The effects of sleep deprivation and poor sleep quality on Emotional Empathy: the behavioral and neural mechanisms in healthy controls

This dissertation describes a series of experiments exploring the intricate relationship between sleep and Emotional Empathy, i.e., the ability to understand someone else’s emotions through vicarious sharing. First, I used a sleep deprivation protocol, in which sleep quality was experimentally manipulated, to test the hypothesis that sleep deprivation negatively affects the Emotional Empathy of healthy individuals (Chapter 3). The findings of this study revealed a blunting of Emotional Empathy responses across all valences for participants in the sleep deprivation group compared to participants in the control groups. I then tested the hypothesis that sleep quality accounts for variability in individuals’ empathic responses by looking at effects of natural occurring changes in sleep quality on Emotional Empathy (Chapter 4). In this study, I collected subjective (questionnaires) and objective (actigraphy) measures of sleep and used a sophisticated statistical analysis to reduce the number of collected sleep variables and generate independent factors that were then entered into a series of stepwise regressions. The results of this study showed that the subjective sleep quality factor best explained participants’ Emotional Empathy responses to negative images compared to neutral, while the total sleep duration factor best explained overall Empathy scores. Finally, to test the hypothesis of a modulatory role of sleep quality on the neural activity of areas identified as components of the Emotional Empathy network, I conducted a Region of Interest (ROI) analysis, and measured Blood Oxygen Level Dependent (BOLD) signal change while participants performed an Emotional Empathy task. In addition, I tested the effects of sleep quality on the task based functional connectivity between the selected ROIs. The data revealed decreased BOLD signal change in a selective region within the left anterior insula for individuals with poor subjective sleep quality together with increased functional connectivity between subcomponents of the anterior insula, indicating lower functional specialization. Overall, these studies suggest a detrimental effect of poor sleep quality on Emotional Empathy and its underlying neural mechanisms. These findings could benefit individuals affected by sleep disorders but also bring insight on the importance of considering sleep loss in daily life as a detrimental factor when planning work schedules

Sleep Quality, Empathy, and Mood During the Isolation Period of the COVID-19 Pandemic in the Canadian Population: Females and Women Suffered the Most

Objective: To investigate the sex and gender differences in the impact of the isolation period implemented in response to the COVID-19 pandemic on individuals' sleep quality, empathy, and mood. Design: Data were collected between March 23 and June 7, 2020 on a sample of volunteers in the Canadian population. Six hundred and thirty-eight volunteers completed an online survey (~30 min). Main Outcome and Measures: We first examined biological sex, gender, and sexual identity differences (both components of the ampler concept of gender) in sleep, empathy, and mood disturbances. Then, we assessed changes in sleep and mood over the course of the isolation period and tested for significant relationships between sleep variables, mood, and empathy. Results: We analyzed complete data for 573 participants (112 males and 459 females, 2 undisclosed, mean ± SD age = 25.9 ± 10.5 years, mean ± SD education = 16.2 ± 2.9 years). As compared to males, female participants reported lower quality of sleep, lower sleep efficiency, and greater symptoms of insomnia, anxiety, depression, and trauma. In addition, females reported higher scores than males on the IRI empathy scale and all its subcomponents. Similar results were found when stratifying by gender. Sleep and mood disturbances increased over the course of the isolation period in the whole sample. The most significant predictors of poor quality of sleep and insomnia were depression, anxiety, and trauma scores, especially in females; higher empathy trait was associated with higher depression, anxiety, and trauma scores, perhaps indicating a more positive role of fear and anxiety responses to the pandemic crisis. Significance and Conclusions: Sex and gender differences seem to play a role in the individuals' psychological and behavioral reactions to the COVID-19 pandemic. These differences need to be considered in planning targeted psychological interventions

Poor sleep quality affects spatial orientation in virtual environments

Sleep is well known to have a significant impact on learning and memory. Specifically, studies adopting an experimentally induced sleep loss protocol in healthy individuals have provided evidence that the consolidation of spatial memories, as acquired through navigating and orienteering in spatial surroundings, is negatively affected by total sleep loss. Here, we used both objective and subjective measures to characterize individuals' quality of sleep, and grouped participants into either a poor (insomnia-like) or normal (control) sleep quality group. We asked participants to solve a wayfinding task in a virtual environment, and scored their performance by measuring the time spent to reach a target location and the number of wayfinding errors made while navigating. We found that participants with poor sleep quality were slower and more error-prone than controls in solving the task. These findings provide novel evidence that pre-existing sleep deficiencies in otherwise healthy individuals affects negatively the ability to learn novel routes, and suggest that sleep quality should be accounted for among healthy individuals performing experimental spatial orientation tasks in virtual environments

Does Locoregional Chemotherapy Still Matter in the Treatment of Advanced Pelvic Melanoma?

