bluuebunny/crossref_metadata_2025 huggingface dataset:
Ulcerative Colitis-Derived Colonoid Culture: A Multi-Mineral-Approach to Improve Barrier Protein Expression
Authors: Aslam Muhammad N. , McClintock Shannon D. , Attili Durga , Pandya Shailja , Rehman Humza , Nadeem Daniyal M. , Jawad-Makki Mohamed Ali H. , Rizvi Areeba H. , Berner Maliha M. , Dame Michael K. , Turgeon Danielle Kim , Varani James
Published: November 2020

Background

Recent studies demonstrated that Aquamin®, a calcium-, magnesium-rich, multi-mineral natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved in tissue from subjects with ulcerative colitis (UC).

Methods

Colonoid cultures were established with colon biopsies from 9 individuals with UC. The colonoids were then incubated for a 2-week period under control conditions (in culture medium with a final calcium concentration of 0.25 mM) or in the same medium supplemented with Aquamin®to provide 1.5 – 4.5 mM calcium. Effects on differentiation and barrier protein expression were determined using several approaches: phase-contrast and scanning electron microscopy, quantitative histology and immunohistology, mass spectrometry-based proteome assessment and transmission electron microscopy.

Results

Although there were no gross changes in colonoid appearance, there was an increase in lumen diameter and wall thickness on histology and greater expression of cytokeratin 20 (CK20) along with reduced expression of Ki67 by quantitative immunohistology observed with intervention. In parallel, upregulation of several differentiation-related proteins was seen in a proteomic screen with the intervention. Aquamin®-treated colonoids demonstrated a modest up-regulation of tight junctional proteins but stronger induction of adherens junction and desmosomal proteins. Increased desmosomes were seen at the ultrastructural level. Proteomic analysis demonstrated increased expression of several basement membrane proteins and hemidesmosomal components. Proteins expressed at the apical surface (mucins and trefoils) were also increased as were several additional proteins with anti-microbial activity or that modulate inflammation. Finally, several transporter proteins that affect electrolyte balance (and, thereby affect water resorption) were increased. At the same time, growth and cell cycle regulatory proteins (Ki67, nucleophosmin, and stathmin) were significantly down-regulated. Laminin interactions, matrix formation and extracellular matrix organization were the top three up-regulated pathways with the intervention.

Conclusion

A majority of individuals including patients with UC do not reach the recommended daily intake for calcium and other minerals. To the extent that such deficiencies might contribute to the weakening of the colonic barrier, the findings employing UC tissue-derived colonoids here suggest that adequate mineral intake might improve the colonic barrier.
Erythroid Differentiation and Heme Biosynthesis Are Dependent on a Shift in the Balance of Mitochondrial Fusion and Fission Dynamics
Authors: Gonzalez-Ibanez Alvaro M. , Ruiz Lina M. , Jensen Erik , Echeverria Cesar A. , Romero Valentina , Stiles Linsey , Shirihai Orian S. , Elorza Alvaro A.
Published: November 2020

Erythropoiesis is the most robust cellular differentiation and proliferation system, with a production of ∼2 × 1011 cells per day. In this fine-tuned process, the hematopoietic stem cells (HSCs) generate erythroid progenitors, which proliferate and mature into erythrocytes. During erythropoiesis, mitochondria are reprogrammed to drive the differentiation process before finally being eliminated by mitophagy. In erythropoiesis, mitochondrial dynamics (MtDy) are expected to be a key regulatory point that has not been described previously. We described that a specific MtDy pattern occurs in human erythropoiesis from EPO-induced human CD34+ cells, characterized predominantly by mitochondrial fusion at early stages followed by fission at late stages. The fusion protein MFN1 and the fission protein FIS1 are shown to play a key role in the progression of erythropoiesis. Fragmentation of the mitochondrial web by the overexpression of FIS1 (gain of fission) resulted in both the inhibition of hemoglobin biosynthesis and the arrest of erythroid differentiation, keeping cells in immature differentiation stages. These cells showed specific mitochondrial features as compared with control cells, such as an increase in round and large mitochondrial morphology, low mitochondrial membrane potential, a drop in the expression of the respiratory complexes II and IV and increased ROS. Interestingly, treatment with the mitochondrial permeability transition pore (mPTP) inhibitor, cyclosporin A, rescued mitochondrial morphology, hemoglobin biosynthesis and erythropoiesis. Studies presented in this work reveal MtDy as a hot spot in the control of erythroid differentiation, which might signal downstream for metabolic reprogramming through regulation of the mPTP.
A Rapid and Simple Method for Lithium Purification and Isotopic Analysis of Geological Reference Materials by MC-ICP-MS
Authors: Zhu Guanhong , Ma Jinlong , Wei Gangjian , Zhang Le
Published: November 2020

