bluuebunny/crossref_metadata_2025 huggingface dataset:
Pyrolytic Behavior of Long-Chain Alkyl Quaternary Ammonium Bromide Inside Nanopores
Authors: Niu Yanhui , Yu Wenbin , Yang Shuguang , Wan Quan
Published: December 2021

The pyrolytic behavior of organic matter inside nanopores was studied by simultaneous thermogravimetric/differential scanning calorimetry analyzer coupled with Fourier transform infrared spectroscopy (STA/TG-FTIR). Nanoporous silica was prepared by a hydrothermal method using long-chain alkyl quaternary ammonium bromide (CnTAB, n = 12, 14) as a template. The pyrolytic behavior of CnTAB inside nanopores with different diameters was investigated and compared with that of CnTAB inside and outside nanopores. The results showed that the pyrolytic removal process consisted of the following features: 1) CnTAB underwent carbon chain decomposition and oxidation; 2) the DSC exothermal peak of CnTAB came mainly from its oxidative combustion, and the oxidative combustion temperature decreased with increasing pore size; 3) the CnTAB inside nanopores underwent crystallization–amorphous state phase transition, and CnTAB got trapped inside the calcined nanopores. In addition, the pyrolytic behavior of CnTAB inside the calcined nanopores was found to be similar to that of the uncalcined nanopores. This study aims to understand the storage and transformation processes of organic hydrocarbons under nanopore-confinement effect.
Soil Respiration and Carbon Balance Under Cover Crop in a no-Till Tropical Fruit Orchard
Authors: Freidenreich Ariel , Dattamudi Sanku , Li Yuncong C. , Jayachandran Krishnaswamy
Published: December 2021

Agricultural practices, specifically crop and land management schemes, greatly influence the ability of soil to produce CO2 under varying conditions. A 2-year research study was planned to quantify carbon-dioxide (CO2) emission fluxes and total C (TC) contribution in a no-till tropical soil under carambola with sunn hemp-velvet bean cover cropping (CC) systems. Composted poultry manure (PM) was applied as an additional N source. The treatments were fallow control (F), fallow with PM (FM), sunn hemp (SH), SH with PM (SHM), velvet bean (VB), and VB with PM (VBM). Average daily CO2 emission from VB was 23 and 15% higher than control and SH plots, respectively, during CC growing season. Similarly, CO2 emission after CC termination was highest from VB plots. About 17% higher CO2-C emission was observed from manure applied plots which indicates that additional food sources stimulated microbial activity in the soils and subsequently produced more CO2. However, total C contribution in SH plots were significantly higher than in VB plots and was more apparent when manure was not applied. Soil and air temperature played key roles in CO2 emission, specifically during the CC growing season. Considering both input and output parameters of C in the soil, our results suggest that SH has the better potential in reducing CO2 emission and accumulating more C in the soil than VB in tropical fruit orchard.
Risks for Using FeCl3 Under a Submerged Condition, and Different Water Management to Reduce Uptake of Antimony and Cadmium in a Rice Plant
Authors: Yang JiGang , Wu QianHua , Fan ZhiLian , Feng RenWei
Published: December 2021

