Induction of epithelial-mesenchymal transition in thyroid follicular cells is associated with cell adhesion alterations and low-dose hyper-radiosensitivity.

BACKGROUND: Epithelial-mesenchymal transition (EMT) is associated with altered cellular adhesion. We previously demonstrated that cellular adhesion influences Low-dose Hyper-Radiosensitivity (HRS) in a variety of tumor cells. However, the relationship of low-dose HRS with the phenotypic plasticity incurred by EMT during the neoplastic transformation remains to be elucidated. OBJECTIVE: To investigate whether acquisition of EMT phenotype during progressive neoplastic transformation may affect low-dose radiation sensitivity. METHODS: Primary thyroid cells obtained from a human cystic thyroid nodule were first subjected to nutritional stress. This yielded immortalized INM-Thy1 cell strain, which was further treated with either multiple γ-radiation fractions (1.5 Gy each) or repetitive cycles of 3-methylcholanthrene and phorbol-12-myristate-13-acetate, yielding two progressive transformants, viz., INM-Thy1R and INM-Thy1C. Morphological alterations, chromosomal double-minutes, cell adhesion proteins, anchorage dependency, tumorigenicity in nude mice and cellular radiosensitivity were studied in these strains. RESULTS: Both transformants (INM-Thy1R, INM-Thy1C) displayed progressive tumorigenic features, viz., soft agar colony growth and solid tumor growth in nude mice, coupled with features of epithelial-mesenchymal transition and activated Wnt pathway. Incidentally, the chemical-induced transformant (INM-Thy1C) displayed a prominent HRS (αs/αr = 29.35) which remained unaffected at high cell density. However, the parental (INM-Thy1) cell line as well as radiation-induced transformant (INM-Thy1R) failed to show this hypersensitivity. CONCLUSION: The study shows that induction of EMT in thyroid follicular cells may accompany increased susceptibility to low-dose ionizing radiation, which was attenuated by adaptive resistance acquired during radiation-induced transformation.

Hemodynamic functional connectivity optimization of frequency EEG microstates enables attention LSTM framework to classify distinct temporal cortical communications of different cognitive tasks.

Temporal analysis of global cortical communication of cognitive tasks in coarse EEG information is still challenging due to the underlying complex neural mechanisms. This study proposes an attention-based time-series deep learning framework that processes fMRI functional connectivity optimized quasi-stable frequency microstates for classifying distinct temporal cortical communications of the cognitive task. Seventy volunteers were subjected to visual target detection tasks, and their electroencephalogram (EEG) and functional MRI (fMRI) were acquired simultaneously. At first, the acquired EEG information was preprocessed and bandpass to delta, theta, alpha, beta, and gamma bands and then subjected to quasi-stable frequency-microstate estimation. Subsequently, time-series elicitation of each frequency microstates is optimized with graph theory measures of simultaneously eliciting fMRI functional connectivity between frontal, parietal, and temporal cortices. The distinct neural mechanisms associated with each optimized frequency-microstate were analyzed using microstate-informed fMRI. Finally, these optimized, quasi-stable frequency microstates were employed to train and validate the attention-based Long Short-Term Memory (LSTM) time-series architecture for classifying distinct temporal cortical communications of the target from other cognitive tasks. The temporal, sliding input sampling windows were chosen between 180 to 750 ms/segment based on the stability of transition probabilities of the optimized microstates. The results revealed 12 distinct frequency microstates capable of deciphering target detections' temporal cortical communications from other task engagements. Particularly, fMRI functional connectivity measures of target engagement were observed significantly correlated with the right-diagonal delta (r = 0.31), anterior-posterior theta (r = 0.35), left-right theta (r = - 0.32), alpha (r = - 0.31) microstates. Further, neuro-vascular information of microstate-informed fMRI analysis revealed the association of delta/theta and alpha/beta microstates with cortical communications and local neural processing, respectively. The classification accuracies of the attention-based LSTM were higher than the traditional LSTM architectures, particularly the frameworks that sampled the EEG data with a temporal width of 300 ms/segment. In conclusion, the study demonstrates reliable temporal classifications of global cortical communication of distinct tasks using an attention-based LSTM utilizing fMRI functional connectivity optimized quasi-stable frequency microstates.

