Unveiling nuclear chromatin distribution using IsoConcentraChromJ: A flourescence imaging plugin for IsoRegional and IsoVolumetric based ratios analysis.

Chromatin exhibits non-random distribution within the nucleus being arranged into discrete domains that are spatially organized throughout the nuclear space. Both the spatial distribution and structural rearrangement of chromatin domains in the nucleus depend on epigenetic modifications of DNA and/or histones and structural elements such as the nuclear envelope. These components collectively contribute to the organization and rearrangement of chromatin domains, thereby influencing genome architecture and functional regulation. This study develops an innovative, user-friendly, ImageJ-based plugin, called IsoConcentraChromJ, aimed quantitatively delineating the spatial distribution of chromatin regions in concentric patterns. The IsoConcentraChromJ can be applied to quantitative chromatin analysis in both two- and three-dimensional spaces. After DNA and histone staining with fluorescent probes, high-resolution images of nuclei have been obtained using advanced fluorescence microscopy approaches, including confocal and stimulated emission depletion (STED) microscopy. IsoConcentraChromJ workflow comprises the following sequential steps: nucleus segmentation, thresholding, masking, normalization, and trisection with specified ratios for either 2D or 3D acquisitions. The effectiveness of the IsoConcentraChromJ has been validated and demonstrated using experimental datasets consisting in nuclei images of pre-adipocytes and mature adipocytes, encompassing both 2D and 3D imaging. The outcomes allow to characterize the nuclear architecture by calculating the ratios between specific concentric nuclear areas/volumes of acetylated chromatin with respect to total acetylated chromatin and/or total DNA. The novel IsoConcentrapChromJ plugin could represent a valuable resource for researchers investigating the rearrangement of chromatin architecture driven by epigenetic mechanisms using nuclear images obtained by different fluorescence microscopy methods.

The Anti-Inflammatory Potential of an Ethanolic Extract from Sarcopoterium spinosum Fruits for Protection and/or Counteraction against Oxidative Stress in Dysfunctional Endothelial Cells.

Plants and plant extracts are a relevant source of bioactive compounds widely employed as functional foods. In the Mediterranean area, the shrub Sarcopoterium spinosum is traditionally used as an herbal medicine for weight loss and a diabetes treatment. Inflammation is a protective mechanism involved in the development of many pathological conditions, including cardiovascular diseases. The present study aimed to investigate in vitro the antioxidant and cytoprotective properties of an ethanolic extract from S. spinosum fruits (SEE) in a cellular model of endothelium dysfunction. Corilagin and quercetin are two polyphenols abundant in SEE and were tested for comparison. The exposure of HECV cells for 24 h to 30 µM hydrogen peroxide (H2O2) lead to an oxidative stress condition. When HECV cells were treated with 10 µg/mL of SEE or single compounds after or before the oxidative insult, the results showed their ability to (i) decrease the reactive oxygen species (ROS) production quantified using fluorometric analysis and the lipid peroxidation measured with a spectrophotometric assay; (ii) rescue both the glutathione reduced to oxidized (GSH/GSSG) ratio and nitric oxide impair and the protein denaturation; and (iii) accelerate the wound repair measured using a T-scratch assay. Taken together, our findings indicate that the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical application.

The Sound of Grief: A Critical Discussion on the Experience of Creating and Listening to the Digitally Reproduced Voice of the Deceived.

Technological tools allow for the reproduction and control of peculiar stimuli, such as the possibility of producing audio clips with the voices of deceased people. Artificial intelligence allows to create at-home vocal messages from an audioclip. Recently, some videos and documentaries depicting people interacting with artificial intelligence content related to the deceased have been released to the general public. However, the possibility of interacting with realistic stimuli related to deceased loved ones can create peculiar and delicate experiences and should gain the attention of the scientific community and mental health professionals. Listening and searching for experiences related to the deceived ones might indicate a natural way to elaborate and live the experience of grieving or the presence of symptoms related to more severe conditions. Moreover, such powerful stimuli might be potentially harmful to users, if not appropriately used. To the best of our knowledge, no scientific literature exists on the topic of listening to audio clips with the voice of the deceased yet, although various people shared thoughts and feelings about these habits on social networks and forums. Given the relevant psychological impact that grief can have on a person, an open discussion on the possibility and risks of the availability of digital stimuli related to grief should be taken into account by the scientific community.