Pelvic Melanoma relapse occurs in 15% of patients with loco regional metastases, and 25% of cases do not respond to new target-therapy and/or immunotherapy. Melphalan hypoxic pelvic perfusion may, therefore, be an option for these non-responsive patients. Overall median survival time (MST), stratified for variables, including BRAF V600E mutation and eligibility for treatments with new immunotherapy drugs, was retrospectively assessed in 41 patients with pelvic melanoma loco regional metastases. They had received a total of 175 treatments with Melphalan hypoxic perfusion and cytoreductive excision. Among the 41 patients, 22 (53.7%) patients exhibited a wild-type BRAF genotype, 11 of which were not eligible for immunotherapy. The first treatment resulted in a 97.5% response-rate in the full cohort and a 100% response-rate in the 22 wild-type BRAF patients. MST was 18 months in the full sample, 20 months for the 22 wild-type BRAF patients and 21 months for the 11 wild-type BRAF patients not eligible for immunotherapy. Melphalan hypoxic perfusion is a potentially effective treatment for patients with pelvic melanoma loco regional metastases that requires confirmation in a larger multicenter study

Circulating tumour cell gene expression and chemosensitivity analyses: predictive accuracy for response to multidisciplinary treatment of patients with unresectable refractory recurrent rectal cancer or unresectable refractory colorectal cancer liver metastases

Abstract Background Patients with unresectable recurrent rectal cancer (RRC) or colorectal cancer (CRC) with liver metastases, refractory to at least two lines of traditional systemic therapy, may receive third line intraarterial chemotherapy (IC) and targeted therapy (TT) using drugs selected by chemosensitivity and tumor gene expression analyses of liquid biopsy-derived circulating tumor cells (CTCs). Methods In this retrospective study, 36 patients with refractory unresectable RRC or refractory unresectable CRC liver metastases were submitted for IC and TT with agents selected by precision oncotherapy chemosensitivity assays performed on liquid biopsy-derived CTCs, transiently cultured in vitro, and by tumor gene expression in the same CTC population, as a ratio to tumor gene expression in peripheral mononuclear blood cells (PMBCs) from the same individual. The endpoint was to evaluate the predictive accuracy of a specific liquid biopsy precision oncotherapy CTC purification and in vitro culture methodology for a positive RECIST 1.1 response to the therapy selected. Results Our analyses resulted in evaluations of 94.12% (95% CI 0.71–0.99) for sensitivity, 5.26% (95% CI 0.01–0.26) for specificity, a predictive value of 47.06% (95% CI 0.29–0.65) for a positive response, a predictive value of 50% (95% CI 0.01–0.98) for a negative response, with an overall calculated predictive accuracy of 47.22% (95% CI 0.30–0.64). Conclusions This is the first reported estimation of predictive accuracy derived from combining chemosensitivity and tumor gene expression analyses on liquid biopsy-derived CTCs, transiently cultured in vitro which, despite limitations, represents a baseline and benchmark which we envisage will be improve upon by methodological and technological advances and future clinical trials

Cognitive and Emotional Empathy in Individuals with Spinal Cord Injury

Background. Empathy has been conceptualized as comprising a cognitive and an emotional component, the latter being further divided into direct and indirect aspects, which refer, respectively, to the explicit evaluation of the observer’s feelings while attending someone in an emotional situation and to the physiological response of the observer. Empathy has been previously investigated in several neurological disorders. Objective. This study is aimed at investigating empathy in patients with spinal cord injury (SCI). We hypothesize that, due to deafferentation following their injury, SCI patients will display difficulty in the processing of emotional stimuli and blunted empathic responses as compared to healthy controls. Materials and Methods. 20 patients with spinal cord injury (SCI) (12 males and 8 females, mean age=50.9, standard deviation SD=16.1 years; mean education=10.9, SD=4.1 years) were included in the study and compared to 20 matched healthy subjects. Participants were investigated using the State-Trait Anxiety Inventory (Form Y) (STAI-Y), the Beck Depression Scale, and the Toronto Alexithymia Scale. Moreover, participants were further evaluated by means of the Interpersonal Reactivity Index (IRI), which explores both cognitive and emotional aspects of empathy, and through an experimental protocol based on the use of a modified version of the computerized Multifaceted Empathy Test (MET) to evaluate emotional (direct and indirect) empathy and the ability to judge the valence of complex emotional scenes. Results. As compared to healthy controls, SCI patients reported higher scores on the Perspective-Taking subscale of the IRI, while, on the modified MET, they were less accurate in identifying the valence of neutral scenes, notwithstanding their spared direct and indirect emotional empathy ability. Furthermore, we found a significant negative correlation between the time interval since injury and the direct emotional empathy scores on the positive images, as well as a negative correlation with the indirect emotional empathy scores on both positive and neutral images, indicating a blunting of the empathic responses as time elapses. Conclusion. Results suggest that SCI patients, when analyzing the meaning of emotional stimuli, tend to rely on a cognitive empathy strategy rather than on emotion simulation

