Cytotoxicity-related Gene Expression and Chromatin Accessibility Define a Subset of CD4+ T Cells that Mark Progression to Type 1 Diabetes

Type 1 diabetes in children is heralded by a preclinical phase defined by circulating autoantibodies to pancreatic islet antigens. How islet autoimmunity is initiated and then progresses to clinical diabetes remains poorly understood. Only one study has reported gene expression in specific immune cells of at-risk children, associated with progression to islet autoimmunity. We analysed gene expression by RNAseq in CD4⁺ and CD8⁺ T cells, NK cells and B cells, and chromatin accessibility by ATACseq in CD4⁺ T cells, in five genetically at-risk children with islet autoantibodies who progressed to diabetes over a median of 3 years (‘Progressors’) compared to five children matched for sex, age and HLA-DR who had not progressed (‘Non-progressors). In Progressors, differentially expressed genes (DEGs) were largely confined to CD4⁺ T cells and enriched for cytotoxicity-related genes/pathways. Several top-ranked DEGs were validated in a semi-independent cohort of 13 Progressors and 11 Non-progressors. Flow cytometry confirmed progression was associated with expansion of CD4⁺cells with a cytotoxic phenotype. By ATAC-seq, progression was associated with reconfiguration of regulatory chromatin regions in CD4⁺cells, some linked to differentially expressed cytotoxicity-related genes. Our findings suggest that cytotoxic CD4⁺T cells play a role in promoting progression to type 1 diabetes.

Cytotoxicity-related Gene Expression and Chromatin Accessibility Define a Subset of CD4+ T Cells that Mark Progression to Type 1 Diabetes

Type 1 diabetes in children is heralded by a preclinical phase defined by circulating autoantibodies to pancreatic islet antigens. How islet autoimmunity is initiated and then progresses to clinical diabetes remains poorly understood. Only one study has reported gene expression in specific immune cells of at-risk children, associated with progression to islet autoimmunity. We analysed gene expression by RNAseq in CD4⁺ and CD8⁺ T cells, NK cells and B cells, and chromatin accessibility by ATACseq in CD4⁺ T cells, in five genetically at-risk children with islet autoantibodies who progressed to diabetes over a median of 3 years (‘Progressors’) compared to five children matched for sex, age and HLA-DR who had not progressed (‘Non-progressors). In Progressors, differentially expressed genes (DEGs) were largely confined to CD4⁺ T cells and enriched for cytotoxicity-related genes/pathways. Several top-ranked DEGs were validated in a semi-independent cohort of 13 Progressors and 11 Non-progressors. Flow cytometry confirmed progression was associated with expansion of CD4⁺cells with a cytotoxic phenotype. By ATAC-seq, progression was associated with reconfiguration of regulatory chromatin regions in CD4⁺cells, some linked to differentially expressed cytotoxicity-related genes. Our findings suggest that cytotoxic CD4⁺T cells play a role in promoting progression to type 1 diabetes.

Does an Endometrial Cancer Diagnosis among Asymptomatic Patients Improve Prognosis?

Background: Endometrial cancer is the most common gynecological malignancy in developed countries with no screening available. There is still a tendency to provide invasive bioptic verification in asymptomatic women with abnormal ultrasound findings to diagnose carcinoma in a preclinical phase; even though, it is not supported by European guidelines. Our goal was to determine DFS (disease-free survival), OS (overall survival), and DSS (disease-specific survival) differences between symptom-free and symptomatic (bleeding, or spotting) endometrial cancer patients with similar stage and tumor/clinical characteristics. Methods: All of our patients with endometrial cancer following surgical treatment between 2006 and 2019 were assessed, evaluating risk factors for recurrence and death while focusing on bleeding using univariable and multivariable analysis. Results: 625 patients meeting the inclusion criteria were divided into asymptomatic (n = 144, 23%) and symptomatic (n = 481, 77%) groups. The median follow-up was 3.6 years. Using univariable analysis, symptomatic patients had a three times higher risk of recurrence (HR 3.1 (95% Cl 1.24–7.77), p = 0.016). OS (HR 1.35 (0.84–2.19), p = 0.219) and DSS (HR 1.66 (0.64–4.28), p = 0.3) were slightly worse without reaching statistical significance. In our multivariable analysis, symptomatology was deemed completely insignificant in all monitored parameters (DFS: HR 2.03 (0.79–5.24), p = 0.144; OS: HR 0.72 (0.43–1.21), p = 0.216). Conclusions: The symptomatic endometrial cancer patients risk factor of earlier recurrence and death is insignificantly higher when compared with the asymptomatic cohort. However, multivariable analysis verifies that prognosis worsens with other clinically relevant parameters, not by symptomatology itself. In terms of survival outcome in EC patients, we recognized symptomatology as a non-significant marker for the patient’s prognosis.