Polyphenol-enriched extracts of Sarcopoterium spinosum fruits for counteracting lipid accumulation and oxidative stress in an in vitro model of hepatic steatosis.

Sarcopoterium spinosum (L.) Spach is a Rosaceae shrub employed in the folk medicine in the Eastern Mediterranean basin. The previous few studies have focused on the S. spinosum roots, while the fruits have been poorly investigated. The present study aims to assess the biological properties of S. spinosum fruits collected in Lebanon and subjected to ethanolic, water or boiling water extraction. The extracts were compared for the phenol and flavonoid contents, and for the in vitro radical scavenging ability. The ethanolic extract (SEE) was selected and characterized by high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS) showing a phenolome rich in tannins (ellagitannins), flavonoids (quercetin derivatives), and triterpenes. The biological activity of SEE was tested on a cellular model of moderate steatosis consisting of lipid-loaded hepatic cells treated with increasing concentrations of SEE (1-25 µg/mL), or with corilagin or quercetin as comparison. In steatotic hepatocytes the SEE was able (i) to ameliorate the hepatosteatosis; (ii) to counteract the excess ROS and lipid peroxidation; (iii) to restore the impaired catalase activity. The results indicate that the ethanolic extract from S. spinosum fruits is endowed with relevant antisteatotic and antioxidant activities and might find application as nutraceutical product.

Towards an Informed CNN for Bone SR-microCT Image Classification with an Unsupervised Patched-based Image Clustering.

Bone microscale differences cannot be readily recognizable to humans from Synchrotron Radiation micro-Computed Tomography (SR-microCT) images. Premises are possible with Deep Learning (DL) imaging analysis. Despite this, more attention to high-level features leads models to require help identifying relevant details to support a decision. Within this context, we propose a method for classifying healthy, osteoporotic, and COVID-19 femoral heads SR-microCT images informing a vgg16 about the most subtle microscale differences using unsupervised patched-based clustering. Our strategy allows achieving up to 9.8% accuracy improvement in classifying healthy from osteoporotic images over uninformed methods, while 59.1% of accuracy between osteoporosis and COVID-19.Clinical relevance-We established a starting point for classifying healthy, osteoporotic, and COVID-19 femoral heads from SR-microCTs with human non-discriminative features, with 60.91% accuracy in healthy-osteporotic image classification.

Nuclear and chromatin rearrangement associate to epigenome and gene expression changes in a model of in vitro adipogenesis and hypertrophy.

Hypertrophy of adipocytes represents the main cause of obesity. We investigated in vitro the changes associated with adipocyte differentiation and hypertrophy focusing on the nuclear morphometry and chromatin epigenetic remodelling. The 3 T3-L1 pre-adipocytes were firstly differentiated into mature adipocytes, then cultured with long-chain fatty acids to induce hypertrophy. Confocal and super-resolution stimulation emission depletion (STED) microscopy combined with ELISA assays allowed us to explore nuclear architecture, chromatin distribution and epigenetic modifications. In each condition, we quantified the triglyceride accumulation, the mRNA expression of adipogenesis and dysfunction markers, the release of five pro-inflammatory cytokines. Confocal microscopy revealed larger volume and less elongated shape of the nuclei in both mature and hypertrophic cells respect to pre-adipocytes, and a trend toward reduced chromatin compaction. Compared to mature adipocytes, the hypertrophic phenotype showed larger triglyceride content, increased PPARγ expression reduced IL-1a release, and up-regulation of a pool of genes markers for adipose tissue dysfunction. Moreover, a remodelling of both epigenome and chromatin organization was observed in hypertrophic adipocytes, with an increase in the average fluorescence of H3K9 acetylated domains in parallel with the increase in KAT2A expression, and a global hypomethylation of DNA. These findings making light on the nuclear changes during adipocyte differentiation and hypertrophy might help the strategies for treating obesity and metabolic complications.

Osteoporosis and Covid-19: Detected similarities in bone lacunar-level alterations via combined AI and advanced synchrotron testing.

