Unsolved questions and preferred solution about living will.

Background: Ethical problems about end-of-life medicine include a variety of issues approached in different ways by physicians and, more recently, special emphasis to this kind of ethical issues and possible answers has done by Italian National Ethical Committee in the issue named “Deep and continuous palliative sedation in the imminence of Death” (January, 2016). The debate is very critical in Intensive Care Units and Cancer Wards, where health care professionals face-off with terminally-ill patients is an outright routine; the Authors investigated their medical knowledge and ethical perception about patient critical and terminal condition to discuss the most relevant conclusions. Material: In the Sicilian province of Palermo, physicians working in Intensive Care and Oncology fields were been given a questionnaire that takes inspiration from the Ethicatt Questionnaire-Doctor. The authors reported the results obtained, by selecting and analyzing the most involved questions about living wills. Results: Generally, the respondents showed a great sensibility on this topic. Overall agreement on the living will was observed, as past surveys, but also a new conception. Euthanasia remains not very popular, attitude in line with other countries. Opinions and aptitudes of relatives have minor importance towards patient’s wishes, that are in some cases in first place. Conclusion: Explicit positive answer towards dilemmas about living wills lifts the veil and reveals how these ones would represent a very useful tool for health care professionals in this study. It is also plausible that, if doctors had available an advance directive (living will) document, they would follow it, overcoming any contingent ethical objections

Time-to-digital converters and histogram builders in SPAD arrays for pulsed-LiDAR

Light Detection and Ranging (LiDAR) is a 3D imaging technique widely used in many applications such as augmented reality, automotive, machine vision, spacecraft navigation and landing. Pulsed-LiDAR is one of the most diffused LiDAR techniques which relies on the measurement of the round-trip travel time of an optical pulse back-scattered from a distant target. Besides the light source and the detector, Time-to-Digital Converters (TDCs) are fundamental components in pulsed-LiDAR systems, since they allow to measure the back-scattered photon arrival times and their performance directly impact on LiDAR system requirements (i.e., range, precision, and measurements rate). In this work, we present a review of recent TDC architectures suitable to be integrated in SPAD-based CMOS arrays and a review of data processing solutions to derive the TOF information. Furthermore, main TDC parameters and processing techniques are described and analyzed considering pulsed-LiDAR requirements

Fast-Gated 16 x 16 SPAD Array With 16 on-Chip 6 ps Time-to-Digital Converters for Non-Line-of-Sight Imaging

We present the design and characterization of a fully-integrated array of 16 x 16 Single-Photon Avalanche Diodes (SPADs) with fast-gating capabilities and 16 on-chip 6 ps time-to-digital converters, which has been embedded in a compact imaging module. Such sensor has been developed for Non-Line-Of-Sight imaging applications, which require: i) a narrow instrument response function, for a centimeter-accurate single-shot precision; ii) fast-gated SPADs, for time-filtering of directly reflected photons; iii) high photon detection probability, for acquiring faint signals undergoing multiple scattering events. Thanks to a novel multiple differential SPAD-SPAD sensing approach, SPAD detectors can be swiftly activated in less than 500 ps and the full-width at half maximum of the instrument response function is always less than 75 ps (60 ps on average). Temporal responses are consistently uniform throughout the gate window, showing just few picoseconds of time dispersion when 30 ns gate pulses are applied, while the differential non-linearity is as low as 250 fs. With a photon detection probability peak of 70% at 490 nm, a fill-factor of 9.6% and up to 1.6 . 10(8) photon time-tagging measurements per second, such sensor fulfills the demand for fully-integrated imaging solutions optimized for non-line-of-sight imaging applications, enabling to cut exposure times while also optimizing size, weight, power and cost, thus paving the way for further scaled architectures

Sexual Dimorphism in Cellular and Molecular Features in Human ACTH-Secreting Pituitary Adenomas.

