bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Flow Cytometric Immunoassay for Detection of Proinsulin Autoantibodies in Diabetes Mellitus Employing a Recombinant Autoantigen Expressed in E. coli
Authors: Sabljic Adriana Victoria , Bombicino Silvina Sonia , Marfía Juan Ignacio , Guerra Luciano Lucas , Steinhardt Alberto Penas , Faccinetti Natalia Inés , Iacono Rubén Francisco , Poskus Edgardo , Trabucchi Aldana , Valdez Silvina Noemí
Published: April 2021

Introduction

Insulin and proinsulin autoantibodies (IAA/PAA) are usually the first markers to appear in patients with type 1 Diabetes Mellitus (T1DM) and their prevalence ranges from 10 to 60% in the child-adolescent population. The reference method for IAA/PAA detection is the Radioligand Binding Assay (RBA), a highly specific and sensitive technique, but expensive and polluting. The aim of this work was to develop a novel flow cytometric microsphere-based immunoassay (FloCMIA) for PAA detection, employing recombinant human proinsulin (PI), as an alternative method to RBA, less expensive and harmful to the environment.

Materials and Methods

Human PI was expressed as Thioredoxin fusion protein (TrxPI) in E. coli and a fraction was biotinylated. A double paratope model was used in which samples were incubated with TrxPI–biotin and microspheres adsorbed with TrxPI. The immune complexes were revealed using Streptavidin–Phycoerythrin. The geometric mean of the signals was analyzed, and the results were expressed as Standard Deviation scores (SDs). Sera from 100 normal human control and from 111 type 1 diabetic patients were evaluated by FloCMIA. To correlate the novel assay with RBA, 51 diabetic patients were selected, spanning a wide range of PAA reactivity by RBA.

Results

The study of ROC curves allowed choosing a cut-off value of 3.0 SDs and the AUC was 0.705, indicating that FloCMIA has fair ability to distinguish between samples from each group. A prevalence of 50% for PAA was obtained in the population of diabetic patients studied. The specificity was 96% and the analytical sensitivity (percentage of patients RBA positive, also positive by FloCMIA) was 69%. There was a substantial agreement between methods (kappa statistic=0.700).

Conclusions

A novel immunoassay based on flow cytometry that uses easy-to produce recombinant PI was developed. This assay constitutes an innovative and cost-effective alternative to RBA for the determination of PAA in patients’ sera. The method developed here, presents good performance and a wide dynamic range together with a small required sample volume. Furthermore, these results make it possible to develop multiplex immunoassays that allow the combined detection of autoantibodies present in T1DM and other related autoimmune diseases.
EVA1B to Evaluate the Tumor Immune Microenvironment and Clinical Prognosis in Glioma
Authors: Qu Shanqiang , Liu Jin , Wang Huafu
Published: April 2021

Background

Previous research indicated that the tumor cells and microenvironment interactions are critical for the immunotherapeutic response. However, predicting the clinical response to immunotherapy remains a dilemma for clinicians. Hence, this study aimed to investigate the associations between EVA1B expression and prognosis and tumor-infiltrating immune cells in glioma.

Methods

Firstly, we detected the EVA1B expression in glioma tissues through biological databases. The chi-squared test, Kaplan-Meier, and univariate and multivariate Cox regression analyses were used to analyze the clinical significance of EVA1B expression. The correlation between EVA1B expression and levels of tumor-infiltrating immune cells in glioma tissues was investigated. Receiver operating characteristic (ROC) analysis was performed to compare the predictive power between EVA1B and other commonly immune-related markers.

Results

In the CGGA cohort of 325 glioma patients, we found that EVA1B was upregulated in glioma, and increased with tumor grade. High EVA1B expression was prominently associated with unfavorable clinicopathological features, and poorer survival of patients, which were further confirmed by TCGA (n=609) and GEO (n=74) cohorts. Furthermore, multivariate analysis indicated that EVA1B is an independent prognostic biomarker for glioma. Importantly, EVA1B overexpression was associated with a higher infiltration level of CD4+ T cells, CD8+ T cells, B cells, macrophages, and neutrophils in glioma. ROC curves showed that, compared with PD-L1, CTLA-4, and Siglec15, EVA1B presented a higher area under the curve (AUC) value (AUC=0.824) for predicting high immune infiltration levels in glioma.

Conclusions

We found that EVA1B was upregulated and could act as a poor prognostic biomarker in glioma. Importantly, EVA1B overexpression was associated with the immune infiltration levels of immune cells including B cells, CD4+ T cells, CD8+ T cells, macrophages, and neutrophils, and strongly with the overall immune infiltration levels of glioma. These findings suggested that EVA1B might be a potential biomarker for evaluating prognosis and immune infiltration in glioma.
Clonally Focused Public and Private T Cells in Resected Brain Tissue From Surgeries to Treat Children With Intractable Seizures
Authors: Chang Julia W. , Reyes Samuel D. , Faure-Kumar Emmanuelle , Lam Sandi K. , Lawlor Michael W. , Leventer Richard J. , Lew Sean M. , Lockhart Paul J. , Pope Kathryn , Weiner Howard L. , Salamon Noriko , Vinters Harry V. , Mathern Gary W. , Fallah Aria , Owens Geoffrey C.
Published: April 2021