While advanced imaging strategies have improved the diagnosis of bone-related pathologies, early signs of bone alterations remain difficult to detect. The Covid-19 pandemic has brought attention to the need for a better understanding of bone micro-scale toughening and weakening phenomena. This study used an artificial intelligence-based tool to automatically investigate and validate four clinical hypotheses by examining osteocyte lacunae on a large scale with synchrotron image-guided failure assessment. The findings indicate that trabecular bone features exhibit intrinsic variability related to external loading, micro-scale bone characteristics affect fracture initiation and propagation, osteoporosis signs can be detected at the micro-scale through changes in osteocyte lacunar features, and Covid-19 worsens micro-scale porosities in a statistically significant manner similar to the osteoporotic condition. Incorporating these findings with existing clinical and diagnostic tools could prevent micro-scale damages from progressing into critical fractures.

Isolating the Role of Bone Lacunar Morphology on Static and Fatigue Fracture Progression through Numerical Simulations.

Currently, the onset of bone damage and the interaction of cracks with the surrounding micro-architecture are still black boxes. With the motivation to address this issue, our research targets isolating lacunar morphological and densitometric effects on crack advancement under both static and cyclic loading conditions by implementing static extended finite element models (XFEM) and fatigue analyses. The effect of lacunar pathological alterations on damage initiation and progression is evaluated; the results indicate that high lacunar density considerably reduces the mechanical strength of the specimens, resulting as the most influencing parameter among the studied ones. Lacunar size has a lower effect on mechanical strength, reducing it by 2%. Additionally, specific lacunar alignments play a key role in deviating the crack path, eventually slowing its progression. This could shed some light on evaluating the effects of lacunar alterations on fracture evolution in the presence of pathologies.

Adipocyte-hepatocyte crosstalk in cellular models of obesity: Role of soluble factors.

Hepatic steatosis is often a consequence of obesity. Adipose tissue is an important endocrine regulator of metabolic homeostasis in the body. In obesity, adipocytes become hypertrophic and develop an inflammatory phenotype, altering the panel of secreted adipokines. Moreover, excess fatty acids are, in part, released by adipocytes and delivered to the liver. These multiple pathways of adipose-liver crosstalk contribute to the development and progression of liver disease: TNFα induces hepatocyte dysfunction, excess of circulating fatty acids promotes hepatic steatosis and inflammation, whilst adipokines mediate and exacerbate liver injury. In this study, we investigated in vitro the effects and mechanisms of the crosstalk between adipocytes and hepatocytes, as a function of the different adipocyte status (mature vs hypertrophic) being mediated by soluble factors. We employed the conditioned medium method to test how mature and hypertrophic adipocytes distinctively affect the liver, leading to metabolic dysfunction. The media collected from adipocytes were characterized by high triglyceride content and led to lipid accumulation and fat-dependent dysfunction in hepatocytes. The present findings seem to suggest that, in addition to triglycerides, other soluble mediators, cytokines, are released by mature and hypertrophic adipocytes and influence the metabolic status of liver cells. Understanding the precise factors involved in the pathogenesis and pathophysiology of NAFLD in obesity will provide important insights into the mechanisms responsible for the metabolic complications of obesity, paving the way for new possible approaches.

How to Measure Propensity to Take Risks in the Italian Context: The Italian Validation of the Risk Propensity Scale.

Risk propensity is a multifaced construct that influences many aspects of life, such as decision making. In the present study, the psychometric characteristics of the Risk Propensity Scale (RPS) have been explored for the first time in an Italian sample. The RPS is a 7 item self-report questionnaire measuring people's tendency to take risks. The English RPS has been translated following the forward-backwards translation method, and it was filled out by 199 participants. Since its dimensionality has never been explored before, its factor structure has been analysed with exploratory factor analysis that confirmed the one-factor structure of the questionnaire and the retention of all the items. The Italian version of the RPS has high internal consistency (Cronbach alphas .78), and almost all the items were positively and significantly correlated. The convergent and discriminant validity, analysed by considering the associations with decision-making styles and an implicit measure of risk propensity, were satisfactory. Overall, the Italian version of the RPS is a valid and quick questionnaire useful to measure propensity to take risks in the Italian context.

From virtual to real healing: a critical overview of the therapeutic use of virtual reality to cope with mourning.