RFID as a new ICT tool to monitor specimen life cycle and quality control in a biobank

Background: Biospecimen quality is crucial for clinical and translational research and its loss is one of the main obstacles to experimental activities. Beside the quality of samples, preanalytical variations render the results derived from specimens of different biobanks or even within the same biobank incomparable. Specimens collected along the years should be managed with a heterogeneous life cycle. Hence, we propose to collect detailed data concerning the whole life cycle of stored samples employing radio-frequency identification (RFID) technology. Methods: We describe the processing chain of blood biosamples that is operative at the biobank of IRCSS San Raffaele, Rome, Italy (BioBIM). We focus on the problem of tracing the stages following automated preanalytical processing: we collected the time stamps of all events that could affect the biological quality of the specimens by means of RFID tags and readers. Results: We developed a pilot study on a fragment of the life cycle, namely the storage between the end of the preanalytics and the beginning of the analytics, which is usually not traced by automated tools because it typically includes manual handling. By adopting RFID devices we identified the possible critical time delays. At 1, 3 and 6 months RFID-tagged specimens cryopreserved at -80 degrees C were successfully read. Conclusions: We were able to record detailed information about the storage phases and a fully documented specimen life cycle. This will allow us to promote and tune up the best practices in biobanking because i) it will be possible to classify sample features with a sharper resolution, which allows future utilization of stored material; ii) cost-effective policies can be adopted in processing, storing and selecting specimens; iii) after using each aliquot, we can study the life cycle of the specimen with a possible feedback on the procedures

Multidisciplinary Treatment, Including Locoregional Chemotherapy, for Merkel-Polyomavirus-Positive Merkel Cell Carcinomas: Perspectives for Patients Exhibiting Oncogenic Alternative Δ exon 6–7 TrkAIII Splicing of Neurotrophin Receptor Tropomyosin-Related Kinase A

Merkel cell carcinomas (MCCs) are rare, aggressive, cutaneous neuroendocrine tumours, approximately 80% of which are caused by the genomic integration of Merkel cell polyomavirus (MCPyV). MCPyV-positive MCCs carry poor prognosis in approximately 70% of cases, highlighting the need for greater understanding of the oncogenic mechanisms involved in pathogenesis, progression and post-therapeutic relapse, and translation into novel therapeutic strategies. In a previous pilot study, we reported a potential relationship between MCPyV gene expression and oncogenic alternative Δ exon 6–7 TrkAIII splicing in formalin-fixed paraffin-embedded (FFPE) MCC tissues from a 12-patient cohort of >90% MCPyV-positive MCCs, diagnosed at San Salvatore Hospital, L’Aquila, Italy, characterising a new MCC subgroup and unveiling a novel potential MCPyV oncogenic mechanism and therapeutic target. This, however, could not be fully verified due to poor RNA quality and difficulty in protein extraction from FFPE tissues. Here, therefore, we extend our previous observations to confirm the relationship between MCPyV and oncogenic alternative Δ exon 6–7 TrkAIII splicing in fresh, nonfixed, MCPyV-positive MCC metastasis by detecting sequence-verified RT-PCR products, including full-length Δ exon 6–7 TrkAIII, and by Western blot detection of a 100 kDa TrkA protein isoform of identical size to 100 kDa Δ exon 6–7 TrkAIII expressed by stable transfected SH-SY5Y cells. We also report that in three MCC patients submitted for multidisciplinary treatment, including locoregional chemotherapy, MCPyV large T-antigen mRNA expression, Δ exon 6–7 TrkAIII mRNA expression and intracellular indirect immunofluorescence (IF) TrkA and phosphorylation protein isoform(s) immunoreactivity in FFPE tissues were not reduced in postchemotherapeutic-relapsed MCCs compared to pretherapeutic MCCs, extending the possible roles of this novel potential MCPyV oncogenic mechanism from MCC pathogenesis to post-therapeutic relapse and progression. Detection of alternative Δ exon 6–7 TrkAIII splicing in MCC, therefore, not only characterises a new MCPyV-positive MCC subgroup and unveils a novel potential MCPyV oncogenic mechanism but also identifies patients who may benefit from inhibitors of MCPyV T-antigen and/or TrkAIII expression or clinically approved Trk kinase inhibitors such as larotrectinib or entrectinib, which are known to inhibit activated TrkA oncogenes and to elicit durable responses in TrkA-fusion oncogene-driven cancers, supporting the call for a large-scale multicentre clinical study