Background. Cushing\u2019s disease presents gender disparities in prevalence and clinical course. Little is known, however, about sexual dimorphism at the level of the corticotrope adenoma itself. The aim of the present study was to evaluate molecular features of ACTH-secreting pituitary adenomas collected from female and male patients with Cushing\u2019s disease. (2) Methods. We analyzed 153 ACTH-secreting adenomas collected from 31 men and 122 women. Adenomas were established in culture and ACTH synthesis and secretion assessed in basal conditions as well as during incubation with CRH or dexamethasone. Concurrently, microarray analysis was performed on formalin-fixed specimens and differences in the expression profiles between specimens from male and female patients identified. (3) Results. ACTH medium concentrations in adenomas obtained from male patients were significantly lower than those observed in adenomas from female patients. This could be observed for baseline as well as modulated secretion. Analysis of corticotrope transcriptomes revealed considerable similarities with few, selected differences in functional annotations. Differentially expressed genes comprised genes with known sexual dimorphism, genes involved in tumour development and genes relevant to pituitary pathophysiology. (4) Conclusions. Our study shows for the first time that human corticotrope adenomas present sexual dimorphism and underlines the need for a gender-dependent analysis of these tumours. Differentially expressed genes may represent the basis for gender-tailored target therapy

Role of the ubiquitin/proteasome system on ACTH turnover in rat corticotropes

Purpose: A large number of studies has investigated proopiomelanocortin processing in anterior pituitary corticotropes but little is known on proopiomelanocortin/ACTH degradation within these cells. The ubiquitin-proteasome system is an intracellular protein degradation pathway which has garnered considerable interest in recent times, given its role in maintenance of protein homeostasis. Aim of the present study was to evaluate the role of the ubiquitin-proteasome system in proopiomelanocortin/ACTH turnover in pituitary corticotropes. Methods: Rat anterior pituitary primary cultures were treated with 0.01\u2013100 nM MG132, a proteasome inhibitor, or 0.1\u2013100 nM K48R, an inhibitor of polyubiquitylation, for 4 and 24 h and ACTH concentrations in medium and cell lysates estimated by immunometric assay. Co-immunoprecipitation for ubiquitin and ACTH was carried out to establish ubiquitin-tagged protein products. Results: Inhibition of proteasome-mediated degradation with MG132 lead to an increase in ACTH concentrations, both as regards secretion and cell content. Likewise, inhibition of polyubiquitylation was associated with increased ACTH secretion and cell content. Ubiquitin/ACTH co-immunoprecipitation revealed that proopiomelanocortin was a target of ubiquitylation. Conclusions: We provide the first evidence that the ubiquitin-proteasome system is involved in proopiomelanocortin/ACTH degradation in corticotropes. Indeed, proopiomelanocortin is a target of ubiquitylation and modulation of ubiquitin-proteasome system affects ACTH turnover. This study shows that regulation of ACTH proteolytic degradation may represent a means to control ACTH secretion

Ubiquitin-Specific Protease 8 Mutant Corticotrope Adenomas Present Unique Secretory and Molecular Features and Shed Light on the Role of Ubiquitylation on ACTH Processing

Background: Somatic mutations in the ubiquitin-specific protease 8 (USP8) gene have recently been shown to occur in ACTH-secreting pituitary adenomas, thus calling attention to the ubiquitin system in corticotrope adenomas. Objectives: Assess the consequences of USP8 mutations and establish the role of ubiquitin on ACTH turnover in human ACTH-secreting pituitary adenomas. Methods: USP8 mutation status was established in 126 ACTH-secreting adenomas. Differences in ACTH secretion and POMC expression from adenoma primary cultures and in microarray gene expression profiles from archival specimens were sought according to USP8 sequence. Ubiquitin/ACTH coimmunoprecipitation and incubation with MG132, a proteasome inhibitor, were performed in order to establish whether ubiquitin plays a role in POMC/ACTH degradation in corticotrope adenomas. Results: USP8 mutations were identified in 29 adenomas (23%). Adenomas presenting USP8 mutations secreted greater amounts of ACTH and expressed POMC at higher levels compared to USP wild-type specimens. USP8 mutant adenomas were also more sensitive to modulation by CRH and dexamethasone in vitro. At microarray analysis, genes associated with endosomal protein degradation and membrane components were downregulated in USP8 mutant adenomas as were AVPR1B, IL11RA, and PITX2. Inhibition of the ubiquitin-proteasome pathway increased ACTH secretion and POMC itself proved a target of ubiquitylation, independently of USP8 sequence status. Conclusions: Our study has shown that USP8 mutant ACTH-secreting adenomas present a more "typical" corticotrope phenotype and reduced expression of several genes associated with protein degradation. Further, ubiquitylation is directly involved in intracellular ACTH turnover, suggesting that the ubiquitin-proteasome system may represent a target for treatment of human ACTH-secreting adenomas