In recent years, virtual reality (VR) has been effectively employed in several settings, ranging from health care needs to leisure and gaming activities. A new application of virtual stimuli appeared in social media: in the documentary 'I met you' from the South-Korean Munhwa Broadcasting, a mother made the experience of interacting with the avatar of the seven-year-old daughter, who died four years before. We think that this new application of virtual stimuli should open a debate on its possible implications: it represents contents related to grief, a dramatic and yet natural experience, that can have deep psychological impacts on fragile subjects put in virtual environments. In the present work, possible side-effects, as well as hypothetical therapeutical application of VR for the treatment of mourning, are discussed.

The synergy of synchrotron imaging and convolutional neural networks towards the detection of human micro-scale bone architecture and damage.

The growing health and economic burden of bone fractures, their intricate multiscale features and the existing knowledge gaps in the comprehension of micro-scale bone damage occurrence make fracture diagnosis a challenging issue. In this scenario, deep-learning and artificial intelligence embody the new frontier of healthcare system, by overcoming the subjectivity of clinicians in the analysis of medical images. However, the preliminary attempts in exploiting the power of machine learning algorithms such as neural networks are still limited to bone macro-scale, while there is an evident lack in their application to smaller scales, where damage starts nucleating. Currently, speculations at the micro-scale are only feasible with the aid of high-resolution imaging techniques, that are particularly time consuming in terms of output images analysis. In this context, this works aims at combining the visualization of the micro-crack propagation mechanism with the promising application of convolutional neural networks. The implemented artificial intelligence tool is based for the first time on a large number of human synchrotron images coming from healthy and osteoporotic femoral heads tested under micro-compression. The designed convolutional neural networks are able to automatically detect lacunae and micro-cracks at different compression levels with high accuracy levels; indeed, with the baseline setup, networks achieve more than 0.99 level of accuracy for both cracks and lacunae, and more than 0.87 of the meanIoU adopted as validation metric. This approach is particularly encouraging for the development of powerful recognition system to comprehend bone micro-damage initiation and propagation, paving the way to the application of machine learning studies to bone micromechanics. This could be additionally crucial for future patient specific micro-scale observations to be related to the clinical practice.

Influence of Simulated In Vitro Gastrointestinal Digestion on the Phenolic Profile, Antioxidant, and Biological Activity of Thymbra spicata L. Extracts.

Plants or plant extracts are widely investigated for preventing/counteracting several chronic disorders. The oral route is the most common route for nutraceutical and drug administration. Currently, it is still unclear as to whether and how the pattern of phenolic compounds (PCs) found in the plants as well as their bioactivity could be modified during the gastrointestinal transit. Recent studies have revealed antioxidant and anti-steatotic properties of Thymbra spicata. Here, we investigated the possible loss of phytochemicals that occurs throughout the sequential steps of a simulated in vitro gastrointestinal (GI) digestion of aqueous and ethanolic extracts of aerial parts of T. spicata. Crude, digested, and dialyzed extracts were characterized in terms of their phenolic profile and biological activities. Total contents of carbohydrates, proteins, PCs, flavonoids, and hydroxycinnamic acids were quantified. The changes in the PC profile and in bioactive compounds upon the simulated GI digestion were monitored by HPLC-MS/MS analysis. The antioxidant activity was measured by different spectrophotometric assays, and the antiproliferative potential was assessed by using three representative human cancer cell lines. We observed that the simulated GI digestion reduced the phytochemical contents in both aqueous and ethanolic T. spicata extracts and modified the PC profile. However, T. spicata extracts improved their antioxidant potential after digestion, while a partial reduction in the antiproliferative activity was observed for the ethanolic extract. Therefore, our results could provide a scientific basis for the employment of T. spicata extract as valuable nutraceutical.

The Potential of Lamiaceae Herbs for Mitigation of Overweight, Obesity, and Fatty Liver: Studies and Perspectives.

Numerous plants, plant extracts, and plant-derived compounds are being explored for their beneficial effects against overweight and liver diseases. Obesity is associated with the increased prevalence of non-alcoholic fatty liver disease (NAFLD), becoming the most common liver disease in Western countries. Obesity and NAFLD are closely associated with many other metabolic alternations such as insulin resistance, diabetes mellitus, and cardiovascular diseases. Many herbs of the Lamiaceae family are widely employed as food and spices in the Mediterranean area, but also in folk medicine, and their use for the management of metabolic disorders is well documented. Hereby, we summarized the scientific results of the medicinal and nutraceutical potential of plants from the Lamiaceae family for prevention and mitigation of overweight and fatty liver. The evidence indicates that Lamiaceae plants may be a cost-effective source of nutraceuticals and/or phytochemicals to be used in the management of metabolic-related conditions such as obesity and NAFLD. PubMed, Google Scholar, Scopus, and SciFinder were accessed to collect data on traditional medicinal plants, compounds derived from plants, their reported anti-obesity mechanisms, and therapeutic targets.

