells were prominent in ACC. The surface fraction of reticulin was considerably lower in ACC. Moreover, when comparing morphometric analysis of Ki67+ cells to pathologist\u2019s scores, the data of the point grid analysis revealed significantly lower values compared to conventional histopathology. These values, once statistically analyzed, demonstrated that our morphometric model could improve the sensitivity and specificity of Ki-67 evaluation in ACCs and ACAs (reaching 94% of sensitivity, 100% of specificity) and also that it cold contribute to a better prognosis definition. Proteomics individuated 62% overexpressed proteins in ACCs with respect to ACAs: among them vimentin and vitamin D-binding protein resulted the most varied (3.2 and 3-fold more expressed in ACCs than in ACAs respectively). On the other hand the remaining 38% of proteins resulted under-expressed in ACCs with regard to ACAs, being cathepsin D and aldose reductase both 3-fold less expressed in ACCs than in ACAs. The protein profile of ACCs versus normal adrenal tissue was similar (although with slight differences in terms of fold variation) to that of ACCs versus ACAs; nonetheless a varied new protein (lactate dehydrogenase, 1.8 fold increase in ACCs) with a possible role in tumorigenesis and tumor progression, was detected. Conclusions: Our computerized morphometric model is simple, lacks observer or subjective bias and can be used to supplement objective methods to achieve precise and reader-independent quantification of morphological characteristics and histological biomarkers of adrenocortical tumors. We speculate that the assessment of inflammatory infiltrate may found a place in the diagnostic algorithm of adrenal benign and malignant tumors. The promising preliminary results obtained by the proteomic study of ACCs and ACAs could contribute to the identification of new histological biomarkers. These data, once integrated into a complex algorithm including histological assessment, morphometric analysis and clinical data evaluation would easily contribute to create a prognostic stratification of ACCs with clear advantages for the clinical management of the disease

Who talks about collaborative spaces, how, and why

Communities in urban contexts and firms in corporate offices have recently started to implement collaborative spaces. Several authors from different disciplines are currently advancing knowledge in this realm. Systematizing this diverse knowledge base helps to advance our understanding of this novel phenomenon. To this end, the present work reviews 29 papers focusing on collaborative spaces. We analyse these papers in terms of contents, research methods, fields of study, authors’ background, and impact on the academic community. Grounding on this analysis, we outline new relevant research questions and opportunities for future investigations

1-Gb/s Transmission Over a Phosphorescent White LED by Using Rate-Adaptive Discrete Multitone Modulation

Light-emitting diodes (LEDs), which will be increasingly used in lighting technology, will also allow for distribution of broadband optical wireless signals. Visible-light communication (VLC) using white LEDs offers several advantages over the RF-based wireless systems, i.e., license-free spectrum, low power consumption, and higher privacy. Mostly, optical wireless can provide much higher data rates. In this paper, we demonstrate a VLC system based on a white LED for indoor broadband wireless access. After investigating the nonlinear effects of the LED and the power amplifier, a data rate of 1 Gb/s has been achieved at the standard illuminance level, by using an optimized discrete multitone modulation technique and adaptive bit- and power-loading algorithms. The bit-error ratio of the received data was 1.5 10^(-3), which is within the limit of common forward error correction (FEC) coding. These results twice the highest capacity that had been previously obtained

Structure and Properties of DNA Molecules Over The Full Range of Biologically Relevant Supercoiling States

Topology affects physical and biological properties of DNA and impacts fundamental cellular processes, such as gene expression, genome replication, chromosome structure and segregation. In all organisms DNA topology is carefully modulated and the supercoiling degree of defined genome regions may change according to physiological and environmental conditions. Elucidation of structural properties of DNA molecules with different topology may thus help to better understand genome functions. Whereas a number of structural studies have been published on highly negatively supercoiled DNA molecules, only preliminary observations of highly positively supercoiled are available, and a description of DNA structural properties over the full range of supercoiling degree is lacking. Atomic Force Microscopy (AFM) is a powerful tool to study DNA structure at single molecule level. We here report a comprehensive analysis by AFM of DNA plasmid molecules with defined supercoiling degree, covering the full spectrum of biologically relevant topologies, under different observation conditions. Our data, supported by statistical and biochemical analyses, revealed striking differences in the behavior of positive and negative plasmid molecules

Aquaporin 1 (Aqp1) expression in healthy dog tears

Aquaporins (AQPs) are a family of thirteen membrane proteins that play an essential role in the transport of fluids across the cell plasma membrane. Recently, the expression of AQPs in dierent ocular tissues and their involvement in the pathophysiology of eye diseases, have garnered attention. Considering that literature on AQP expression in the lacrimal glands and their secretion is scarce, we aimed to characterise AQP1 expression in the tears of healthy dogs using two tear collection methods (Schirmer tear strips (STS) and ophthalmic sponges (OS)). Fifteen healthy dogs, free of ophthalmic diseases, were included in the study. Tear collection was performed by using STS in one eye and OS in the other. After the extraction of proteins from the tears, the expression of AQP1 was analysed by Western blotting. AQP1 was expressed as a band of 28 kDa. In addition, dierences were observed in the expression of AQP1 and in the correlation between tear volume and protein concentration, in tears collected by the two dierent methods. Our results suggest that AQP1 has a specific role in tear secretion; further research is required to assess its particular role in the function of the ocular surface in eye physiology and pathology

A LOW-COST ICT SOLUTION TO SUPPORT VISITORS IN TOURISTIC CAVES

This study aims to examine innovative solutions to enhance the tourist experience and visitor localization within the Bossea Cave, one of the most important karstic caves in Italy, located in the municipality of Frabosa Soprana. The lack of advanced technological tools for managing cave structures and modernizing visitor tours creates a significant opportunity for the integration of new technologies. The researchers propose a low-cost and modular hardware infrastructure, consisting of a series of single-board computers distributed throughout the cave, acting as local web servers to provide visitors with customizable multimedia content, accessible through a web application on their personal devices via a local Wi-Fi network. This infrastructure also enables visitor localization based on their connection point within the cave, with the additional goal of testing the ultra-wideband (UWB) wireless technology in this complex and humid environment. The UWB technology offers high-precision localization, even in indoor environments and caves, where GNSS signals are not available. Overall, the study provides a promising solution to enhance the visitor experience and offers opportunities for cave management and research

The use of different adhesive filling material and mass combinations to restore class II cavities under loading and shrinkage effects: a 3D-FEA

3D tooth models were virtually restored: flowable composite resin + bulk-fill composite (A), glass ionomer cement + bulk-fill composite (B) or adhesive + bulk-fill composite (C). Polymerization shrinkage and masticatory loads were simulated. All models exhibited the highest stress concentration at the enamel–restoration interfaces. A and C showed similar pattern with lower magnitude in A in comparison to C. B showed lower stress in dentine and C the highest cusps displacement. The use of glass ionomer cement or flowable composite resin in combination with a bulk-fill composite improved the biomechanical behavior of deep class II MO cavities

The use of different adhesive filling material and mass combinations to restore class II cavities under loading and shrinkage effects: a 3D-FEA

3D tooth models were virtually restored: flowable composite resin + bulk-fill composite (A), glass ionomer cement + bulk-fill composite (B) or adhesive + bulk-fill composite (C). Polymerization shrinkage and masticatory loads were simulated. All models exhibited the highest stress concentration at the enamel-restoration interfaces. A and C showed similar pattern with lower magnitude in A in comparison to C. B showed lower stress in dentine and C the highest cusps displacement. The use of glass ionomer cement or flowable composite resin in combination with a bulk-fill composite improved the biomechanical behavior of deep class II MO cavities