Soil pollution by multiple metal(loid)s is a common problem, and it is not easy to synchronously reduce their uptake in crops. Compounds containing iron (Fe) are often used to efficiently remediate soil metal(loid) pollution; however, its associated risks did not receive much attention especially under unsuitable soil water conditions. Pot experiments were set up using an antimony (Sb) and cadmium (Cd) co-contaminated soil treated with a continued submergence condition plus 5, 10, or 20 mg kg−1 FeCl3 (Experiment I), or treated with different water management including submergence, intermittent irrigation, and dry farming (Experiment II). Our results showed that the continued submergence resulted in excessive accumulation of arsenic (As) in different tissues of rice plants even if the soil As background concentration is low. High soil moisture content increased the available concentrations of Sb and As, but reduced that of Cd in rhizosphere soils, which was in line with their concentrations in different tissues of rice plants (Experiment II). Under a continued submergence condition, FeCl3 significantly stimulated As concentration in the shoots, roots (excluded Fe20 treatment), and husks, but reduced it in the grains. FeCl3 reduced Sb concentration only in the roots and grains, and reduced Cd concentration only in the husks, suggesting a limited efficiency of FeCl3 to reduce Cd uptake under a submergence condition. In this study, the dynamic changes of As, Sb, and Cd concentrations in soil solution, their available concentrations in rhizosphere soils, their accumulation in root iron/manganese plaques, and the relationships among the above parameters were also discussed. We suggested that if FeCl3 would be used to remediate the contaminated soils by Sb and Cd, dry farming for a short time is needed to avoid As accumulation, and intermittent irrigation is a potential choice to avoid the excessive accumulation of As, Sb, and Cd in the edible parts of rice plants.
Habitat Specificity Modulates the Response of Small Mammals to Habitat Fragmentation, Loss, and Quality in a Neotropical Savanna
Authors: Mattos Ingrid de , Zimbres Bárbara , Marinho-Filho Jader
Published: December 2021

Landscape conversion of natural environments into agriculture and pasture are driving a marked biodiversity decline in the tropics. Consequences of fragmentation might depend upon habitat amount in the landscape, while the quality of remnants can also affect some species. These factors have been poorly studied in relation to different spatial scales. Furthermore, the impacts of these human-driven alterations may go beyond species loss, possibly causing a loss of ecosystem function and services. In this study, we investigated how changes in landscape configuration (patch size and isolation), habitat loss (considering a landscape gradient of 10, 25, and 40% of remnant forest cover), and habitat quality (forest structure) affect small mammal abundance, richness, taxonomic/functional diversity, and species composition in fragmented landscapes of semideciduous forests in the Brazilian Cerrado. Analyses were performed separately for habitat generalists and forest specialists. We live-trapped small mammals and measured habitat quality descriptors four times in 36 forest patches over the years 2018 and 2019, encompassing both rainy and dry seasons, with a total capture effort of 45,120 trap-nights. Regression analyses indicated that the effect of landscape configuration was not dependent on the proportion of habitat amount in the landscape to determine small mammal assemblages. However, both patch size and habitat loss impacted different aspects of the assemblages in distinct ways. Smaller patches were mainly linked to an overall increase in small mammal abundance, while the abundance of habitat generalists was also negatively affected by habitat amount. Generalist species richness was determined by the proportion of habitat amount in the landscape. Specialist richness was influenced by patch forest quality only, suggesting that species with more demanding habitat requirements might respond to fragmentation and habitat loss at finer scales. Taxonomic or functional diversity were not influenced by landscape structure or habitat quality. However, patch size and habitat amount in the landscape were the major drivers of change in small mammal species composition in semideciduous forests in the Brazilian savanna.
The Cost of Gathering Among the Baka Forager-Horticulturalists From Southeastern Cameroon
Authors: Gallois Sandrine , Henry Amanda G.
Published: December 2021