Gene expression profiling in human corticotrope tumors reveals distinct, neuroendocrine profiles

ACTH-secreting pituitary adenomas give rise to a severe endocrinological disorder, i.e., Cushing's disease, with multifaceted clinical presentation and treatment outcomes. Experimental studies suggested that disease variability is inherent to the pituitary tumor, thus pointing to the need for further studies into tumor biology. Aim of the present study was to evaluate transcriptome expression pattern in a large series of ACTH-secreting pituitary adenoma specimens, in order to identify molecular signatures of these tumors. Gene expression profiling of formalin-fixed paraffin-embedded specimens from 40 human ACTH-secreting pituitary adenomas revealed significant expression of genes involved in protein biosynthesis and ribosomal function, in keeping with neuroendocrine cell profile. Unsupervised cluster analysis identified three distinct gene profile clusters and several genes were uniquely overexpressed in a given cluster, accounting for different molecular signatures. Of note, gene expression profiles were associated with clinical features such as age and size of the tumor. Altogether, our study shows that corticotrope tumors are characterized by neuroendocrine gene expression profile and present subgroup-specific molecular features

SOLUS: An innovative multimodal imaging system to improve breast cancer diagnosis through diffuse optics and ultrasounds

To improve non-invasively the specificity in the diagnosis of breast cancer after a positive screening mammography or doubt/suspicious ultrasound examination, the SOLUS project developed a multimodal imaging system that combines: B-mode ultrasound (US) scans (to assess morphology), Color Doppler (to visualize vascularization), shear-wave elastography (to measure stiffness), and time domain multi-wavelength diffuse optical tomography (to estimate tissue composition in terms of oxy- and deoxy-hemoglobin, lipid, water, and collagen concentrations). The multimodal probe arranges 8 innovative photonic modules (optodes) around the US transducer, providing capability for optical tomographic reconstruction. For more accurate estimate of lesion composition, US-assessed morphological priors can be used to guide the optical reconstructions. Each optode comprises: i) 8 picosecond pulsed laser diodes with different wavelengths, covering a wide spectral range (635-1064 nm) for good probing of the different tissue constituents; ii) a large-area (variable, up to 8.6 mm2) fast-gated digital Silicon Photomultiplier; iii) the acquisition electronics to record the distribution of time-of-flight of the re-emitted photons. The optode is the basic element of the optical part of the system, but is also a stand-alone, ultra-compact (about 4 cm3) device for time domain multi-wavelength diffuse optics, with potential application in various fields

SOLUS: a novel multimodal approach to ultrasound and diffuse optics imaging of breast cancer

A multimodal instrument for breast imaging was developed, combining ultrasound (morphology), shear wave elastography (stiffness), and time domain multiwavelength diffuse optical tomography (blood, water, lipid, collagen) to improve the non-invasive diagnosis of breast cancer

Minimal residual disease negativity by next-generation flow cytometry is associated with improved organ response in AL amyloidosis

Light chain (AL) amyloidosis is caused by a small B-cell clone producing light chains that form amyloid deposits and cause organ dysfunction. Chemotherapy aims at suppressing the production of the toxic light chain (LC) and restore organ function. However, even complete hematologic response (CR), defined as negative serum and urine immunofixation and normalized free LC ratio, does not always translate into organ response. Next-generation flow (NGF) cytometry is used to detect minimal residual disease (MRD) in multiple myeloma. We evaluated MRD by NGF in 92 AL amyloidosis patients in CR. Fifty-four percent had persistent MRD (median 0.03% abnormal plasma cells). There were no differences in baseline clinical variables in patients with or without detectable MRD. Undetectable MRD was associated with higher rates of renal (90% vs 62%, p = 0.006) and cardiac response (95% vs 75%, p = 0.023). Hematologic progression was more frequent in MRD positive (0 vs 25% at 1 year, p = 0.001). Altogether, NGF can detect MRD in approximately half the AL amyloidosis patients in CR, and persistent MRD can explain persistent organ dysfunction. Thus, this study supports testing MRD in CR patients, especially if not accompanied by organ response. In case MRD persists, further treatment could be considered, carefully balancing residual organ damage, patient frailty, and possible toxicity