Using a targeted transcriptomics approach, we have analyzed resected brain tissue from a cohort of 53 pediatric epilepsy surgery cases, and have found that there is a spectrum of involvement of both the innate and adaptive immune systems as evidenced by the differential expression of immune-specific genes in the affected brain tissue. The specimens with the highest expression of immune-specific genes were from two Rasmussen encephalitis cases, which is known to be a neuro-immunological disease, but also from tuberous sclerosis complex (TSC), focal cortical dysplasia, and hemimegalencephaly surgery cases. We obtained T cell receptor (TCR) Vβ chain sequence data from brain tissue and blood from patients with the highest levels of T cell transcripts. The clonality indices and the frequency of the top 50 Vβ clonotypes indicated that T cells in the brain were clonally restricted. The top 50 Vβ clonotypes comprised both public and private (patient specific) clonotypes, and the TCR Vβ chain third complementarity region (CDR3) of the most abundant public Vβ clonotype in each brain sample was strikingly similar to a CDR3 that recognizes an immunodominant epitope in either human cytomegalovirus or Epstein Barr virus, or influenza virus A. We found that the frequency of 14 of the top 50 brain Vβ clonotypes from a TSC surgery case had significantly increased in brain tissue removed to control recurrent seizures 11 months after the first surgery. Conversely, we found that the frequency in the blood of 18 of the top 50 brain clonotypes from a second TSC patient, who was seizure free, had significantly decreased 5 months after surgery indicating that T cell clones found in the brain had contracted in the periphery after removal of the brain area associated with seizure activity and inflammation. However, the frequency of a public and a private clonotype significantly increased in the brain after seizures recurred and the patient underwent a second surgery. Combined single cell gene expression and TCR sequencing of brain-infiltrating leukocytes from the second surgery showed that the two clones were CD8 effector T cells, indicating that they are likely to be pathologically relevant.
Assessing the Skill of a High-Resolution Marine Biophysical Model Using Geostatistical Analysis of Mesoscale Ocean Chlorophyll Variability From Field Observations and Remote Sensing
Authors: Eveleth Rachel , Glover David M. , Long Matthew C. , Lima Ivan D. , Chase Alison P. , Doney Scott C.
Published: April 2021

High-resolution ocean biophysical models are now routinely being conducted at basin and global-scale, opening opportunities to deepen our understanding of the mechanistic coupling of physical and biological processes at the mesoscale. Prior to using these models to test scientific questions, we need to assess their skill. While progress has been made in validating the mean field, little work has been done to evaluate skill of the simulated mesoscale variability. Here we use geostatistical 2-D variograms to quantify the magnitude and spatial scale of chlorophyll a patchiness in a 1/10th-degree eddy-resolving coupled Community Earth System Model simulation. We compare results from satellite remote sensing and ship underway observations in the North Atlantic Ocean, where there is a large seasonal phytoplankton bloom. The coefficients of variation, i.e., the arithmetic standard deviation divided by the mean, from the two observational data sets are approximately invariant across a large range of mean chlorophyllavalues from oligotrophic and winter to subpolar bloom conditions. This relationship between the chlorophyllamesoscale variability and the mean field appears to reflect an emergent property of marine biophysics, and the high-resolution simulation does poorly in capturing this skill metric, with the model underestimating observed variability under low chlorophyllaconditions such as in the subtropics.
Challenges of Gauging the Impact of Area-Based Fishery Closures and OECMs: A Case Study Using Long-Standing Canadian Groundfish Closures
Authors: Shackell Nancy L. , Keith David M. , Lotze Heike K.
Published: April 2021

The United Nations Convention on Biological Diversity was established in 1993. Canada is a signatory nation that has adopted, and exceeded, the UN Aichi biodiversity target to protect 10% of coastal and marine areas through marine protected areas or “other effective area-based conservation measures” (OECMs) by 2020. However, the science of OECMs as contributors to biodiversity conservation is relatively young and their definition and efficacy testing continue to evolve. Here, we examine whether areas closed to fishing on the Scotian Shelf in Atlantic Canada, where the groundfish community had collapsed in the early 1990s, have the potential to serve as OECMs for groundfish recovery. Using long-term research survey data, we show that three long-term area-based fishing fleet closures did not enhance per capita population growth rates of the majority of 24 common groundfish species. At a regional scale, 10 out of 24 species are currently at less than 50% of their pre-collapse (1979–1992) biomass, reflecting a sustained diminished productivity, even though fishing mortality has been drastically reduced through a moratorium in 1993. Additional measures are needed to protect severely depleted groundfish, especially when the causes of continued diminished productivity are still largely unresolved. The importance of OECMs as a risk-averse approach toward sustainability is globally accepted and they can be considered a tool toward the overarching UN Sustainable Development Goals (SDG-14). Our study provides further impetus toward articulating the criteria of OECMs and improving their design, monitoring, and testing, while placing OECMs within the broader context of sustainable ecosystem-based management.
Ross Sea Benthic Ecosystems: Macro- and Mega-faunal Community Patterns From a Multi-environment Survey
Authors: Cummings Vonda J. , Bowden David A. , Pinkerton Matthew H. , Halliday N. Jane , Hewitt Judi E.
Published: April 2021