What present-day foragers do for their living and what they eat have long been privileged areas for exploring human behavior, global health, and human evolution. While many studies have focused on hunting and meat acquisition, less attention has been given to gathering and plant foods. Despite evidence of variation in both nutritional quality and energetic costs of gathering different plants, the overall effort spent on gathering in relation to other subsistence tasks is still under explored. In the current context of economic, climate, and social changes, many forager societies also rely on other subsistence strategies, including agriculture and wage labor. In this study, we aim to explore the place of gathering in the livelihood of a mixed economy society, the Baka forager-horticulturalists of southeastern Cameroon, by comparing the involvement and the costs of activities related to food acquisition. From a pool of 153 adult participants (97 women and 56 men), we collected 246 daily records using a GPS (Global Positioning System) tracker combined with heart rate monitor and time allocation recalls. We compared the duration, distance traveled, and the intensity of work, measured by calculating the metabolic equivalent of task (MET), of subsistence activities related to food acquisition. Results from this work show that gathering activities, performed by both women and men, are energetically costly, with higher MET values than hunting and fishing activities. Furthermore, the MET values vary depending on the targeted plant foods. We discuss these insights in the overall framework of subsistence patterns, merging them with the socio-cultural and environmental factors that might explain Baka livelihood and subsistence strategy.
Common Environmental Pollutants Negatively Affect Development and Regeneration in the Sea Anemone Nematostella vectensis Holobiont
Authors: Klein Sylvia , Frazier Victoria , Readdean Timothy , Lucas Emily , Diaz-Jimenez Erica P. , Sogin Mitchell , Ruff Emil S. , Echeverri Karen
Published: December 2021

The anthozoan sea anemoneNematostella vectensisbelongs to the phylum of cnidarians which also includes jellyfish and corals. Nematostella are native to United States East Coast marsh lands, where they constantly adapt to changes in salinity, temperature, oxygen concentration and pH. Its natural ability to continually acclimate to changing environments coupled with its genetic tractability renderNematostellaa powerful model organism in which to study the effects of common pollutants on the natural development of these animals. Potassium nitrate, commonly used in fertilizers, and Phthalates, a component of plastics are frequent environmental stressors found in coastal and marsh waters. Here we present data showing how early exposure to these pollutants lead to dramatic defects in development of the embryos and eventual mortality possibly due to defects in feeding ability. Additionally, we examined the microbiome of the animals and identified shifts in the microbial community that correlated with the type of water that was used to grow the animals, and with their exposure to pollutants.
Incongruent Spatial Distribution of Taxonomic, Phylogenetic, and Functional Diversity in Neotropical Cocosoid Palms
Authors: Freitas Cintia , Brum Fernanda T. , Cássia-Silva Cibele , Maracahipes Leandro , Carlucci Marcos B. , Collevatti Rosane G. , Bacon Christine D.
Published: December 2021

Biodiversity can be quantified by taxonomic, phylogenetic, and functional diversity. Current evidence points to a lack of congruence between the spatial distribution of these facets due to evolutionary and ecological constraints. A lack of congruence is especially evident between phylogenetic and taxonomic diversity since the name and number of species are an artificial, yet commonly used, way to measure biodiversity. Here we hypothesize that due to evolutionary constraints that link phylogenetic and functional diversity, areas with higher phylogenetic and functional diversity will be spatially congruent in Neotropical cocosoid palms, but neither will be congruent with areas of high taxonomic diversity. Also, we hypothesize that any congruent pattern differs between rainforests and seasonally dry forests, since these palms recently colonized and diversified in seasonally dry ecosystems. We use ecological niche modeling, a phylogenetic tree and a trait database to test the spatial congruence of the three facets of biodiversity. Taxonomic and phylogenetic diversity were negatively correlated. Phylogenetic and functional diversity were positively correlated, even though their spatial congruence was lower than expected at random. Taken together, our results suggest that studies focusing solely on large-scale patterns of taxonomic diversity are missing a wealth of information on diversification potential and ecosystem functioning.
Palladium-Induced Temporal Internalization of MHC Class I Contributes to T Cell-Mediated Antigenicity
Authors: Ito Koyu , Kanaseki Takayuki , Tokita Serina , Torigoe Toshihiko , Hirasawa Noriyasu , Ogasawara Kouetsu
Published: December 2021