A simple method has been developed to purify lithium (Li) from matrix elements in geological reference materials, using a single-column packed with AGMP-50 cation exchange resin, followed by high-precision Li isotope measurements by multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS). A series of tests, such as different types of resin, loading amount of Li, loading volumes, and various eluents, were conducted to ascertain the optimal conditions for Li purification and the effects of intensity, acidity, and presence of potential matrix elements on Li isotope measurements were also evaluated. In our experiment, Al and high-field-strength elements (HFSEs), such as Ti, Zr, and Hf, were eluted by 0.2 M HCl + 0.3 M HF, and 0.73 M HCl was used to separate Li from other matrix elements, such as Na. This method is suitable for processing large amount of Li (60–270 ng) and enabling a Li recovery of close to 100%, with effective removal of matrix elements such as Na and Ca. Besides, our method achieves low matrix interferences (e.g., Na/Li << 1 and Ca/Li << 1 for rock and seawater via a single-column procedure; Ca/Li < 2 for carbonate via a two-column procedure) and also uses small volume of eluents and is rapid (~5 h), enabling a total separation to be completed in ~0.5 d. Using this method, we report Li isotopic compositions of various geological reference materials, including igneous rocks, seawater, and carbonate. The Li isotopic compositions are consistent with the data published previously for the analyzed reference materials. As such, the reported method is ideally suited for Li separation from multiple types of geological samples prior to isotopic analysis.
Structural Insights on Tiny Peptide Nucleic Acid (PNA) Analogues of miRNA-34a: An in silico and Experimental Integrated Approach
Authors: Moccia Maria , Mercurio Flavia Anna , Langella Emma , Piacenti Valerio , Leone Marilisa , Adamo Mauro F. A. , Saviano Michele
Published: November 2020

In the present work, structural features of the interaction between peptide nucleic acid (PNA)-based analogs of the tumor-suppressor microRNA-34a with both its binding sites on MYCN mRNA were investigated. In particular, the region from base 1 to 8 (“seed” region) of miR-34a was reproduced in the form of an 8-mer PNA fragment (tiny PNA), and binding to target 3'UTR MYCN mRNA, was studied by a seldom reported and detailed NMR characterization, providing evidence for the formation of anti-parallel duplexes with a well-organized structural core. The formation of PNA-3'UTR duplexes was also confirmed by Circular Dichroism, and their melting curves were measured by UV spectroscopy. Nevertheless, this study offered a valuable comparison between molecular dynamics predictions and experimental evidence, which showed great correlation. Preliminary uptake assays were carried out in Neuroblastoma Kelly cells, using short peptide conjugates as carriers and FITC fluorescent tag for subcellular localization. Moderate internalization was observed without the use of transfecting agents. The reported results corroborate the interest toward the design and development of chimeric PNA/RNA sequences as effective RNA-targeting agents.
Use of Natural Products in Leishmaniasis Chemotherapy: An Overview
Authors: Gervazoni Luiza F. O. , Barcellos Gabrielle B. , Ferreira-Paes Taiana , Almeida-Amaral Elmo E.
Published: November 2020

Leishmaniasis is an infectious parasitic disease that is caused by protozoa of the genus Leishmania, a member of the Trypanosomatidae family. Leishmaniasis is classified by the World Health Organization as a neglected tropical disease that is responsible for millions of deaths worldwide. Although there are many possible treatments for leishmaniasis, these treatments remain mostly ineffective, expensive, and long treatment, as well as causing side effects and leading to the development of resistance. For novel and effective treatments to combat leishmaniasis, many research groups have sought to utilize natural products. In addition to exhibiting potential as therapeutic compounds, natural products may also contribute to the development of new drugs based on their chemical structures. This review presents the most promising natural products, including crude extracts and isolated compounds, employed against Leishmania spp.
Nanoparticle Formulations of Poly (ADP-ribose) Polymerase Inhibitors for Cancer Therapy
Authors: Singh Bijay , Yang Shicheng , Krishna Apurva , Sridhar Srinivas
Published: November 2020

A number of poly(ADP-ribose) polymerase (PARP) inhibitors have been recently approved for clinical use in BRCA mutated and other cancers. However, off-target toxicity of PARP inhibitors and the emergence of drug resistance following prolonged administration of these inhibitors indicate the need for improved methods of drug delivery to the tumors. Nanomedicines based upon nanoparticle formulations of conventional small molecule drugs and inhibitors offer many advantages, such as increased solubility and bioavailability of drugs, reduced toxicity and drug resistance, and improved tissue selectivity and therapeutic efficacy. This review highlights the current trends in formulations of PARP inhibitors developed by nanotechnology approaches and provides an insight into the applications and limitations of these PARP inhibitor nanomedicines for cancer therapies.
Extreme Natural Events Mitigation: An Analysis of the National Disaster Funds in Latin America
Authors: Szlafsztein Claudio Fabian
Published: November 2020

This paper reviews, compares, and analyzes the legal and institutional framework of Latin American government insurance systems for disaster risk. Data and information are obtained through an intense examination of disasters database, the scientific literature and legal framework of administrative and operational procedures, and mainly sources of financial funds related to disaster risk management. The results demonstrate that all countries, with the exception of Ecuador and Chile, legally establish some form of fund by their own legislation and regulation, as a total or partial form of financing the management of natural disasters, particularly those classified as post-disaster recuperation and reconstruction practices. North and Central American countries have more complex and well-structured funds, presumably based on their history of natural disasters and high social vulnerability. The funds are composed of initial values defined by law, annual contributions of general budgets, donations, and the financial gains of resources deposited into bank accounts. The paper concludes that the unavailability of required resources in an emergency situation has led Latin American countries to choose disaster funds as a primary disaster risk management financing strategy. However, the uncertainty of natural disaster occurrence is one of the main obstacles to the use of public financial resources in prevention and preparedness strategies and actions, increasing even more with the inclusion of climate change uncertainties.
Machine Learning Methods to Predict Acute Respiratory Failure and Acute Respiratory Distress Syndrome
Authors: Wong An-Kwok Ian , Cheung Patricia C. , Kamaleswaran Rishikesan , Martin Greg S. , Holder Andre L.
Published: November 2020