The Ross Sea, Antarctica, is amongst the least human-impacted marine environments, and the site of the world’s largest Marine Protected Area. We present research on two components of the Ross Sea benthic fauna: mega-epifauna, and macro-infauna, sampled using video and multicore, respectively, on the continental shelf and in previously unsampled habitats on the northern continental slope and abyssal plain. We describe physical habitat characteristics and community composition, in terms of faunal diversity, abundance, and functional traits, and compare similarities within and between habitats. We also examine relationships between faunal distributions and ice cover and productivity, using summaries of satellite-derived data over the decade prior to our sampling. Clear differences in seafloor characteristics and communities were noted between environments. Seafloor substrates were more diverse on the Slope and Abyss, while taxa were generally more diverse on the Shelf. Mega-epifauna were predominantly suspension feeders across the Shelf and Slope, with deposit feeder-grazers found in higher or equal abundances in the Abyss. In contrast, suspension feeders were the least common macro-infaunal feeding type on the Shelf and Slope. Concordance between the mega-epifauna and macro-infauna data suggests that non-destructive video sampling of mega-epifauna can be used to indicate likely composition of macro-infauna, at larger spatial scales, at least. Primary productivity, seabed organic flux, and sea ice concentrations, and their variability over time, were important structuring factors for both community types. This illustrates the importance of better understanding bentho-pelagic coupling and incorporating this in biogeographic and process-distribution models, to enable meaningful predictions of how these ecosystems may be impacted by projected environmental changes. This study has enhanced our understanding of the distributions and functions of seabed habitats and fauna inside and outside the Ross Sea MPA boundaries, expanding the baseline dataset against which the success of the MPA, as well as variability and change in benthic communities can be evaluated longer term.
Development and Validation of Risk Prediction Model for In-hospital Mortality Among Patients Hospitalized With Acute Exacerbation Chronic Obstructive Pulmonary Disease Between 2015 and 2019
Authors: Dong Fen , Ren Xiaoxia , Huang Ke , Wang Yanyan , Jiao Jianjun , Yang Ting
Published: April 2021

Background: In patients with chronic obstructive pulmonary disease (COPD), acute exacerbations affect patients' health and can lead to death. This study was aimed to develop a prediction model for in-hospital mortality in patients with acute exacerbations of COPD (AECOPD).

Method: A retrospective study was performed in patients hospitalized for AECOPD between 2015 and 2019. Patients admitted between 2015 and 2017 were included to develop model and individuals admitted in the following 2 years were included for external validation. We analyzed variables that were readily available in clinical practice. Given that death was a rare outcome in this study, we fitted Firth penalized logistic regression. C statistic and calibration plot quantified the model performance. Optimism-corrected C statistic and slope were estimated by bootstrapping. Accordingly, the prediction model was adjusted and then transformed into risk score.

Result: Between 2015 and 2017, 1,096 eligible patients were analyzed, with a mean age of 73 years and 67.8% male. The in-hospital mortality was 2.6%. Compared to survivors, non-survivors were older, more admitted from emergency, more frequently concomitant with respiratory failure, pneumothorax, hypoxic-hypercarbic encephalopathy, and had longer length of stay (LOS). Four variables were included into the final model: age, respiratory failure, pneumothorax, and LOS. In internal validation, C statistic was 0.9147, and the calibration slope was 1.0254. Their optimism-corrected values were 0.90887 and 0.9282, respectively, indicating satisfactory discrimination and calibration. When externally validated in 700 AECOPD patients during 2018 and 2019, the model demonstrated good discrimination with a C statistic of 0.8176. Calibration plot illustrated a varying discordance between predicted and observed mortality. It demonstrated good calibration in low-risk patients with predicted mortality rate ≤10% (P = 0.3253) but overestimated mortality in patients with predicted rate >10% (P < 0.0001). The risk score of 20 was regarded as a threshold with an optimal Youden index of 0.7154.

Conclusion: A simple prediction model for AECOPD in-hospital mortality has been developed and externally validated. Based on available data in clinical setting, the model could serve as an easily used instrument for clinical decision-making. Complications emerged as strong predictors, underscoring an important role of disease management in improving patients' prognoses during exacerbation episodes.
Susceptibility of ApoB and PCSK9 Genetic Polymorphisms to Diabetic Kidney Disease Among Chinese Diabetic Patients
Authors: Ma Liang , Wang Shaoting , Zhao Hailing , Yu Meijie , Deng Xiangling , Jiang Yongwei , Cao Yongtong , Li Ping , Niu Wenquan
Published: April 2021

This study aimed to investigate the susceptibility of 8 polymorphisms in ApoB and PCSK9 genes to diabetic kidney disease (DKD) in Chinese patients with type 2 diabetes mellitus. This is a case-control association study, including 575 DKD cases and 653 controls. Genotypes were determined using ligase detection reaction method, and data are analyzed using STATA software. The genotype distributions of rs1042034 and rs12720838 differed significantly between the two groups (P < 0.001 and P = 0.008, respectively). After adjusting for confounding factors, the mutations of rs1042034 and rs12720838 were associated with the significantly increased risk of DKD. For instance, carriers of rs1042034 T allele (CT and TT genotypes) were 1.07 times more likely to have DKD than carriers of rs1042034 CC genotype [odds ratio (OR) = 1.07, 95% confidence interval (CI): 1.03–1.10, P < 0.001]. Further, haplotype T-A-G-T in ApoB gene was overrepresented in cases (18.10%) compared with controls (12.76%) (PSimulated = 0.045), and haplotype T-A-G-T was associated with a 33% increased risk of DKD (OR = 1.33, 95% CI: 1.04, 1.70). In further haplotype-phenotype analysis, significant association was only noted for hypertension and omnibus haplotypes in ApoB gene (PSimulated = 0.001). Our findings indicate that ApoB gene is a candidate gene for DKD in Chinese patients with type 2 diabetes mellitus.
Mosquito Trilogy: Microbiota, Immunity and Pathogens, and Their Implications for the Control of Disease Transmission
Authors: Gabrieli Paolo , Caccia Silvia , Varotto-Boccazzi Ilaria , Arnoldi Irene , Barbieri Giulia , Comandatore Francesco , Epis Sara
Published: April 2021