2-deoxy-D-glucose as an adjunct to standard of care in the medical management of COVID-19: a proof-of-concept and dose-ranging randomised phase II clinical trial.

BACKGROUND: At present, no single efficacious therapeutic exists for acute COVID-19 management and a multimodal approach may be necessary. 2-deoxy-D-glucose (2-DG) is a metabolic inhibitor that has been shown to limit multiplication of SARS-CoV-2 in-vitro. We evaluated the efficacy and safety of 2-DG as adjunct to standard care in the treatment of moderate to severe COVID-19 patients. METHODS: We conducted a randomized, open-label, phase II, clinical study to evaluate the efficacy, safety, and tolerability of 2-DG administered as adjunct to standard of care (SOC). A total of 110 patients between the ages of 18 and 65 years with moderate to severe COVID-19 were included. Patients were randomized to receive 63, 90, or 126 mg/kg/day 2-DG in addition to SOC or SOC only. Times to maintaining SpO2 ≥ 94% on room air, discharge, clinical recovery, vital signs normalisation, improvement by 1 and 2 points on WHO clinical progression scale, negative conversion on RT-PCR, requirement for intensive care, and mortality were analyzed to assess the efficacy. RESULTS: Patients treated with 90 mg/kg/day 2-DG plus SOC showed better outcomes. Time to maintaining SpO2 ≥ 94% was significantly shorter in the 2-DG 90 mg compared to SOC (median 2.5 days vs. 5 days, Hazard ratio [95% confidence interval] = 2.3 [1.14, 4.64], p = 0.0201). Times to discharge from isolation ward, to clinical recovery, and to vital signs normalization were significantly shorter for the 2-DG 90 mg group. All three doses of 2-DG were well tolerated. Thirty-three (30.3%) patients reported 65 adverse events and were mostly (86%) mild. CONCLUSIONS: 2-DG 90 mg/kg/day as adjunct to SOC showed clinical benefit over SOC alone in the treatment of moderate to severe COVID-19. The promising trends observed in current phase II study is encouraging for confirmatory evaluation of the efficacy and safety of 2-DG in a larger phase III trial. TRIAL REGISTRATION: CTRI, CTRI/2020/06/025664. Registered 5th June 2020, http://ctri.nic.in/Clinicaltrials/pdf_generate.php?trialid=44369&EncHid=&modid=&compid=%27,%2744369det%27 .

Urinary metabolic modulation in human participants residing in Siachen: a 1H NMR metabolomics approach.

The main physiological challenge in high altitude environment is hypoxia which affects the aerobic metabolism reducing the energy supply. These changes may further progress toward extreme environment-related diseases. These are further reflected in changes in small molecular weight metabolites and metabolic pathways. In the present study, metabolic changes due to chronic environmental hypoxia were assessed using 1H NMR metabolomics by analysing the urinary metabolic profile of 70 people at sea level and 40 people at Siachen camp (3700 m) for 1 year. Multivariate statistical analysis was carried out, and PLSDA detected 15 metabolites based on VIP score > 1. ROC analysis detected cis-aconitate, Nicotinamide Mononucleotide, Tyrosine, Choline and Creatinine metabolites with a high range of sensitivity and specificity. Pathway analysis revealed 16 pathways impact > 0.05, and phenylalanine tyrosine and tryptophan biosynthesis was the most prominent altered pathway indicating metabolic remodelling to meet the energy requirements. TCA cycle, Glycine serine and Threonine metabolism, Glutathione metabolism and Cysteine alterations were other metabolic pathways affected during long-term high-altitude hypoxia exposure. Present findings will help unlock a new dimension for the potential application of NMR metabolomics to address extreme environment-related health problems, early detection and developing strategies to combat high altitude hypoxia.

Glycolytic inhibitor 2-deoxy-d-glucose attenuates SARS-CoV-2 multiplication in host cells and weakens the infective potential of progeny virions.