Acute respiratory failure (ARF) is a common problem in medicine that utilizes significant healthcare resources and is associated with high morbidity and mortality. Classification of acute respiratory failure is complicated, and it is often determined by the level of mechanical support that is required, or the discrepancy between oxygen supply and uptake. These phenotypes make acute respiratory failure a continuum of syndromes, rather than one homogenous disease process. Early recognition of the risk factors for new or worsening acute respiratory failure may prevent that process from occurring. Predictive analytical methods using machine learning leverage clinical data to provide an early warning for impending acute respiratory failure or its sequelae. The aims of this review are to summarize the current literature on ARF prediction, to describe accepted procedures and common machine learning tools for predictive tasks through the lens of ARF prediction, and to demonstrate the challenges and potential solutions for ARF prediction that can improve patient outcomes.
Advancing Field-Based GNSS Surveying for Validation of Remotely Sensed Water Surface Elevation Products
Authors: Pitcher Lincoln H. , Smith Laurence C. , Cooley Sarah W. , Zaino Annie , Carlson Robert , Pettit Joseph , Gleason Colin J. , Minear J. Toby , Fayne Jessica V. , Willis Michael J. , Hansen Jasmine S. , Easterday Kelly J. , Harlan Merritt E. , Langhorst Theodore , Topp Simon N. , Dolan Wayana , Kyzivat Ethan D. , Pietroniro Al , Marsh Philip , Yang Daqing , Carter Tom , Onclin Cuyler , Hosseini Nasim , Wilcox Evan , Moreira Daniel , Berge-Nguyen Muriel , Cretaux Jean-Francois , Pavelsky Tamlin M.
Published: November 2020

To advance monitoring of surface water resources, new remote sensing technologies including the forthcoming Surface Water and Ocean Topography (SWOT) satellite (expected launch 2022) and its experimental airborne prototype AirSWOT are being developed to repeatedly map water surface elevation (WSE) and slope (WSS) of the world’s rivers, lakes, and reservoirs. However, the vertical accuracies of these novel technologies are largely unverified; thus, standard and repeatable field procedures to validate remotely sensed WSE and WSS are needed. To that end, we designed, engineered, and operationalized a Water Surface Profiler (WaSP) system that efficiently and accurately surveys WSE and WSS in a variety of surface water environments using Global Navigation Satellite Systems (GNSS) time-averaged measurements with Precise Point Positioning corrections. Here, we present WaSP construction, deployment, and a data processing workflow. We demonstrate WaSP data collections from repeat field deployments in the North Saskatchewan River and three prairie pothole lakes near Saskatoon, Saskatchewan, Canada. We find that WaSP reproducibly measures WSE and WSS with vertical accuracies similar to standard field survey methods [WSE root mean squared difference (RMSD) ∼8 cm, WSS RMSD ∼1.3 cm/km] and that repeat WaSP deployments accurately quantify water level changes (RMSD ∼3 cm). Collectively, these results suggest that WaSP is an easily deployed, self-contained system with sufficient accuracy for validating the decimeter-level expected accuracies of SWOT and AirSWOT. We conclude by discussing the utility of WaSP for validating airborne and spaceborne WSE mappings, present 63 WaSP in situ lake WSE measurements collected in support of NASA’s Arctic-Boreal and Vulnerability Experiment, highlight routine deployment in support of the Lake Observation by Citizen Scientists and Satellites project, and explore WaSP utility for validating a novel GNSS interferometric reflectometry LArge Wave Warning System.
Therapeutic Perspectives on the Modulation of G-Protein Coupled Estrogen Receptor, GPER, Function
Authors: Rouhimoghadam Milad , Lu Anh S. , Salem Aliasger K. , Filardo Edward J.
Published: November 2020

Estrogens exert their physiological and pathophysiological effects via cellular receptors, named ERα, ERβ, and G-protein coupled estrogen receptor (GPER). Estrogen-regulated physiology is tightly controlled by factors that regulate estrogen bioavailability and receptor sensitivity, while disruption of these control mechanisms can result in loss of reproductive function, cancer, cardiovascular and neurodegenerative disease, obesity, insulin resistance, endometriosis, and systemic lupus erythematosus. Restoration of estrogen physiology by modulating estrogen bioavailability or receptor activity is an effective approach for treating these pathological conditions. Therapeutic interventions that block estrogen action are employed effectively for the treatment of breast and prostate cancer as well as for precocious puberty and anovulatory infertility. Theoretically, treatments that block estrogen biosynthesis should prevent estrogen action at ERs and GPER, although drug resistance and ligand-independent receptor activation may still occur. In addition, blockade of estrogen biosynthesis does not prevent activation of estrogen receptors by naturally occurring or man-made exogenous estrogens. A more complicated scenario is provided by anti-estrogen drugs that antagonize ERs since these drugs function as GPER agonists. Based upon its association with metabolic dysregulation and advanced cancer, GPER represents a therapeutic target with promise for the treatment of several critical health concerns facing Western society. Selective ligands that specifically target GPER have been developed and may soon serve as pharmacological agents for treating human disease. Here, we review current forms of estrogen therapy and the implications that GPER holds for these therapies. We also discuss existing GPER targeted drugs, additional approaches towards developing GPER-targeted therapies and how these therapies may complement existing modalities of estrogen-targeted therapy.
Risk Assessment Regarding Perceived Toxicity and Acceptance of Carbon Dioxide-Based Fuel by Laypeople for Its Use in Road Traffic and Aviation
Authors: Engelmann Linda , Arning Katrin , Linzenich Anika , Ziefle Martina
Published: November 2020