The Involvement of Polyamines Catabolism in the Crosstalk between Neurons and Astrocytes in Neurodegeneration.

In mammalian cells, the content of polyamines is tightly regulated. Polyamines, including spermine, spermidine and putrescine, are involved in many cellular processes. Spermine oxidase specifically oxidizes spermine, and its deregulated activity has been reported to be linked to brain pathologies involving neuron damage. Spermine is a neuromodulator of a number of ionotropic glutamate receptors and types of ion channels. In this respect, the Dach-SMOX mouse model overexpressing spermine oxidase in the neocortex neurons was revealed to be a model of chronic oxidative stress, excitotoxicity and neuronal damage. Reactive astrocytosis, chronic oxidative and excitotoxic stress, neuron loss and the susceptibility to seizure in the Dach-SMOX are discussed here. This genetic model would help researchers understand the linkage between polyamine dysregulation and neurodegeneration and unveil the roles of polyamines in the crosstalk between astrocytes and neurons in neuroprotection or neurodegeneration.

Medical Students and Professionals Facing the COVID-19 Pandemic: A Cross-Sectional Survey Study about Similarities and Differences.

This study aimed at exploring the emotional reaction that medical students (MS) and professionals have faced during the COVID-19 pandemic and investigating the differences between the two groups. A total of 362 MS and 330 professionals filled in an online survey during the second outbreak of the COVID-19 pandemic in Italy. The outcome measures were psychological distress with the General Health Questionnaire, stress, fear for themselves, fear for family members and cohabitants, perceived control, anger, loneliness, and feeling abandoned by institutions with Visual Analog Scales (VAS) and two open-ended questions about their perceived difficulties and the perceived consequences of the pandemic. The results showed that the level of distress among the two groups was above the threshold (MS mean: 21.85; professionals mean: 21.25). The means of the VAS of MS and professionals showed different results for the two groups, and we analyzed them with independent samples t-tests and linear regressions. MS reported higher levels of perceived loneliness (t(673.177) = −1.970, p < 0.05), while physicians reported more fear for themselves (t(655.183) = 4.799, p < 0.001), anger (t(690) = 2.096, p < 0.05), and feeling abandoned by the institutions (t(690) = 7.296, p < 0.001). The open questions were analyzed considering the most frequent words used to describe their perceived difficulties and consequences; the specificity analysis emphasizes the differences in the words used by the two groups. In conclusion, MS and physicians reported similar levels of psychological symptoms. Physicians were mostly worried about themselves, they were afraid of getting infected, while MS suffered from loneliness and the missed possibilities concerning their education.

Relationship between Liver Stiffness and Steatosis in Obesity Conditions: In Vivo and In Vitro Studies.

Obesity is a major risk factor for metabolic dysfunction such as non-alcoholic fatty liver disease (NAFLD). The NAFLD spectrum ranges from simple steatosis, to steatohepatitis, fibrosis, and cirrhosis. The aim of this study is to characterize the grade of steatosis being associated with overnutrition and obesity, both at the level of single hepatocyte and whole liver, and to correlate it with the hepatocyte/liver stiffness and dysfunction. For the in vivo study, 60 subjects were enrolled and grouped based on the stage of liver steatosis/fibrosis according to biochemical analyses, liver ultrasonography (USG) and acoustic radiation force impulse shear wave elastography (ARFI-SWE). For single hepatocyte analyses we employed in vitro models of moderate and severe steatosis on which to assess the single cell biomechanics by Single Cell Force Spectroscopy (SCFS) and Quantitative Phase Microscopy (QPM). Results show that in vivo liver stiffness depends mainly on the extent of fat accumulation and not on fibrosis. These results parallel the in vitro observations showing that hepatocyte stiffness and dysfunction increase with increasing fat accumulation and lipid droplet enlargement. Our findings indicate that the extent of steatosis markedly affects the biomechanical properties of both liver and single hepatocytes thus proving insights about the role of modulation of liver/hepatocyte elasticity as a physical mechanism transducing the obesity-dependent excess of plasmatic lipids towards liver steatosis and dysfunction.