In mosquitoes, the interaction between the gut microbiota, the immune system, and the pathogens that these insects transmit to humans and animals is regarded as a key component toward the development of control strategies, aimed at reducing the burden of severe diseases, such as malaria and dengue fever. Indeed, different microorganisms from the mosquito microbiota have been investigated for their ability to affect important traits of the biology of the host insect, related with its survival, development and reproduction. Furthermore, some microorganisms have been shown to modulate the immune response of mosquito females, significantly shaping their vector competence. Here, we will review current knowledge in this field, focusing on i) the complex interaction between the intestinal microbiota and mosquito females defenses, both in the gut and at humoral level; ii) how knowledge on these issues contributes to the development of novel and targeted strategies for the control of mosquito-borne diseases such as the use of paratransgenesis or taking advantage of the relationship between Wolbachia and mosquito hosts. We conclude by providing a brief overview of available knowledge on microbiota-immune system interplay in major insect vectors.
Influence of Non-canonical DNA Bases on the Genomic Diversity of Tevenvirinae
Authors: Nikulin Nikita A. , Zimin Andrei A.
Published: April 2021

TheTevenvirinaeviruses are some of the most common viruses on Earth. Representatives of this subfamily have long been used in the molecular biology studies as model organisms – since the emergence of the discipline.Tevenvirinaeare promising agents for phage therapy in animals and humans, since their representatives have only lytic life cycle and many of their host bacteria are pathogens. As confirmed experimentally, someTevenvirinaehave non-canonical DNA bases. Non-canonical bases can play an essential role in the diversification of closely related viruses. The article performs a comparative and evolutionary analysis ofTevenvirinaegenomes and components ofTevenvirinaegenomes. A comparative analysis of these genomes and the genes associated with the synthesis of non-canonical bases allows us to conclude that non-canonical bases have a major influence on the divergence ofTevenvirinaeviruses within the same habitats. Supposedly,Tevenvirinaedeveloped a strategy for changing HGT frequency in individual populations, which was based on the accumulation of proteins for the synthesis of non-canonical bases and proteins that used those bases as substrates. Owing to this strategy, ancestors ofTevenvirinaewith the highest frequency of HGT acquired genes that allowed them to exist in a certain niche, and ancestors with the lowest HGT frequency preserved the most adaptive of those genes. Given the origin and characteristics of genes associated with the synthesis of non-canonical bases inTevenvirinae, one can assume that other phages may have similar strategies. The article demonstrates the dependence of genomic diversity of closely relatedTevenvirinaeon non-canonical bases.
Phosphoproteome Study of Escherichia coli Devoid of Ser/Thr Kinase YeaG During the Metabolic Shift From Glucose to Malate
Authors: Sultan Abida , Jers Carsten , Ganief Tariq A. , Shi Lei , Senissar Meriem , Køhler Julie Bonne , Macek Boris , Mijakovic Ivan
Published: April 2021

Understanding phosphorylation-mediated regulation of metabolic enzymes, pathways, and cell phenotypes under metabolic shifts represents a major challenge. The kinases associated with most phosphorylation sites and the link between phosphorylation and enzyme activity remain unknown. In this study, we performed stable isotope labeling by amino acids in cell culture (SILAC)-based proteome and phosphoproteome analysis of Escherichia coli ΔyeaG, a strain lacking a poorly characterized serine/threonine kinase YeaG, to decipher kinase-substrate interactions and the effects on metabolic phenotype during shifts from glucose to malate. The starting point of our analysis was the identification of physiological conditions under which ΔyeaG exhibits a clear phenotype. By metabolic profiling, we discovered that ΔyeaG strain has a significantly shorter lag phase than the wild type during metabolic shift from glucose to malate. Under those conditions, our SILAC analysis revealed several proteins that were differentially phosphorylated in the ΔyeaG strain. By focusing on metabolic enzymes potentially involved in central carbon metabolism, we narrowed down our search for putative YeaG substrates and identified isocitrate lyase AceA as the direct substrate of YeaG. YeaG was capable of phosphorylating AceA in vitro only in the presence of malate, suggesting that this phosphorylation event is indeed relevant for glucose to malate shift. There is currently not enough evidence to firmly establish the exact mechanism of this newly observed regulatory phenomenon. However, our study clearly exemplifies the usefulness of SILAC-based approaches in identifying proteins kinase substrates, when applied in physiological conditions relevant for the activity of the protein kinase in question.
The Emerging Role of the Interplay Among Astrocytes, Microglia, and Neurons in the Hippocampus in Health and Disease
Authors: Lana Daniele , Ugolini Filippo , Nosi Daniele , Wenk Gary L. , Giovannini Maria Grazia
Published: April 2021