Medical students positions regarding resource allocation in times of crisis

Objective To compare the perspectives of medical students in the preclinical and clinical phases of medical training on the issue of rationing scarce medical resources in times of crisis. Methods Questionnaire-based cross-sectional study. Results A total of 201 participants took part in the study, with 100 participants in the preclinical phase group, and 101 in the clinical phase group. A multivariable analysis found that just 14.9% (n = 34) of the clinical phase students were willing to give a short-supplied blood unit to the first-arrived patient to the emergency department when more patients are expected compared to 63.9% in the preclinical group (n = 62) ( p < 0.001, OR = 0.75 95% CI: 0.029−0.192). Seventy-four percent (n = 74) of the clinical phase students were found to be willing to remove a patient from a respirator to allocate it to an ill child compared to 35.7% (n = 35) in the preclinical phase group ( p < 0.001, OR = 4.168 95% CI: 1.931−8.998). Of the clinical phase group, 46.6% (n = 41) were willing to allocate a short supplied flu medicine to a patient with poor prognosis compared to 57.7% (n = 56) in the preclinical phase group ( p = 0.04, OR = 0.457 95% CI: 0.216−0.966). Conclusion Clinical exposure during training may affect the way medical students make ethical decisions, independent of age, sex, as well as marital and parental status.

Decreased Heart Rate Variability in COVID-19

On March 12, 2020, the World Health Organization (WHO) announced that the coronavirus disease 2019 (COVID-19) outbreak had become a pandemic. COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which primarily infects the lower airways and binds to Angiotensin-Converting Enzyme 2 (ACE2) on alveolar epithelial cells. ACE2 is widely expressed, not only in the lungs but also in the cardiovascular system. Therefore, SARS-CoV-2 can also damage the myocardium. We analysed three COVID-19 cases that resulted in death and found that either COVID-19 or antiviral drugs could affect the coupling between the autonomic nervous system and the sinus node, thus affecting heart rate variability and preventing the heart rate from rising in response to the increase in body temperature. Early detection of the preclinical phase of cardiac autonomic dysfunction may help determine patients in need of aggressive treatment and control of cardiovascular risk factors. Antiviral drugs should be used with caution in patients with heart injury.

Imaging of the nigrostriatal system for evaluating the preclinical phase of Parkinson’s disease development: the utility of neuromelanin, diffusion MRI, and DAT-SPECT

Objectives: To assess the utility of examining the nigrostriatal system with magnetic resonance imaging (MRI) and dopamine transporter (DAT) imaging for evaluating the preclinical phase of Parkinson’s disease (PD). Methods: The subjects were 32 patients with early PD and a history of probable rapid eye movement sleep behavior disorder (RBD; PD group), 15 patients with idiopathic RBD (RBD group), and 24 age-matched healthy controls (HC group) who underwent neuromelanin and diffusion tensor MRI for analysis of the substantia nigra pars compacta (SNpc). The RBD and PD groups underwent DAT imaging. In the RBD group, totals of 39 MRI and 27 DAT imaging examinations were obtained longitudinally. For each value, intergroup differences and receiver-operating characteristic (ROC) analysis for diagnostic performance were examined statistically. Results: The neuromelanin value was significantly lower and the diffusion tensor values except fractional anisotropy were significantly higher in the RBD and PD groups than in the HC group. The DAT specific binding ratio (SBR) was significantly lower in the PD group than in the RBD group. The areas under the ROC curves (AUCs) for neuromelanin/mean diffusivity value in the SNpc were 0.76/0.82 for diagnosing RBD and 0.83/0.80 for diagnosing PD. The AUC for the SBR for discriminating PD from RBD was 0.87. Conclusions: MRI and DAT imaging may be useful for evaluating sequential nigrostriatal changes during the preclinical phase of PD. Advances in knowledge: MRI detects nigrostriatal changes in both RBD and early PD, and DAT imaging detects nigrostriatal changes during the transition to PD in RBD.

Deep Learning Approaches to Colorectal Cancer Diagnosis: A Review

Unprecedented breakthroughs in the development of graphical processing systems have led to great potential for deep learning (DL) algorithms in analyzing visual anatomy from high-resolution medical images. Recently, in digital pathology, the use of DL technologies has drawn a substantial amount of attention for use in the effective diagnosis of various cancer types, especially colorectal cancer (CRC), which is regarded as one of the dominant causes of cancer-related deaths worldwide. This review provides an in-depth perspective on recently published research articles on DL-based CRC diagnosis and prognosis. Overall, we provide a retrospective synopsis of simple image-processing-based and machine learning (ML)-based computer-aided diagnosis (CAD) systems, followed by a comprehensive appraisal of use cases with different types of state-of-the-art DL algorithms for detecting malignancies. We first list multiple standardized and publicly available CRC datasets from two imaging types: colonoscopy and histopathology. Secondly, we categorize the studies based on the different types of CRC detected (tumor tissue, microsatellite instability, and polyps), and we assess the data preprocessing steps and the adopted DL architectures before presenting the optimum diagnostic results. CRC diagnosis with DL algorithms is still in the preclinical phase, and therefore, we point out some open issues and provide some insights into the practicability and development of robust diagnostic systems in future health care and oncology.