One approach to mitigate the emissions of carbon dioxide (CO2) is the development of CO2-based products, such as fuels for road traffic and aviation. Since the acceptance of sustainable product innovations such as CO2-based fuels depends on an individual’s acceptance decision based on perceived risks and benefits, this study focuses on subjective risk perceptions of fuel toxicity. An online survey was conducted to assess risk evaluations of CO2-based fuels regarding various risk targets, exposure characteristics, negative outcomes for health and environment, and frequency of health impairments. CO2-based fuels were significantly more positively perceived than conventional fuels and were found to be perceived to pose less risks regarding types of exposure and properties leading to toxic effects. For both aviation and road traffic the acceptance of CO2-based fuels increased with decreasing fear of health and environmental consequences and the less frequently health effects were assessed. The findings allow to derive implications for risk assessment and communication strategies in the development and roll-out of CO2-based fuels.
Numerical Simulation of a Horizontal Well With Multi-Stage Oval Hydraulic Fractures in Tight Oil Reservoir Based on an Embedded Discrete Fracture Model
Authors: Yang Zhi-dong , Wang Yong , Zhang Xu-yang , Qin Ming , Su Shao-wen , Yao Zhen-hua , Liu Lingfu
Published: November 2020

Tight oil is a kind of unconventional oil and gas resource with great development potential. Due to the unconventional characteristics of low porosity and low permeability in tight oil reservoirs, single wells generally have no natural productivity, and industrial development is usually conducted in combination with horizontal wells and hydraulic fracturing techniques. To capture the flow behavior affected by fractures with complex geometry and interaction, we adopted embedded discrete fracture models (EDFMs) to simulate the development of fractured reservoirs. Compared with the traditional discrete fracture models (DFMs), the embedded discrete fracture models (EDFMs) can not only accurately represent the fracture geometry but also do not generate a large number of refine grids around fractures and intersections of fractures, which shows the high computational efficiency. To be more consistent with the real characteristic of the reservoir and reflect the advantage of EDFMs on modeling complex fractures, in this work, the hydraulic fractures are set as oval shape, and we adopted 3-dimensional oil–gas two-phase model considering capillary forces and gravity effects. We developed an EDFM simulator, which is verified by using the fine grid method (FGM). Finally, we simulated and studied the development of tight oil without and with random natural fractures (NFs). In our simulation, the pressure varies widely from the beginning to the end of the development. In real situation, tight oil reservoirs have high initial pressure and adopt step-down bottom hole pressure development strategy where the bottom hole pressure of the last stage is below the bubble point pressure and the free gas appears in the reservoir. Modeling studies indicate that the geometry of fracture has a great influence on the pressure and saturation profiles in the area near the fractures, and dissolved gas flooding contributes to the development of tight oil, and NFs can significantly improve production, while the effect of the stress sensitivity coefficient of NFs on production is more significant in the later stage of production with lower reservoir pressure.
Pronounced Fidelity and Selection for Average Conditions of Calving Area Suggestive of Spatial Memory in a Highly Migratory Ungulate
Authors: Cameron Matthew D. , Joly Kyle , Breed Greg A. , Mulder Christa P. H. , Kielland Knut
Published: November 2020

A distinguishing characteristic of many migratory animals is their annual return to distinct calving (birthing) areas in the spring, yet the navigational mechanisms employed during migration that result in this pattern are poorly understood. Effective conservation of these species requires reliable delineation of such areas, quantifying the factors that influence their selection, and understanding the underlying mechanisms resulting in use of calving areas. We used barren-ground caribou (Rangifer tarandus granti) as a study species and identified calving sites of the Western Arctic Herd in Alaska using GPS collar data from 2010–2017. We assessed variability in calving areas by comparing spatial delineations across all combinations of years. To understand calving area selection at a landscape scale, we performed a resource selection analysis comparing calving sites to available locations across the herd’s range and incorporated time-varying, remotely sensed metrics of vegetation quality and quantity. We found that whereas calving areas varied from year to year, this annual variation was centered on an area of recurring attraction consistent with previous studies covering the last six decades. Calving sites were characterized by high-quality forage at the average time of calving, but not peak calving that year, and by a narrow range of distinct physiographic factors. Each year, calving sites were located on areas of above-average conditions based on our predictive model. Our findings indicate that the pattern of spring migration for pregnant females was to migrate to areas that consistently provide high-quality forage when averaged across years, and then upon arriving at this calving ground, refine selection using their perception of annually varying conditions that are driven by environmental stochasticity. We suggest that the well-documented and widespread pattern of fidelity to calving grounds by caribou is supportive of a navigational mechanism based on spatial memory at a broad scale to optimize foraging and energy acquisition at a critical life-history stage. The extent to which migrants depend on memory to reach their spring destinations has implications for the adaptability of populations to changing climate and human impacts.
Transcription Factors of the Alx Family: Evolutionarily Conserved Regulators of Deuterostome Skeletogenesis
Authors: Khor Jian Ming , Ettensohn Charles A.
Published: November 2020