For over a century, neurons have been considered the basic functional units of the brain while glia only elements of support. Activation of glia has been long regarded detrimental for survival of neurons but more it appears that this is not the case in all circumstances. In this review, we report and discuss the recent literature on the alterations of astrocytes and microglia during inflammaging, the low-grade, slow, chronic inflammatory response that characterizes normal brain aging, and in acute inflammation. Becoming reactive, astrocytes and microglia undergo transcriptional, functional, and morphological changes that transform them into cells with different properties and functions, such as A1 and A2 astrocytes, and M1 and M2 microglia. This classification of microglia and astrocytes in two different, all-or-none states seems too simplistic, and does not correspond to the diverse variety of phenotypes so far found in the brain. Different interactions occur among the many cell populations of the central nervous system in health and disease conditions. Such interactions give rise to networks of morphological and functional reciprocal reliance and dependency. Alterations affecting one cell population reverberate to the others, favoring or dysregulating their activities. In the last part of this review, we present the modifications of the interplay between neurons and glia in rat models of brain aging and acute inflammation, focusing on the differences between CA1 and CA3 areas of the hippocampus, one of the brain regions most susceptible to different insults. With triple labeling fluorescent immunohistochemistry and confocal microscopy (TIC), it is possible to evaluate and compare quantitatively the morphological and functional alterations of the components of the neuron-astrocyte-microglia triad. In the contiguous and interconnected regions of rat hippocampus, CA1 and CA3 Stratum Radiatum, astrocytes and microglia show a different, finely regulated, and region-specific reactivity, demonstrating that glia responses vary in a significant manner from area to area. It will be of great interest to verify whether these differential reactivities of glia explain the diverse vulnerability of the hippocampal areas to aging or to different damaging insults, and particularly the higher sensitivity of CA1 pyramidal neurons to inflammatory stimuli.
P2X7 Receptors in Neurodegeneration: Potential Therapeutic Applications From Basic to Clinical Approaches
Authors: Territo Paul R. , Zarrinmayeh Hamideh
Published: April 2021

Purinergic receptors play important roles in central nervous system (CNS), where the bulk of these receptors are implicated in neuroinflammatory responses and regulation of cellular function of neurons, microglial and astrocytes. Within the P2X receptor family, P2X7receptor is generally known for its inactivity in normal conditions and activation by moderately high concentrations (>100 μM) of extracellular adenosine 5′-triphosphate (ATP) released from injured cells as a result of brain injury or pathological conditions. Activation of P2X7R contributes to the activation and proliferation of microglia and directly contribute to neurodegeneration by provoking microglia-mediated neuronal death, glutamate-mediated excitotoxicity, and NLRP3 inflammasome activation that results in initiation, maturity and release of the pro-inflammatory cytokines and generation of reactive oxygen and nitrogen species. These components of the inflammatory response play important roles in many neural pathologies and neurodegeneration disorders. In CNS, expression of P2X7R on microglia, astrocytes, and oligodendrocytes are upregulated under neuroinflammatory conditions. Severalin vivostudies have demonstrated beneficial effects of the P2X7receptor antagonists in animal model systems of neurodegenerative diseases. A number of specific and selective P2X7receptor antagonists have been developed, but only few of them have shown efficient brain permeability. Finding potent and selective P2X7receptor inhibitors which are also CNS penetrable and display acceptable pharmacokinetics (PK) has presented challenges for both academic researchers and pharmaceutical companies. In this review, we discuss the role of P2X7receptor function in neurodegenerative diseases, the pharmacological inhibition of the receptor, and PET radiopharmaceuticals which permit non-invasive monitoring of the P2X7receptor contribution to neuroinflammation associated with neurodegeneration.
Serotonergic Modulation of Persistent Inward Currents in Serotonergic Neurons of Medulla in ePet-EYFP Mice
Authors: Cheng Yi , Song Nan , Ge Renkai , Dai Yue
Published: April 2021

Serotonergic (5-HT) neurons in the medulla play multiple functional roles associated with many symptoms and motor activities. The descending serotonergic pathway from medulla is essential for initiating locomotion. However, the ionic properties of 5-HT neurons in the medulla remain unclear. Using whole-cell patch-clamp technique, we studied the biophysical and modulatory properties of persistent inward currents (PICs) in 5-HT neurons of medulla in ePet-EYFP transgenic mice (P3–P6). PICs were recorded by a family of voltage bi-ramps (10-s duration, 40-mV peak step), and the ascending and descending PICs were mirrored to analyze the PIC hysteresis. PICs were found in 77% of 5-HT neurons (198/258) with no significant difference between parapyramidal region (n = 107) and midline raphe nuclei (MRN) (n = 91) in either PIC onset (−47.4 ± 10 mV and −48.7 ± 7 mV; P = 0.44) or PIC amplitude (226.9 ± 138 pA and 259.2 ± 141 pA; P = 0.29). Ninety-six percentage (191/198) of the 5-HT neurons displayed counterclockwise hysteresis and four percentage (7/198) exhibited the clockwise hysteresis. The composite PICs could be differentiated as calcium component (Ca_PIC) by bath application of nimodipine (25 μM), sodium component (Na_PIC) by tetrodotoxin (TTX, 2 μM), and TTX- and dihydropyridine-resistance component (TDR_PIC) by TTX and nimodipine. Ca_PIC, Na_PIC and TDR_PIC all contributed to upregulation of excitability of 5-HT neurons. 5-HT (15 μM) enhanced the PICs, including a 26% increase in amplitude of the compound currents of Ca_PIC and TDR_PIC (P < 0.001, n = 9), 3.6 ± 5 mV hyperpolarization of Na_PIC and TDR_PIC onset (P < 0.05, n = 12), 30% increase in amplitude of TDR_PIC (P < 0.01), and 2.0 ± 3 mV hyperpolarization of TDR_PIC onset (P < 0.05, n = 18). 5-HT also facilitated repetitive firing of 5-HT neurons through modulation of composite PIC, Na_PIC and TDR_PIC, and Ca_PIC and TDR_PIC, respectively. In particular, the high voltage-activated TDR_PIC facilitated the repetitive firing in higher membrane potential, and this facilitation could be amplified by 5-HT. Morphological data analysis indicated that the dendrites of 5-HT neurons possessed dense spherical varicosities intensively crossing 5-HT neurons in medulla. We characterized the PICs in 5-HT neurons and unveiled the mechanism underlying upregulation of excitability of 5-HT neurons through serotonergic modulation of PICs. This study provided insight into channel mechanisms responsible for the serotonergic modulation of serotonergic neurons in brainstem.
Variants of Novel Immunomodulatory Fc Receptor Like 5 Gene Are Associated With Multiple Sclerosis Susceptibility in the Polish Population
Authors: Chorazy Monika , Wawrusiewicz-Kurylonek Natalia , Adamska-Patruno Edyta , Czarnowska Agata , Zajkowska Olga , Kapica-Topczewska Katarzyna , Posmyk Renata , Kretowski Adam Jacek , Kochanowicz Jan , Kułakowska Alina
Published: April 2021