Palladium (Pd) is a widely used metal and extremely important biomaterial for the reconstruction of occlusions during dental restorations. However, metallic biomaterials can cause serious allergic reactions, such as Pd-related oral mucositis seen in dentistry. Metal allergy is categorized as a type IV allergy and we demonstrated that CD8 T cells play an important role in Pd allergy previously. As TCR of CD8 T cells recognizes MHC class I/peptide complex, the antigen specificity to this complex seems to be generated during Pd allergy. However, it remains unknown if Pd affects the MHC class I/peptide complex. In this study, we investigated the behavior of the MHC class I/peptide complex in response to Pd treatment. We found that PdCl2 treatment altered peptide presentation on MHC class I and that co-culture with Pd-treated DC2.4 cells induced activation of Pd-responsive TCR-expressing T cell line. Furthermore, PdCl2 treatment induced temporal MHC class I internalization and inhibition of membrane movement suppressed Pd-induced T cell-mediated antigenicity. These data suggest that Pd-induced MHC class I internalization is critical for generation of antigenicity through a mechanism including differential peptide loading on MHC class I, which results in Pd allergy.
IL-6 and TNFα Drive Extensive Proliferation of Human Tregs Without Compromising Their Lineage Stability or Function
Authors: Skartsis Nikolaos , Peng Yani , Ferreira Leonardo M. R. , Nguyen Vinh , Ronin Emilie , Muller Yannick D. , Vincenti Flavio , Tang Qizhi
Published: December 2021

Treg therapies are being tested in clinical trials in transplantation and autoimmune diseases, however, the impact of inflammation on Tregs remains controversial. We challenged human Tregsex-vivowith pro-inflammatory cytokines IL-6 and TNFαand observed greatly enhanced proliferation stimulated by anti-CD3 and anti-CD28 (aCD3/28) beads or CD28 superagonist (CD28SA). The cytokine-exposed Tregs maintained high expression of FOXP3 and HELIOS, demethylated FOXP3 enhancer, and low IFNγ, IL-4, and IL-17 secretion. Blocking TNF receptor using etanercept or deletion ofTNF receptor 2using CRISPR/Cas9 blunted Treg proliferation and attenuated FOXP3 and HELIOS expression. These results prompted us to consider using CD28SA together with IL-6 and TNFαwithout aCD3/28 beads (beadless) as an alternative protocol for therapeutic Treg manufacturing. Metabolomics profiling revealed more active glycolysis and oxidative phosphorylation, increased energy production, and higher antioxidant potential during beadless Treg expansion. Finally, beadless expanded Tregs maintained suppressive functionsin vitroandin vivo. These results demonstrate that human Tregs positively respond to proinflammatory cytokines with enhanced proliferation without compromising their lineage identity or function. This property can be harnessed for therapeutic Treg manufacturing.
A Case Report of Non-Bacterial Cystitis Caused by Immune Checkpoint Inhibitors
Authors: Zhu Sihui , Bian Lijuan , Lv Jia , Liu Baorui , Shen Jie
Published: December 2021

We report a case of non-bacterial cystitis after treatment with programmed death-1 (PD-1) and programmed cell death-ligand 1 (PD-L1) antibodies, which was considered an immune-related adverse event (irAE). A 48-year-old male patient with intrahepatic cholangiocarcinoma (ICC) was treated with nivolumab after postoperative multi-line treatment. This patient recurred worsening of psoriasis and repeated urinary tract discomfort. The drug was discontinued and surgery was performed due to the recurrence of the tumor suggested by imaging. After receiving three cycles of chemotherapy treatment combined with atezolizumab, urinary tract discomfort reappeared. No bacteria were found in multiple urine cultures, and non-bacterial bladder inflammation was considered after cystoscopy biopsy. This is a report of non-bacterial inflammation of the urinary tract caused by immunotherapy.
Intra- and Extracellular Effector Vesicles From Human T And NK Cells: Same-Same, but Different?
Authors: Lettau Marcus , Janssen Ottmar
Published: December 2021