Members of thealxgene family encode transcription factors that contain a highly conserved Paired-class, DNA-binding homeodomain, and a C-terminal OAR/Aristaless domain. Phylogenetic and comparative genomic studies have revealed complex patterns ofalxgene duplications during deuterostome evolution. Remarkably,alxgenes have been implicated in skeletogenesis in both echinoderms and vertebrates. In this review, we provide an overview of current knowledge concerningalxgenes in deuterostomes. We highlight their evolutionarily conserved role in skeletogenesis and draw parallels and distinctions between the skeletogenic gene regulatory circuitries of diverse groups within the superphylum.
Serum MiR-4687-3p Has Potential for Diagnosis and Carcinogenesis in Non-small Cell Lung Cancer
Authors: Liu Man , Si Qiufang , Ouyang Songyun , Zhou Zhigang , Wang Meng , Zhao Chunling , Yang Ting , Wang Yulin , Zhang Xue , Xie Wenbo , Dai Liping , Li Jitian
Published: November 2020

The lack of a useful biomarker partly contributes to the increased mortality of non-small cell lung cancer (NSCLC). MiRNAs have become increasingly appreciated in diagnosis of NSCLC. In the present study, we used microarray to screen 2,549 miRNAs in serum samples from the training cohort (NSCLC, n = 10; the healthy, n = 10) to discover differentially expressed miRNAs (DEMs). Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assay was applied to validate the expression level of selected overexpressed DEMs of NSCLC in a validation cohort (NSCLC, n = 30; the healthy, n = 30). Area under the receiver operating characteristic curve (AUC) was performed to evaluate diagnostic capability of the DEMs. The expression of the miRNAs in tissues was analyzed based on the TCGA database. Subsequently, the target genes of the miR-4687-3p were predicted by TargetScan. Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were tested by R software (ClusterProfiler package). NSCLC cells were transfected with inhibitor or mimic to down-regulate or up-regulate the miR-4687-3p level. The function of miR-4687-3p on proliferation, invasion, and migration of lung cancer cells were investigated through CCK-8 and Transwell assays, respectively. In the results, we identified serum miR-4687-3p that provided a high diagnostic accuracy of NSCLC (AUC = 0.679, 95%CI: 0.543–0.815) in the validation cohort. According to the TCGA database, we found that the miR-4687-3p level was significantly higher in NSCLC tissues than in normal lung tissues (p < 0.05). GO and KEGG pathway enrichment analysis showed that postsynaptic specialization and TGF-β signaling pathway were significantly enriched. Down-regulation of miR-4687-3p could suppress the proliferation, invasion, and migration of the NSCLC cells, compared with inhibitor negative control (NC). Meanwhile, overexpression of miR-4687-3p could promote the proliferation, invasion, and migration of the NSCLC cells compared with mimic NC. As a conclusion, our study first discovered that serum miR-4687-3p might have clinical potential as a non-invasive diagnostic biomarker for NSCLC and play an important role in the development of NSCLC.
IL-Y Aggravates Murine Chronic Graft-Versus-Host Disease by Enhancing T and B Cell Responses
Authors: Wan Li , Jin Ziqi , Hu Bo , Lv Kangkang , Lei Lei , Liu Yonghao , Song Yuan , Zhu Ying , Gong Huanle , Xu Mimi , Du Yuanyuan , Xu Yang , Liu Haiyan , Wu Depei , Liu Yuejun
Published: November 2020

IL-Y, a synthetic member of IL-12 cytokine family, was found to exert potent immunosuppressive effects by inhibiting the differentiation and activation of Th1 and Th17 cells. However, the role of IL-Y in the development of chronic graft-versus-host disease (cGVHD) remains unknown. Here, using murine models of scleroderma-like and lupus-like cGVHD, we examined the function of IL-Y in the pathogenesis of cGVHD by hydrodynamically injecting minicircle-IL-Y expressing plasmids (MC IL-Y). In contrast with the reported immune suppressive function of IL-Y, administration of MC IL-Y enhanced cGVHD severity reflected by deteriorated multi-organ pathologic damages. In lupus-like cGVHD model, urine protein and the serum anti-dsDNA antibody (IgG) were significantly upregulated by IL-Y treatment. Further study demonstrated that IL-Y impacts both donor T and B cell response. In T cells, IL-Y inhibited the generation of CD4+Foxp3+ regulator T (Treg) cells during the development of cGVHD. IL-Y may also increase the infiltration of pathogenic TNF-α producing CD4+ and CD8+ T cells through IL-27Rα in recipient spleens, as this effect was diminished in IL-27Rα deficient T cells. Moreover, IL-Y enhanced the differentiation of ICOS+ T follicular helper (Tfh) cells. In B cells, the percentage of germinal center (GC) B cells in recipient spleens was significantly upregulated by MC IL-Y plasmid administration. The levels of co-stimulatory molecules, MHC-II and CD86, on B cells were also enhanced by IL-Y expression. Taken together, our data indicated that IL-Y promoted the process of cGVHD by activating pathogenic T and B cells.
Impact of Helminth Infections on Female Reproductive Health and Associated Diseases
Authors: Chetty Alisha , Omondi Millicent A. , Butters Claire , Smith Katherine Ann , Katawa Gnatoulma , Ritter Manuel , Layland Laura , Horsnell William
Published: November 2020