Fc receptors have been shown to play a role in several autoimmune diseases. We aimed to test, for the first time, whether some of the single nucleotide variants in the FCRL5 gene were associated with multiple sclerosis (MS) susceptibility and clinical manifestations in the Polish population. The case-control study included 94 individuals with MS and 160 healthy subjects. We genotyped two single nucleotide variants of the FCRL5 gene: rs2012199 and rs6679793. The age of onset, disease duration, and clinical condition of the MS subjects were analyzed. For statistical analysis, we used the chi-squared test confirmed with Fisher's exact test. We observed the significant differences in the distribution of investigated FCRL5 genotypes between MS subjects and healthy controls. The CC and CT genotypes, as well as the C allele of rs2012199, were significantly more common in the MS subjects, as were genotypes AA and AG, and allele A of rs6679793. We noted that decreased MS susceptibility was associated with the T allele rs2012199 (OR = 0.37, p = 0.0002) and G allele rs6679793 (OR = 0.6, p = 0.02). Our results support the role of the FCRL5 locus in MS predisposition and extend the evidence of its influence on autoimmunity.
Current Trends, Controversies, and Future Directions in the Evaluation and Management of Superior Canal Dehiscence Syndrome
Authors: Eberhard Kristine Elisabeth , Chari Divya A. , Nakajima Hideko Heidi , Klokker Mads , Cayé-Thomasen Per , Lee Daniel J.
Published: April 2021

Patients with superior canal dehiscence syndrome (SCDS) can present with a range of auditory and/or vestibular signs and symptoms that are associated with a bony defect of the superior semicircular canal (SSC). Over the past two decades, advances in diagnostic techniques have raised the awareness of SCDS and treatment approaches have been refined to improve patient outcomes. However, a number of challenges remain. First, there is currently no standardized clinical testing algorithm for quantifying the effects of superior canal dehiscence (SCD). SCDS mimics a number of common otologic disorders and established metrics such as supranormal bone conduction thresholds and vestibular evoked myogenic potential (VEMP) measurements; although useful in certain cases, have diagnostic limitations. Second, while high-resolution computed tomography (CT) is the gold standard for the detection of SCD, a bony defect does not always result in signs and symptoms. Third, even when SCD repair is indicated, there is a lack of consensus about nomenclature to describe the SCD, ideal surgical approach, specific repair techniques, and type of materials used. Finally, there is no established algorithm in evaluation of SCDS patients who fail primary repair and may be candidates for revision surgery. Herein, we will discuss both contemporary and emerging diagnostic approaches for patients with SCDS and highlight challenges and controversies in the management of this unique patient cohort.
The Potential Role of Herpes Simplex Virus Type 1 and Neuroinflammation in the Pathogenesis of Alzheimer's Disease
Authors: Laval Kathlyn , Enquist Lynn W.
Published: April 2021

Alzheimer's disease (AD) is a neurodegenerative disease affecting ~50 million people worldwide. To date, there is no cure and current therapies have not been effective in delaying disease progression. Therefore, there is an urgent need for better understanding of the pathogenesis of AD and to rethink possible therapies. Herpes simplex virus type 1 (HSV1) has recently received growing attention for its potential role in sporadic AD. The virus is a ubiquitous human pathogen that infects mucosal epithelia and invades the peripheral nervous system (PNS) of its host to establish a reactivable, latent infection. Upon reactivation, HSV1 spreads back to the epithelium and initiates a new infection, causing epithelial lesions. Occasionally, the virus spreads from the PNS to the brain after reactivation. In this review, we discuss current work on the pathogenesis of AD and summarize research results that support a potential role for HSV1 in the infectious hypothesis of AD. We also highlight recent findings on the neuroinflammatory response, which has been proposed to be the main driving force of AD, starting early in the course of the disease. Relevant rodent models to study neuroinflammation in AD and novel therapeutic approaches are also discussed. Throughout this review, we focus on several aspects of HSV1 pathogenesis, including its primary role as an invader of the PNS, that should be considered in the etiology of AD. We also point out some of the contradictory data and remaining knowledge gaps that require further research to finally fully understand the cause of AD in humans.
Time in Associative Learning: A Review on Temporal Maps
Authors: Chandran Midhula , Thorwart Anna
Published: April 2021

Ability to recall the timing of events is a crucial aspect of associative learning. Yet, traditional theories of associative learning have often overlooked the role of time in learning association and shaping the behavioral outcome. They address temporal learning as an independent and parallel process. Temporal Coding Hypothesis is an attempt to bringing together the associative and non-associative aspects of learning. This account proposes temporal maps, a representation that encodes several aspects of a learned association, but attach considerable importance to the temporal aspect. A temporal map helps an agent to make inferences about missing information by applying an integration mechanism over a common element present in independently acquired temporal maps. We review the empirical evidence demonstrating the construct of temporal maps and discuss the importance of this concept in clinical and behavioral interventions.
Lipid Extract From a Vegetable (Sonchus Oleraceus) Attenuates Adipogenesis and High Fat Diet-Induced Obesity Associated With AMPK Activation
Authors: Chen Chih-Yu , Su Chien-Wen , Li Xiangyong , Liu Yinghua , Pan Qian , Cao Tinglan , Kang Jing X.
Published: April 2021

Scope:Sonchus Oleraceus, named bitter vegetable (BV), has been known to have multiple health benefits such as anti-aging and anti-inflammation. However, the role of BV in the prevention of obesity is unclear. The aim of this study was to examine the effect of BV lipid extracts (BVL) on obesity development.