AIMS: Virus-infected host cells switch their metabolism to a more glycolytic phenotype, required for new virion synthesis and packaging. Therefore, we investigated the effect and mechanistic action of glycolytic inhibitor 2-Deoxy-d-glucose (2-DG) on virus multiplication in host cells following SARS-CoV-2 infection. MAIN METHODS: SARS-CoV-2 induced change in glycolysis was examined in Vero E6 cells. Effect of 2-DG on virus multiplication was evaluated by RT-PCR (N and RdRp genes) analysis, protein expression analysis of Nucleocapsid (N) and Spike (S) proteins and visual indication of cytopathy effect (CPE), The mass spectrometry analysis was performed to examine the 2-DG induced change in glycosylation status of receptor binding domain (RBD) in SARS-CoV-2 spike protein. KEY FINDINGS: We observed SARS-COV-2 infection induced increased glucose influx and glycolysis, resulting in selectively high accumulation of the fluorescent glucose analog, 2-NBDG in Vero E6 cells. 2-DG inhibited glycolysis, reduced virus multiplication and alleviated cells from virus-induced cytopathic effect (CPE) in SARS-CoV-2 infected cells. The progeny virions produced from 2-DG treated cells were found unglycosylated at crucial N-glycosites (N331 and N343) of the receptor-binding domain (RBD) in the spike protein, resulting in production of defective progeny virions with compromised infective potential. SIGNIFICANCE: The mechanistic study revealed that the inhibition of SARS-COV-2 multiplication is attributed to 2-DG induced glycolysis inhibition and possibly un-glycosylation of the spike protein, also. Therefore, based on its previous human trials in different types of Cancer and Herpes patients, it could be a potential molecule to study in COVID-19 patients.

Functional connectivity between frontal/parietal regions and MTL-basal ganglia during feedback learning and declarative memory retrieval.

Feedback assists the memory system in preserving the learnings from ongoing activities and updating it for future retrievals. Thus, the feedback coming from an individual's performance affects the behavior and, thereby, the performance. However, little is known regarding the interactions of learning and memory associated regions. Thus, we employ a combination of functional connectivity and neurovascular approach to explore the significance of these interactions. Our study comprises thirty-five volunteers who undergo a feedback declarative memory task using simultaneous EEG-fMRI data acquisition. Functional connectivity analysis showed that medial temporal lobe (MTL) and basal ganglia possess significant connectivity but differential relationships during feedback learning and memory retrieval. Specifically, Putamen and pallidum (sub-regions of basal ganglia) are the central hubs in these mechanisms. The neurovascular analysis reveals the increased correlation of frontal-alpha and theta powers with the bold activity of MTL during memory retrievals. The results also report the role of the frontal (and parietal) alpha-beta powers in de-synchronization (and synchronization) of the bold activity of caudate; and parietal-theta (frontal-higher-alpha) power in de-synchronization (and synchronization) of bold activity of right accumbens. Hence, the study demonstrates the significant role of the frontal-parietal EEG powers in MTL-basal ganglia relationships and neuronal adaptations during declarative memory retrieval.

Assessment of distinct subcortical and cortical contributions to affect and approach/withdrawal behavior by means of resting-state functional connectivity approach.