A growing body of knowledge exists on the influence of helminth infections on allergies and unrelated infections in the lung andgastrointestinal (GI) mucosa. However, the bystander effects of helminth infections on the female genital mucosa and reproductive health is understudied but important considering the high prevalence of helminth exposure and sexually transmitted infections inlow- andmiddle-incomecountries (LMICs). In this review, we explore current knowledge about the direct and systemic effects of helminth infections on unrelated diseases. We summarize host disease-controlling immunity of important sexually transmitted infections and introduce the limited knowledge of how helminths infections directly cause pathology to female reproductive tract (FRT), alter susceptibility to sexually transmitted infections and reproduction. We also review work by others on type 2 immunity in the FRT and hypothesize how these insights may guide future work to help understand how helminths alter FRT health.
A Conserved TCRβ Signature Dominates a Highly Polyclonal T-Cell Expansion During the Acute Phase of a Murine Malaria Infection
Authors: Smith Natasha L. , Nahrendorf Wiebke , Sutherland Catherine , Mooney Jason P. , Thompson Joanne , Spence Philip J. , Cowan Graeme J. M.
Published: November 2020

CD4+ αβ T-cells are key mediators of the immune response to a first Plasmodium infection, undergoing extensive activation and splenic expansion during the acute phase of an infection. However, the clonality and clonal composition of this expansion has not previously been described. Using a comparative infection model, we sequenced the splenic CD4+ T-cell receptor repertoires generated over the time-course of a Plasmodium chabaudi infection. We show through repeat replicate experiments, single-cell RNA-seq, and analyses of independent RNA-seq data, that following a first infection - within a highly polyclonal expansion - T-effector repertoires are consistently dominated by TRBV3 gene usage. Clustering by sequence similarity, we find the same dominant clonal signature is expanded across replicates in the acute phase of an infection, revealing a conserved pathogen-specific T-cell response that is consistently a hallmark of a first infection, but not expanded upon re-challenge. Determining the host or parasite factors driving this conserved response may uncover novel immune targets for malaria therapeutic purposes.
Potential Role of Phosphoglycerol Dihydroceramide Produced by Periodontal Pathogen Porphyromonas gingivalis in the Pathogenesis of Alzheimer’s Disease
Authors: Yamada Chiaki , Akkaoui Juliet , Ho Anny , Duarte Carolina , Deth Richard , Kawai Toshihisa , Nichols Frank , Lakshmana Madepalli K. , Movila Alexandru
Published: November 2020

Background

Among different types of sphingolipids produced by human cells, the possible engagement of ceramide species in the pathogenesis of Alzheimer’s disease (AD) has attracted recent attention. While ceramides are primarily generated byde novosynthesis in mammalian cells, only a limited number of bacterial species, produce ceramides, including phosphoglycerol dihydroceramide (PGDHC) that is produced by the key periodontal pathogenPorphyromonas gingivalis. Emerging evidence indicates that virulence factors produced byP. gingivalis, such as lipopolysaccharide and gingipain, may be engaged in the initiation and/or progression of AD. However, the potential role of PGDHC in the pathogenesis of AD remains unknown. Therefore, the aim of this study was to evaluate the influence of PGDHC on hallmark findings in AD.

Material and Methods

CHO-7WD10 and SH-SY-5Y cells were exposed to PGDHC and lipopolysaccharide (LPS) isolated fromP. gingivalis. Soluble Aβ42 peptide, amyloid precursor protein (APP), phosphorylated tau and senescence-associated secretory phenotype (SASP) factors were quantified using ELISA and Western blot assays.

Results

Our results indicate thatP. gingivalis (Pg)-derived PGDHC, but notPg-LPS, upregulated secretion of soluble Aβ42 peptide and expression of APP in CHO-7WD10 cells. Furthermore, hyperphosphorylation of tau protein was observed in SH-SY-5Y cells in response to PGDHC lipid. In contrast,Pg-LPS had little, or no significant effect on the tau phosphorylation induced in SH-SY-5Y cells. However, both PGDHC andPg-LPS contributed to the senescence of SH-SY5Y cells as indicated by the production of senescence-associated secretory phenotype (SASP) markers, including beta-galactosidase, cathepsin B (CtsB), and pro-inflammatory cytokines TNF-α, and IL-6. Additionally, PGDHC diminished expression of the senescence-protection marker sirtuin-1 in SH-SY-5Y cells.