Cytotoxic T lymphocytes (CTL) and Natural Killer (NK) cells utilize an overlapping effector arsenal for the elimination of target cells. It was initially proposed that all cytotoxic effector proteins are stored in lysosome-related effector vesicles (LREV) termed “secretory lysosomes” as a common storage compartment and are only released into the immunological synapse formed between the effector and target cell. The analysis of enriched LREV, however, revealed an uneven distribution of individual effectors in morphologically distinct vesicular entities. Two major populations of LREV were distinguished based on their protein content and signal requirements for degranulation. Light vesicles carrying FasL and 15 kDa granulysin are released in a PKC-dependent and Ca2+-independent manner, whereas dense granules containing perforin, granzymes and 9 kDa granulysin require Ca2+-signaling as a hallmark of classical degranulation. Notably, both types of LREV do not only contain the mentioned cytolytic effectors, but also store and transport diverse other immunomodulatory proteins including MHC class I and II, costimulatory and adhesion molecules, enzymes (i.e. CD26/DPP4) or cytokines. Interestingly, the recent analyses of CTL- or NK cell-derived extracellular vesicles (EV) revealed the presence of a related mixture of proteins in microvesicles or exosomes that in fact resemble fingerprints of the cells of origin. This overlapping protein profile indicates a direct relation of intra- and extracellular vesicles. Since EV potentially also interact with cells at distant sites (apart from the IS), they might act as additional effector vesicles or intercellular communicators in a more systemic fashion.
A Combination of Biomarkers Predict Response to Immune Checkpoint Blockade Therapy in Non-Small Cell Lung Cancer
Authors: Jiang Zedong , Zhou Yao , Huang Juan
Published: December 2021

Immune checkpoint blockade (ICB) therapy has provided clinical benefits for patients with advanced non-small-cell lung cancer (NSCLC), but the majority still do not respond. Although a few biomarkers of ICB treatment response have been developed, the predictive power of these biomarkers showed substantial variation across datasets. Therefore, predicting response to ICB therapy remains a challenge. Here, we provided a concise combinatorial strategy for predicting ICB therapy response and constructed the ICB treatment signature (ITS) in lung cancer. The prediction performance of ITS has been validated in an independent ICB treatment cohort of NSCLC, where patients with higher ITS score were significantly associated with longer progression-free survival and better response. And ITS score was more powerful than traditional biomarkers, such as TMB and PD-L1, in predicting the ICB treatment response in NSCLC. In addition, ITS scores still had predictive effects in other cancer data sets, showing strong scalability and robustness. Further research showed that a high ITS score represented comprehensive immune activation characteristics including activated immune cell infiltration, increased mutation load, and TCR diversity. In conclusion, our practice suggested that the combination of biomarkers will lead to a better prediction of ICB treatment prognosis, and the ITS score will provide NSCLC patients with better ICB treatment decisions.
Dissolved Nitrous Oxide and Hydroxylamine in the South Yellow Sea and the East China Sea During Early Spring: Distribution, Production, and Emissions
Authors: Gu Xueji , Cheng Fang , Chen Xiaolei , Du Guanxiang , Zhang Guiling
Published: December 2021