Methods and Results: Following treatments of high fat diet-induced obese mice (C57BL/6J) with BVL (0.3 mg/g of BW per mouse) for a month, mice exhibited a significant reduction in weight gain, blood triglyceride, and fasting blood glucose compared to control mice. Intriguingly, phosphorylated AMPK, a key regulator of nutrient metabolism, was markedly increased in inguinal fat of BVL group. In 3T3-L1 cells, BVL-7 (100 μg/ml), an omega-3 fatty acid-rich fraction from BVL, lowered lipid accumulation, and down-regulated the gene expression of adipocyte markers. The inhibitory effect of BVL occurred at the early stage of adipocyte differentiation, leading to the delay of mitotic clonal expansion. AMPK knockdown by siRNA abolished the inhibitory effect of BVL-7 on adipogenesis, suggesting that AMPK is essential for BVL-regulated adipocyte differentiation.

Conclusion: BVL can effectively inhibit adipogenesis through, at least in part, stimulating AMPK pathway and attenuate HFD-induced obesity. Our findings suggest that BVL can be a promising dietary supplement for protection against obesity, and the effective component of BVL can be potentially developed as anti-obesity drugs.
AGAP2-AS1 May Promote the Occurrence and Development of Glioblastoma by Sponging miR-9-5p: Evidence From a ceRNA Network
Authors: Luo Xiaobin , Tu Tianqi , Zhong Yali , Xu Shangyi , Chen Xiangzhou , Chen Ligang , Yang Fubing
Published: April 2021

Glioblastoma (GBM), the primary malignant brain tumor, is typically associated with a poor prognosis and poor quality of life, mainly due to the lack of early diagnostic biomarkers and therapeutic targets. However, gene sequencing technologies and bioinformatics analysis are currently being actively utilized to explore potential targets for the diagnosis and management of malignancy. Herein, based on a variety of bioinformatics tools for the reverse prediction of target genes associated with the prognosis of GBM, a ceRNA network of AGAP2-AS1-miR-9-5p-MMP2/MMP9 was constructed, and a potential therapeutic target for GBM was identified. Enrichment analysis predicted that the ceRNA regulatory network participates in the processes of cell proliferation, differentiation, and migration.
Case Report: Cetuximab in Combination With Chemotherapy for the Treatment of Multifocal Hepatic Metastases From Colorectal Cancer Guided by Genetic Tests
Authors: Qiu Chunhui , Xie Sidong , Cheng Na , Lin Qu , Shen Guanzhu , Xiang Zhanwang , Huang Tanxiao , Zhang Xiaoni , Duan Jingxian , Wei Li , Zheng Zongheng
Published: April 2021

Hepatic metastases were reported in up to 70% of colorectal cancer patients, among which multifocal hepatic metastasis represents one of the complications that lead to poor prognosis. The majority of the patients carrying multifocal hepatic metastases required pharmaceutical treatments to reduce the tumor size prior to surgical resection. However, the clinical responses to pharmaceutical agents were difficult to predict due to the heterogeneous nature of the multifocal tumors. Here, we report a case with multifocal hepatic metastases from colorectal cancer that was resistant to the primary chemotherapy and Bevacizumab plus chemotherapy, but responded to the combined therapy of Cetuximab and FOLFOX. Genetic tests had revealed that the tumor was highly metastatic due to the mutations of the WNT signaling pathway, and the metastatic tumors might be sensitive to Cetuximab. Consistent with the molecular characterizations, the metastatic tumors continue to emerge after chemotherapy, and rapidly relapsed in great numbers after liver resection. However, the combined therapy of Cetuximab and FOLFOX guided by the genetic tests significantly reduced the size and number of metastatic tumors. To conclude, deciphering the mutation profiles of multifocal metastatic tumors may guide the determination of treatment tactics, which may benefit the patients with non-resectable advanced carcinoma.
Identification of Novel Prognostic Risk Signatures of Soft Tissue Sarcoma Based on Ferroptosis-Related Genes
Authors: Huang Wenjing , Duan Yuhe , Yang Xiuwei , Shang Cong , Chen Xin , Zhang Huanyu , Li Fujiang
Published: April 2021

Background

The role of ferroptosis in tumorigenesis has been confirmed in previous studies. However, the comprehensive analysis of ferroptosis-related gene (FRG) to study the role of FRG in soft tissue sarcoma (STS) is lacking.

Methods

RNA sequencing profile of TCGA-SARC cohort and GTEx were used to select differentially expressed FRGs (DEFRGs). Univariate, LASSO, and multivariate Cox analyses were selected to determine overall survival (OS)- and disease-free survival (PFS)-related FRGs. Two prognostic signatures were established and validated in two independent sets from Gene Expression Omnibus (GEO). Finally, the expression of key FRGs were validated with RT-qPCR.