The Alternative TrkAIII Splice Variant, a Targetable Oncogenic Participant in Human Cutaneous Malignant Melanoma

Post-therapeutic relapse, poor survival rates and increasing incidence justify the search for novel therapeutic targets and strategies in cutaneous malignant melanoma (CMM). Within this context, a potential oncogenic role for TrkA in CMM is suggested by reports of NTRK1 amplification, enhanced TrkA expression and intracellular TrkA activation associated with poor prognosis. TrkA, however, exhibits tumour-suppressing properties in melanoma cell lines and has recently been reported not to be associated with CMM progression. To better understand these contradictions, we present the first analysis of potential oncogenic alternative TrkA mRNA splicing, associated with TrkA immunoreactivity, in CMMs, and compare the behaviour of fully spliced TrkA and the alternative TrkAIII splice variant in BRAF(V600E)-mutated A375 melanoma cells. Alternative TrkA splicing in CMMs was associated with unfolded protein response (UPR) activation. Of the several alternative TrkA mRNA splice variants detected, TrkAIII was the only variant with an open reading frame and, therefore, oncogenic potential. TrkAIII expression was more frequent in metastatic CMMs, predominated over fully spliced TrkA mRNA expression in ≈50% and was invariably linked to intracellular phosphorylated TrkA immunoreactivity. Phosphorylated TrkA species resembling TrkAIII were also detected in metastatic CMM extracts. In A375 cells, reductive stress induced UPR activation and promoted TrkAIII expression and, in transient transfectants, promoted TrkAIII and Akt phosphorylation, enhancing resistance to reductive stress-induced death, which was prevented by lestaurtinib and entrectinib. In contrast, fully spliced TrkA was dysfunctional in A375 cells. The data identify fully spliced TrkA dysfunction as a novel mechanism for reducing melanoma suppression, support a causal relationship between reductive stress, UPR activation, alternative TrkAIII splicing and TrkAIII activation and characterise a targetable oncogenic pro-survival role for TrkAIII in CMM

Cognitive Effects of Repeated Acute Exposure to Very High Altitude Among Altitude-Experienced Workers at 5050 m

Background: We investigated altitude effects on different cognitive domains among perennial shift-workers at the Atacama Large Millimeter/submillimeter Array Observatory (5050 m), Chile. Materials and Methods: Twenty healthy male workers were recruited and assigned to either a moderate-altitude first (MAF group, Test 1: 2900 m and Test 2: 5050 m) or to a high-altitude first (HAF group, Test 1: 5050 m and Test 2: 2900 m). Test 1 was conducted at the beginning and Test 2 at the end of the shift-work week. Processing speed (RTI, reaction time), attention (AST, attention-switching task, and RVP, rapid visual processing), and executive function (OTS, One Touch Stockings of Cambridge) were assessed. Results: Of the three cognitive domains assessed, only processing speed showed altitude-at-test group interaction (RTI median five choice reaction time: F1, 17 = 6.980,  = 0.291, p = 0.017). With acclimatization, there was a decrease in AST reaction latency mean (t17 = −2.155, dz = 1.086, p = 0.046), an increase in RVP accuracy (t17 = 2.733, dz = 1.398, p = 0.014), and a decrease in OTS mean latency first choice (t17 = −2.375, dz = 1.211, p = 0.03). Decreased variability in cognitive function was observed in AST reaction latency standard deviation (t17 = −2.524, dz = 1.282, p = 0.022) and in RVP response latency standard deviation (t17 = −2.35, dz = 1.177, p = 0.03) with acclimatization. At 5050 m of elevation, SpO2 was positively correlated with executive function in the MAF group (OTS problems solved on first choice: r(5) = 0.839, p = 0.018) and negatively correlated with executive function latency standard deviations in the HAF group (OTS latency to first choice standard deviation: r(10) = −0.618, p = 0.032). Conclusions: Our findings highlight the importance of acclimatization and improvement of blood oxygen level, even among high altitude-experienced workers, to optimize performance of cognitively demanding work and reduce high altitude-associated health risks