Mechanical Design Optimization of Prosthetic Hand's Fingers: Novel Solutions towards Weight Reduction.

From the mechanical function of grabbing objects to the emotional aspect of gesturing, the functionality of human hands is fundamental for both physical and social survival. Therefore, the loss of one or both hands represents a devastating issue, exacerbated by long rehabilitation times and psychological treatments. Prosthetic arms represent an effective solution to provide concrete functional and esthetical support. However, commercial hand prostheses still lack an optimal combination of light weight, durability, adequate cosmetic appearance, and affordability. Among these aspects, the priority for upper-limb prosthesis users is weight, a key parameter that influences both the portability and the functionality of the system. The purpose of this work is to optimize the design of the MyHand prosthesis, by redesigning both the proximal and distal finger and thumb in light of finding an optimal balance between weight reduction and adequate stiffness. Starting from elastic-plastic numerical models and experimental tests on obsolete components, analyzed under the worst loading condition, five different design solutions are suggested. An iterative topology optimization process locates the regions where material removal is permitted. From these results, 2 mm geometrical patterns on the top surface of the hand prosthesis appear as the most prominent, preventing object intrusion.

Beneficial Effects of Carvacrol on In Vitro Models of Metabolically-Associated Liver Steatosis and Endothelial Dysfunction: A Role for Fatty Acids in Interfering with Carvacrol Binding to Serum Albumin.

BACKGROUND: Carvacrol, a plant phenolic monoterpene, is largely employed as food additive and phytochemical. OBJECTIVE: We aimed to assess the lipid lowering and protective effects of carvacrol in vitro using cellular models of hepatic steatosis and endothelial dysfunction. We also investigated if and how the binding of carvacrol to albumin, the physiological transporter for small compounds in the blood, might be altered by the presence of high levels of fatty acids (FAs). METHODS: Hepatic FaO cells treated with exogenous FAs mimic hepatosteatosis; endothelial HECV cells exposed to hydrogen peroxide are a model of endothelial dysfunction. In these models, we measured spectrophotometrically lipid accumulation and release, lipoperoxidation, free radical production, and nitric oxide release before and after treatment with carvacrol. The carvacrol binding to albumin in the presence or absence of high levels of FAs was assessed by absorption and emission spectroscopies. RESULTS: Carvacrol counteracted lipid accumulation and oxidative stress in hepatocytes and protected endothelial cells from oxidative stress and dysfunction. Moreover, high levels of FAs reduced the binding of carvacrol to albumin. CONCLUSION: The results suggest the good potential of carvacrol in ameliorating dysfunction of hepatic and endothelial cells in vitro. High levels of circulating FAs might compete with carvacrol for binding to albumin thus influencing its transport and bio-distribution.

After the Pandemic: The Future of Italian Medicine. The Psychological Impact of COVID-19 on Medical and Other Healthcare-Related Degrees Students.

Objective: The study aimed to explore the psychological symptoms and the readiness to fight the pandemic of the new generation of healthcare professionals: medical and other healthcare degree students. Methods: We enrolled 509 medical and healthcare-related degree students during the second outbreak of COVID-19 in Italy. We have examined their psychological symptoms using the 12-item General Health Questionnaire (GHQ-12) and their readiness to fight the pandemic together with their academic career status, their relationship with the university, and their emotional reactions to the pandemic with Visual Analog Scales. Results: We retrieved a GHQ mean of 21.65 (SD = 40.63) and readiness to fight the pandemic mean of 53.58 (SD = 31.49). Perceived control affects variables: a negative effect on psychological symptoms and a positive effect on the willingness to fight the pandemic. The other variables with an impact were stress, loneliness, and anger that had a significant and positive impact on psychological symptoms. Age and concern for patients had a significant positive impact on readiness to fight for the pandemic, while years of attendance had a significant but negative impact. Conclusion: Universities and Institutions should consider the impact of the pandemic on students, in particular, for its effect on their mental health.