Cerebellar and subcortical atrophy contribute to psychiatric symptoms in frontotemporal dementia

Recent studies have suggested that cerebellar and subcortical structures are impacted early in the disease progression of genetic frontotemporal dementia (FTD) due to microtubule-associated protein tau (MAPT), progranulin (GRN) and chromosome 9 open reading frame 72 (C9orf72). However, the clinical contribution of the structures involved in the cerebello-subcortical circuitry has been understudied in FTD given their potentially central role in cognition and behaviour processes. The present study aims to investigate whether there is an association between the atrophy of the cerebellar and subcortical structures, and neuropsychiatric symptoms (using the revised version of the Cambridge Behavioral Inventory, CBI-R) across genetic mutations and whether this association starts during the preclinical phase of the disease. Our study included 983 participants from the Genetic Frontotemporal dementia Initiative (GENFI) including mutation carriers (n=608) and non-carrier first-degree relatives of known symptomatic carriers (n= 375). Voxel-wise analysis of the thalamus, striatum, globus pallidus, amygdala, and the cerebellum was performed using deformation based morphometry (DBM) and partial least squares analyses (PLS) were used to link morphometry and behavioural symptoms. Our univariate results suggest that in this group of primarily presymptomatic subjects, volume loss in subcortical and cerebellar structure was primarily a function of aging, with only the C9orf72 group showing more pronounced volume loss in the thalamus compared to the non-carrier individuals. PLS analyses demonstrated that the cerebello-subcortical circuitry is related to all neuropsychiatric symptoms from the CBI-R, with significant overlap in brain/behaviour patterns, but also specificity for each genetic group. The biggest differences were in the extent of the cerebellar involvement (larger extent in C9orf72 group) and more prominent amygdalar contribution in the MAPT group. Finally, our findings demonstrated that C9orf72 and MAPT brain scores were related to estimated years before the age of symptom onset (EYO) in a second order relationship highlighting a steeper brain score decline 20 years before expected symptom onset, while GRN brain scores were related to age and not EYO. Overall, these results demonstrated the important role of the subcortical structures and especially of the cerebellum in genetic FTD symptom expression.

Radiotheranostic Agents Targeting Neuroblastoma: State-of-the-Art and Emerging Perspectives

Neuroblastoma (NB) represents the most common extracranial tumor of childhood. Prognosis is quite variable, ranging from spontaneous regression to aggressive behavior with wide metastatization, high mortality, and limited therapeutic options. Radiotheranostics combines a radiopharmaceutical pair in a unique approach, suitable both for diagnosis and therapy. For many years, metaiodobenzylguanidine (MIBG), labeled with 123I for imaging or 131I for therapy, has represented the main theranostic agent in NB, since up to 90% of NB incorporates the aforementioned radiopharmaceutical. In recent years, novel theranostic agents hold promise in moving the field of NB radiotheranostics forward. In particular, SarTATE, consisting of octreotate targeting somatostatin receptors, has been applied with encouraging results, with 64Cu-SARTATE being used for disease detection and with 67Cu-SARTATE being used for therapy. Furthermore, recent evidence has highlighted the potential of targeted alpha therapy (TAT) for treating cancer by virtue of alpha particles’ high ionizing density and high probability of killing cells along their track. On this path, 211At-astatobenzylguanidine (MABG) has been developed as a potential agent for TAT and is actually under evaluation in preclinical NB models. In this review, we performed a web-based and desktop literature research concerning radiotheranostic approaches in NB, covering both the radiopharmaceuticals already implemented in clinical practice (i.e.,123/1311-MIBG) and those still in a preliminary or preclinical phase.

Unveiling the Effect of Low pH on the SARS-CoV-2 Main Protease by Molecular Dynamics Simulations

(1) Background: Main Protease (Mpro) is an attractive therapeutic target that acts in the replication and transcription of the SARS-CoV-2 coronavirus. Mpro is rich in residues exposed to protonation/deprotonation changes which could affect its enzymatic function. This work aimed to explore the effect of the protonation/deprotonation states of Mpro at different pHs using computational techniques. (2) Methods: The different distribution charges were obtained in all the evaluated pHs by the Semi-Grand Canonical Monte Carlo (SGCMC) method. A set of Molecular Dynamics (MD) simulations was performed to consider the different protonation/deprotonation during 250 ns, verifying the structural stability of Mpro at different pHs. (3) Results: The present findings demonstrate that active site residues and residues that allow Mpro dimerisation was not affected by pH changes. However, Mpro substrate-binding residues were altered at low pHs, allowing the increased pocket volume. Additionally, the results of the solvent distribution around Sγ, Hγ, Nδ1 and Hδ1 atoms of the catalytic residues Cys145 and His41 showed a low and high-water affinity at acidic pH, respectively. It which could be crucial in the catalytic mechanism of SARS-CoV-2 Mpro at low pHs. Moreover, we analysed the docking interactions of PF-00835231 from Pfizer in the preclinical phase, which shows excellent affinity with the Mpro at different pHs. (4) Conclusion: Overall, these findings indicate that SARS-CoV-2 Mpro is highly stable at acidic pH conditions, and this inhibitor could have a desirable function at this condition.