Results

In total, 198 FRGs (90.4%) were abnormally expressed in STS. Twelve DEFRGs were incorporated in the final signatures and showed favorable discrimination in both training and validation cohorts. Patients in the different risk groups not only showed different prognosis, but also showed different infiltration of immune cells. Two nomograms combining signature and clinical variables were established and the C-indexes were 0.852 and 0.752 for the OS and DFS nomograms, respectively. Finally, the expression of NOX5, HELLS, and RPL8 were validated with RT-qPCR.

Conclusion

This comprehensive analysis of the FRG landscape in STS revealed novel FRGs related to carcinogenesis and prognosis. These findings have implications for prognosis and therapeutic responses, which revealed potential prognostic biomarkers and promote precision medicine.
Comprehensive Study of Germline Mutations and Double-Hit Events in Esophageal Squamous Cell Cancer
Authors: Zeng Bing , Huang Peide , Du Peina , Sun Xiaohui , Huang Xuanlin , Fang Xiaodong , Li Lin
Published: April 2021

Esophageal squamous cell cancer (ESCC) is the eighth most common cancer around the world. Several reports have focused on somatic mutations and common germline mutations in ESCC. However, the contributions of pathogenic germline alterations in cancer susceptibility genes (CSGs), highly frequently mutated CSGs, and pathogenically mutated CSG-related pathways in ESCC remain unclear. We obtained data on 571 ESCC cases from public databases and East Asian from the 1000 Genomes Project database and the China Metabolic Analytics Project database to characterize pathogenic mutations. We detected 157 mutations in 75 CSGs, accounting for 25.0% (143/571) of ESCC cases. Six genes had more than five mutations: TP53 (n = 15 mutations), GJB2 (n = 8), BRCA2 (n = 6), RECQL4 (n = 6), MUTYH (n = 6), and PMS2 (n = 5). Our results identified significant differences in pathogenic germline mutations of TP53, BRCA2, and RECQL4 between the ESCC and control cohorts. Moreover, we identified 84 double-hit events (16 germline/somatic double-hit events and 68 somatic/somatic double-hit events) occurring in 18 tumor suppressor genes from 83 patients. Patients who had ESCC with germline/somatic double-hit events were diagnosed at younger ages than patients with the somatic/somatic double-hit events, though the correlation was not significant. Fanconi anemia was the most enriched pathway of pathogenically mutated CSGs, and it appeared to be a primary pathway for ESCC predisposition. The results of this study identified the underlying roles that pathogenic germline mutations in CSGs play in ESCC pathogenesis, increased our awareness about the genetic basis of ESCC, and provided suggestions for using highly mutated CSGs and double-hit features in the early discovery, prevention, and genetic counseling of ESCC.
Apigenin Inhibits the Growth of Hepatocellular Carcinoma Cells by Affecting the Expression of microRNA Transcriptome
Authors: Wang Shou-Mei , Yang Pei-Wei , Feng Xiao-Jun , Zhu Yi-Wei , Qiu Feng-Jun , Hu Xu-Dong , Zhang Shu-Hui
Published: April 2021

Background

Apigenin, as a natural flavonoid, has low intrinsic toxicity and has potential pharmacological effects against hepatocellular carcinoma (HCC). However, the molecular mechanisms involving microRNAs (miRNAs) and their target genes regulated by apigenin in the treatment of HCC have not been addressed.

Objective

In this study, the molecular mechanisms of apigenin involved in the prevention and treatment of HCC were explored in vivo and in vitro using miRNA transcriptomic sequencing to determine the basis for the clinical applications of apigenin in the treatment of HCC.

Methods

The effects of apigenin on the proliferation, cell cycle progression, apoptosis, and invasion of human hepatoma cell line Huh7 and Hep3B were studied in vitro, and the effects on the tumorigenicity of Huh7 cells were assessed in vivo. Then, a differential expression analysis of miRNAs regulated by apigenin in Huh7 cells was performed using next-generation RNA sequencing and further validated by qRT-PCR. The potential genes targeted by the differentially expressed miRNAs were identified using a curated miRTarBase miRNA database and their molecular functions were predicted using Gene Ontology and KEGG signaling pathway analysis.

Results

Compared with the control treatment group, apigenin significantly inhibited Huh7 cell proliferation, cell cycle, colony formation, and cell invasion in a concentration-dependent manner. Moreover, apigenin reduced tumor growth, promoted tumor cell necrosis, reduced the expression of Ki67, and increased the expression of Bax and Bcl-2 in the xenograft tumors of Huh7 cells. Bioinformatics analysis of the miRNA transcriptome showed that hsa-miR-24, hsa-miR-6769b-3p, hsa-miR-6836-3p, hsa-miR-199a-3p, hsa-miR-663a, hsa-miR-4739, hsa-miR-6892-3p, hsa-miR-7107-5p, hsa-miR-1273g-3p, hsa-miR-1343, and hsa-miR-6089 were the most significantly up-regulated miRNAs, and their key gene targets were MAPK1, PIK3CD, HRAS, CCND1, CDKN1A, E2F2, etc. The core regulatory pathways of the up-regulated miRNAs were associated with the hepatocellular carcinoma pathway. The down-regulated miRNAs were hsa-miR-181a-5p and hsa-miR-148a-3p, and the key target genes were MAPK1, HRAS, STAT3, FOS, BCL2, SMAD2, PPP3CA, IFNG, MET, and VAV2, with the core regulatory pathways identified as proteoglycans in cancer pathway.

Conclusion

Apigenin can inhibit the growth of HCC cells, which may be mediated by up-regulation or down-regulation of miRNA molecules and their related target genes.