The relevance of subcortical structures and interhemispheric subcortical-cortical interactions among positive/negative affect and approach/withdrawal tendencies during resting-state are not fully understood. Gaining this knowledge may foster the know-how on their role in subsequent task-engagement and also on the interlinkage among affective measures and approach/withdrawal dichotomy. Here we performed a study based on Region of Interest (ROI)-based analysis and graph-theory estimates for global and subnetworks on a limited sample of healthy 50 male volunteers who recorded resting functional magnetic resonance imaging (fMRI) and self-reported measures of Positive and Negative Affect Schedule (PANAS) and BAS-BIS (Behavioral Activation System-Behavioral Inhibition System) scores. Our study's initial results of region of interest-to-region of interest (ROI-to-ROI) connectivity revealed the connectivity of subcortical neural substrates of PANAS and BAS-BIS scores with bilateralized cortical regions. However, on probing the lateralization of strength of degree measures of the cortical-regions vital for subcortical-cortical interaction, we found, for positive affect, a left-hemispheric proclivity. Further, higher connectivity within the left hemisphere was also observed for the left-lateralized critical cortical regions of negative affect. Our study also revealed the association of emotion and memory-related subcortical-cortical interactions in positive and negative affect. Right amygdala-right thalamus-frontotemporal cortical areas emanated in positive affect, and right putamen-left hippocampus-frontotemporal cortical regions network stemmed for negative affect. Then, we show the involvement of basal-ganglia structures in approach-withdrawal dichotomy with tight coupling of right-caudate, left-accumbens with anterior cingulate, and insular regions for withdrawal/inhibition system. Further, distinct involvement of the insula (posterior) in affective states while insula (anterior) in approach/withdrawal systems builds up for the existence of a feedback loop between affective and approach/withdrawal systems. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Microstates-based resting frontal alpha asymmetry approach for understanding affect and approach/withdrawal behavior.

The role of resting frontal alpha-asymmetry in explaining neural-mechanisms of affect and approach/withdrawal behavior is still debatable. The present study explores the ability of the quasi-stable resting EEG asymmetry information and the associated neurovascular synchronization/desynchronization in bringing more insight into the understanding of neural-mechanisms of affect and approach/withdrawal behavior. For this purpose, a novel frontal alpha-asymmetry based on microstates, that assess quasi-stable EEG scalp topography information, is proposed and compared against standard frontal-asymmetry. Both proposed and standard frontal alpha-asymmetries were estimated from thirty-nine healthy volunteers resting-EEG simultaneously acquired with resting-fMRI. Further, neurovascular mechanisms of these asymmetry measures were estimated through EEG-informed fMRI. Subsequently, the Hemodynamic Lateralization Index (HLI) of the neural-underpinnings of both asymmetry measures was assessed. Finally, the robust correlation of both asymmetry-measures and their HLI's with PANAS, BIS/BAS was carried out. The standard resting frontal-asymmetry and its HLI yielded no significant correlation with any psychological-measures. However, the microstate resting frontal-asymmetry correlated significantly with negative affect and its neural underpinning's HLI significantly correlated with Positive/Negative affect and BIS/BAS measures. Finally, alpha-BOLD desynchronization was observed in neural-underpinning whose HLI correlated significantly with negative affect and BIS. Hence, the proposed resting microstate-frontal asymmetry better assesses the neural-mechanisms of affect, approach/withdrawal behavior.

Effects of Neural Mechanisms of Pretask Resting EEG Alpha Information on Situational Awareness: A Functional Connectivity Approach.

OBJECTIVE: In this study, the influence of pretask resting neural mechanisms on situational awareness (SA)-task is studied. BACKGROUND: Pretask electroencephalography (EEG) information and Stroop effect are known to influence task engagement independently. However, neural mechanisms of pretask resting absolute alpha (PRAA) and pretask resting alpha frontal asymmetry (PRAFA) in influencing SA-task which is undergoing Stroop effect is still not understood. METHOD: The study involved pretask resting EEG measurements from 18 healthy individuals followed by functional magnetic resonance imaging (fMRI) acquisition during SA-task. To understand the effect of pretask alpha information and Stroop effect on SA, a robust correlation between mean reaction time, SA Index, PRAA, and PRAFA were assessed. Furthermore, neural underpinnings of PRAA, PRAFA in SA-task, and functional connectivity were analyzed through the EEG-informed fMRI approach. RESULTS: Significant robust correlation of reaction time was observed with SA Index (Pearson: r = .50, pcorr = .05) and PRAFA (Pearson: r = .63; pcorr = .01), respectively. Similarly, SA Index significantly correlated with PRAFA (Pearson: r = .56, pcorr = .01; Spearman: r = .61, pcorr = .007), and PRAA (Pearson: r = .59, pcorr = .005; Spearman: r = .59, pcorr = .002). Neural underpinnings of SA-task revealed regions involved in visual-processing and higher-order cognition. PRAA was primarily underpinned at frontal-temporal areas and functionally connected to SA-task regions pertaining to the emotional regulation. PRAFA has correlated with limbic and parietal regions, which are involved in integration of visual, emotion, and memory information of SA-task. CONCLUSION: The results suggest a strong association of reaction time with SA-task and PRAFA and strongly support the hypothesis that PRAFA, PRAA, and associated neural mechanisms significantly influence the outcome of SA-task. APPLICATION: It is beneficial to study the effect of pretask resting information on SA-task to improve SA.