Conclusions

Altogether, our results indicate thatP. gingivalis-derived PGDHC ceramide promotes amyloidogenesis and hyperphosphorylation, as well as the production of SASP factors. Thus, PGDHC may represent a novel class of bacterial-derived virulence factors for AD associated with periodontitis.
RNA Sequencing Keloid Transcriptome Associates Keloids With Th2, Th1, Th17/Th22, and JAK3-Skewing
Authors: Wu Jianni , Del Duca Ester , Espino Michael , Gontzes Alyssa , Cueto Inna , Zhang Ning , Estrada Yeriel D. , Pavel Ana B. , Krueger James G. , Guttman-Yassky Emma
Published: November 2020

Keloids are disfiguring, fibroproliferative growths and their pathogenesis remains unclear, inhibiting therapeutic development. Available treatment options have limited efficacy and harbor safety concerns. Thus, there is a great need to clarify keloid pathomechanisms that may lead to novel treatments. In this study, we aimed to elucidate the profile of lesional and non-lesional keloid skin compared to normal skin. We performed gene (RNAseq, qRT-PCR) and protein (immunohistochemistry) expression analyses on biopsy specimens obtained from lesional and non-lesional skin of African American (AA) keloid patients compared to healthy skin from AA controls. Fold-change≥2 and false-discovery rate (FDR)<0.05 was used to define significance. We found that lesional versus normal skin showed significant up-regulation of markers of T-cell activation/migration (ICOS, CCR7), Th2- (IL-4R, CCL11, TNFSF4/OX40L), Th1- (CXCL9/CXCL10/CXCL11), Th17/Th22- (CCL20, S100As) pathways, and JAK/STAT-signaling (JAK3) (false-discovery rate [FDR]<0.05). Non-lesional skin also exhibited similar trends. We observed increased cellular infiltrates in keloid tissues, including T-cells, dendritic cells, mast cells, as well as greater IL-4rα+, CCR9+, and periostin+ immunostaining. In sum, comprehensive molecular profiling demonstrated that both lesional and non-lesional skin show significant immune alternations, and particularly Th2 and JAK3 expression. This advocates for the investigation of novel treatments targeting the Th2 axis and/or JAK/STAT-signaling in keloid patients.
Cellular and Mathematical Analyses of LUBAC Involvement in T Cell Receptor-Mediated NF-κB Activation Pathway
Authors: Oikawa Daisuke , Hatanaka Naoya , Suzuki Takashi , Tokunaga Fuminori
Published: November 2020

The LUBAC ubiquitin ligase complex, composed of the HOIP, HOIL-1L, and SHARPIN subunits, stimulates the canonical nuclear factor-κB (NF-κB) activation pathways through its Met1-linked linear ubiquitination activity. Here we performed cellular and mathematical modeling analyses of the LUBAC involvement in the T cell receptor (TCR)-mediated NF-κB activation pathway, using the Jurkat human T cell line. LUBAC is indispensable for TCR-induced NF-κB and T cell activation, and transiently associates with and linearly ubiquitinates the CARMA1-BCL10-MALT1 (CBM) complex, through the catalytic HOIP subunit. In contrast, the linear ubiquitination of NEMO, a substrate of the TNF-α-induced canonical NF-κB activation pathway, was limited during the TCR pathway. Among deubiquitinases, OTULIN, but not CYLD, plays a major role in downregulating LUBAC-mediated TCR signaling. Mathematical modeling indicated that linear ubiquitination of the CBM complex accelerates the activation of IκB kinase (IKK), as compared with the activity induced by linear ubiquitination of NEMO alone. Moreover, simulations of the sequential linear ubiquitination of the CBM complex suggested that the allosteric regulation of linear (de)ubiquitination of CBM subunits is controlled by the ubiquitin-linkage lengths. These results indicated that, unlike the TNF-α-induced NF-κB activation pathway, the TCR-mediated NF-κB activation in T lymphocytes has a characteristic mechanism to induce LUBAC-mediated NF-κB activation.
Deep-Sea Coral and Sponge Taxa Increase Demersal Fish Diversity and the Probability of Fish Presence
Authors: Henderson Mark J. , Huff David D. , Yoklavich Mary M.
Published: November 2020

Fishes are known to use deep-sea coral and sponge (DSCS) species as habitat, but it is uncertain whether this relationship is facultative (circumstantial and not restricted to a particular function) or obligate (necessary to sustain fish populations). To explore whether DSCS provide essential habitats for demersal fishes, we analyzed 10 years of submersible survey video transect data, documenting the locations and abundance of DSCS and demersal fishes in the Southern California Bight (SCB). We first classified the different habitats in which fishes and DSCS taxa occurred using cluster analysis, which revealed four distinct DSCS assemblages based on depth and substratum. We then used logistic regression and gradient forest analysis to identify the ecological correlates most associated with the presence of rockfish taxa (Sebastesspp.) and biodiversity. After accounting for spatial autocorrelation, the factors most related to the presence of rockfishes were depth, coral height, and the abundance of a few key DSCS taxa. Of particular interest, we found that young-of-the-year rockfishes were more likely to be present in locations with taller coral and increased densities ofPlumarella longispina,Lophelia pertusa, and two sponge taxa. This suggests these DSCS taxa may serve as important rearing habitat for rockfishes. Similarly, the gradient forest analysis found the most important ecological correlates for fish biodiversity were depth, coral cover, coral height, and a subset of DSCS taxa. Of the 10 top-ranked DSCS taxa in the gradient forest (out of 39 potential DSCS taxa), 6 also were associated with increased probability of fish presence in the logistic regression. The weight of evidence from these multiple analytical methods suggests that this subset of DSCS taxa are important fish habitats. In this paper we describe methods to characterize demersal communities and highlight which DSCS taxa provide habitat to demersal fishes, which is valuable information to fisheries agencies tasked to manage these fishes and their essential habitats.
Hexagon-Twist Frequency Reconfigurable Antennas via Multi-Material Printed Thermo-Responsive Origami Structures
Authors: Zhang Ya-Jing , Wang Li-Chen , Song Wei-Li , Chen Mingji , Fang Daining
Published: November 2020

The origami structure has caused a great interest in the field of engineering, and it has fantastic applications in the deployable and reconfigurable structures. Owing to the unique multi-stable states, here a typical hexagon-twist origami structure is fabricated via multi-material printing technology. The printed structure has multi-stable features and the stiffness of the deformable structure is dramatically reduced under thermal triggering. Such behavior causes an increase in the structural degree of freedom, allowing for self-deployment via releasing the prestored energy in the elastic crease. The response time and reaction time of the self-deployment process are also studied and illustrate the higher energy barrier of the folded state, the longer self-deployment time. Utilizing such unique features and design principles, a prototype of frequency reconfigurable origami antenna of nine diverse operating modes is subsequently designed and assembled with the hexagon-twist origami structure as the dielectric substrate. The antenna implements the cross-band from two different frequency bands, enabling to realize frequency reconfigurable under thermal condition.
X-Linked Alport Syndrome in Women: Genotype and Clinical Course in 24 Cases
Authors: Mastrangelo Antonio , Giani Marisa , Groppali Elena , Castorina Pierangela , Soldà Giulia , Robusto Michela , Fallerini Chiara , Bruttini Mirella , Renieri Alessandra , Montini Giovanni
Published: November 2020

Objectives:X-linked Alport syndrome (XLAS) females are at risk of developing proteinuria and chronic kidney damage (CKD). The aim of this study is to evaluate the genotype-phenotype correlation in this rare population.

Materials and Methods:This is a prospective, observational study of XLAS females, confirmed by a pathogenic mutation inCOL4A5and renal ultrastructural evaluation. Proteinuria, renal function and extrarenal involvement were monitored during follow-up. Patients were divided in 2 groups, according to mutations inCOL4A5: missense (Group 1) and non-missense variants (Group 2).

Results:Twenty-four XLAS females, aged 10.6 ± 10.4 years at clinical onset (mean follow-up: 13.1 ± 12.6 years) were recruited between 2000 and 2017 at a single center. In group 1 there were 10 patients and in group 2, 14 (mean age at the end of follow-up: 24.9 ± 13.6 and 23.2 ± 13.8 years, respectively). One patient in Group 1 and 9 in Group 2 (p= 0.013) developed proteinuria during follow-up. Mean eGFR at last follow-up was lower in Group 2 (p= 0.027), where two patients developed CKD. No differences in hearing loss were documented among the two groups. Two patients in Group 2 carried one mutation in bothCOL4A5andCOL4A3(digenic inheritance) and were proteinuric. In one family, the mother presented only hematuria while the daughter was proteinuric and presented a greater inactivation of the X chromosome carrying the wild-type allele.

Conclusions:The appearance of proteinuria and CKD is more frequent in patients with severe variants. Carrying digenic inheritance and skewed XCI seem to be additional risk factors for proteinuria in XLAS females.
Non-biological Complex Drugs (NBCDs): Complex Pharmaceuticals in Need of Individual Robust Clinical Assessment Before Any Therapeutic Equivalence Decision
Authors: Gaspar Rogério Sá , Silva-Lima Beatriz , Magro Fernando , Alcobia Armando , da Costa Fernando Leal , Feio José
Published: November 2020

Non-Biological Complex Drugs (NBCDs) are complex non-biological drugs comprised of large high molecular weight molecules and, often, nanoparticular structures (including liposomes and block-copolymer micelles). In the case of NBCDs, the entire complex is the active pharmaceutical ingredient and its properties cannot be fully characterized by physicochemical analysis. Moreover, the manufacturing process is fundamental in creating the correct originator product. The same is true for generic versions of the product. A recent appraisal of approval procedures for NBCDs “follow-on products” approved in Europe shows a diversity of regulatory pathways. In fact, three different abridged application procedures, under European legislation, were used: the generic application procedure of Article 10(1), the hybrid application procedure of Article 10(3), and the biosimilar application procedure of Article 10(4). Three informed consent applications via Article 10(c) from innovator companies of glatiramer acetate and sevelamer carbonate were submitted shortly after the approval of the first follow-on products. Furthermore, a number of “well-established use” applications [via Article 10(a)] were approved for iron sucrose and iron dextran complexes. In order to protect patients from the increased risks of NBCD products and NBCD follow-on products, two complementary approaches should be considered: (i) improving the regulatory procedures and their guidance documents within the pre-registration phase, and (ii) not considering interchangeability whenever clinical data is not available. With regards to the latter, the need for adequate safety and efficacy data might also include risk management programmes within post-approval pharmacovigilance actions. This, however, would depend on a risk appraisal that must be considered for individual medicinal products, based on the nature of the submitted relevant set of safety/efficacy data.