Estimation of Functional Connectivity Modulations During Task Engagement and Their Neurovascular Underpinnings Through Hemodynamic Reorganization Method.

This study proposes an approach to understand the effect of task engagement through integrated analysis of modulations in functional networks and associated changes in their neurovascular underpinnings at every voxel. For this purpose, a novel approach that brings reorganization in acquired task-functional magnetic resonance imaging information based on hemodynamic characteristics of every task stimulus is proposed and validated. At first, modulations in functional networks of visual target detection task were estimated at every voxel through proposed methodology. It revealed task stimulus dependency in the modulation of default mode network (DMN). The DMN modulated as task negative network (TNN) during target stimulus. On the contrary, it was not entirely TNN during nontarget stimulus. The frontal-parietal and visual networks modulated as task positive network during both task stimuli. Further, modulations of neurovascular underpinnings associated with engagement of task were estimated by correlating the hemodynamically reorganized task blood oxygen level dependent information with simultaneously acquired electroencephalography frequency powers. It revealed the strong association of neurovascular underpinnings with their modulation of functional networks and the associated neuronal activity during task engagement. Finally, graph theoretical parameters such as local, global efficiency and clustering coefficient were also measured at the specific regions for validating the results of proposed method. Modulation observed in graph theory measures clearly validated the activation and deactivation of functional networks observed by the proposed method during task engagement. Thus, the voxel-wise estimation of task-related modulation of functional networks and associated neurovascular underpinnings through proposed technique provide better insights into neuronal mechanism involved during engagement in a task.

Assessment of cochlear nerve deficiency and its effect on normal maturation of auditory tract by diffusion kurtosis imaging and diffusion tensor imaging: A correlational approach.

OBJECTIVE: The use of diffusion kurtosis imaging (DKI) and diffusion tensor imaging (DTI) was evaluated in assessing cochlear nerve deficiency (CND) and its effect on normal maturation of auditory tract. METHODS: 25 CND patients and 25 controls (age matched: 2months to 17years, gender matched) were evaluated by mean kurtosis (MK), axial kurtosis, and radial kurtosis and compared against fractional anisotropy (FA), axial and radial diffusivities in internal auditory canal (IAC), lateral lemniscus (LL) and inferior colliculus (IC). The age related changes of auditory tract were studied through Pearson correlation between estimated indices and age of both CND and control populations. RESULTS: Significant loss of MK (IAC: 10.71%, IC: 10.87%, LL: 15.63%) was observed in CND cases as against moderate reduction in FA (IAC: 8.57%, IC: 10%, LL: 7.69%) in all three anatomical locations. Similarly, substantial decline is observed in radial kurtosis (IAC: 27.03%, IC: 33.33%, LL: 31.43%) in comparison to moderate increase in radial diffusivity (IAC: 13.46%, IC: 24.39%, LL: 24%) in CND cases. No statistically significant change was seen in both axial kurtosis and diffusivities. In control populations, MK (r=0.473, p=0.011) and radial kurtosis (r=0.418, p=0.016) correlate positively with age and had no correlation in case of CND cases. FA (r=0.356, p=0.019) minimally correlated with age in control population but showed no statistically significant correlation in CND cases (r=0.198, p=0.036). CONCLUSION: DKI metrics performed better than DTI in assessing microstructural changes of CND. In particular, MK and radial kurtosis are found to be more sensitive enough to differentiate the normal maturation of cochlear nerve from CND cases.