Coastal marine systems are active regions for the production and emission of nitrous oxide (N2O), a potent greenhouse gas. Due to the inherently high variability in different coastal biogeochemical cycles, the factors and mechanisms regulating coastal N2O cycling remain poorly understood. Hydroxylamine (NH2OH), a potential precursor of N2O, has received less attention than other compounds in the coastal areas. Here, we present the spatial distribution of N2O and the first reported NH2OH distribution in the South Yellow Sea (SYS) and the East China Sea (ECS) between March and April 2017. The surface N2O concentrations in the SYS and the ECS varied from 5.9 to 11.3 nmol L–1 (average of 8.4 ± 1.4 nmol L–1) and were characterized by offshore and north–south decreasing gradients. NH2OH showed patchy characteristics and was highly variable, fluctuating between undetectable to 16.4 nmol L–1. We found no apparent covariation between N2O and NH2OH, suggesting the NH2OH pathway, i.e., nitrification (ammonium oxidation), was not the only process affecting N2O production here. The high NH2OH values co-occurred with the greatest chlorophyll-a and oxygen levels in the nearshore region, along with the relationships between NO2–, NO3–, and NH2OH, indicating that a “fresh” nitrifying system, favoring the production and accumulation of NH2OH, was established during the phytoplankton bloom. The high N2O concentrations were not observed in the nearshore. Based on the correlations of the excess N2O (ΔN2O) and apparent oxygen utilization, as well as ΔN2O vs. NO3–, we concluded that the N2O on the continental shelf was mainly derived from nitrification and nitrifier denitrification. Sea-to-air fluxes of N2O varied from −12.4 to 6.6 μmol m–2 d–1 (−3.8 ± 3.7 μmol m–2 d–1) using the Nightingale et al. (2000) formula and −13.3 to 6.9 μmol m–2 d–1 (−3.9 ± 3.9 μmol m–2 d–1) using the Wanninkhof (2014) formula, which corresponds to 75–112% in saturation, suggesting that the SYS and the ECS acted overall as a sink of atmospheric N2O in early spring, with the strength weakening. Our results reveal the factors and potential mechanisms controlling the production and accumulation of NH2OH and N2O in the SYS and the ECS during early spring.
Seasonal Variability and Future Projection of Ocean Acidification on the East China Sea Shelf off the Changjiang Estuary
Authors: Guo Xianghui , Yao Zhentong , Gao Ying , Luo Yaohua , Xu Yi , Zhai Weidong
Published: December 2021

Ocean acidification (OA) occurs universally in the world’s oceans. Marginal seas are facing more serious OA than the open ocean due to strong anthropogenic and natural impacts. This study investigates carbonate dynamics on the East China Sea (ECS) shelf off the Changjiang Estuary using field observations made from 2015 to 2019 that cover all four seasons. In the low productivity cold seasons, the water was well-mixed vertically. The coastal area and the northern ECS were occupied by water characterized by high dissolved inorganic carbon (DIC), low pH25 (pH at 25°C), and low ΩAr (saturation state index of aragonite), and influenced by the coastal water from the Yellow Sea (YS). However, during highly productive warm seasons, pH25 and ΩAr increased in the surface water but decreased in the bottom water as a result of strong biological DIC uptake in the surface water and CO2 production by strong organic matter remineralization in the bottom water. Strong remineralization decreased pH25 and ΩAr by 0.18 ± 0.08 and 0.73 ± 0.35 in the hypoxic bottom water in summer, even though the bottom water remained oversaturated with respect to aragonite (ΩAr > 1.0) during the surveys. Under the context of global OA and the strong seasonal acidification, the projected bottom water on the ECS shelf will be corrosive for aragonite by mid-century.
Diffuse Large B-Cell Lymphoma of the Mandible Diagnosed by Metagenomic Sequencing: A Case Report
Authors: Liu Kaili , Gao Yang , Han Jiawei , Han Xu , Shi Yingqi , Liu Chao , Li Jie
Published: December 2021

Introduction: Non-Hodgkin lymphoma (NHL) has a much higher incidence rate than Hodgkin lymphoma. Approximately 40% NHL occurs in extranodal tissues or organs, and its clinical manifestations are often nonspecific. Primary bone NHL involving the mandible is an uncommon NHL that is characterized by fever, gum swelling and toothache. Therefore, it is often misdiagnosed as oral diseases.

Case Presentation: A 52-year-old female had recurrent fever for more than 1 month, with numbness in her left jaw and toothache. PET/CT showed an uptake area in the left mandible, suggesting microbial infections. However, antibacterial, and antiviral treatment were ineffective. Furthermore, metagenomic sequencing of plasma reported no pathogens, but instead showed significant copy number variations of multiple chromosomes, which highly suggested the existence of tumor. Finally, diffuse large B-cell lymphoma (DLBCL) was diagnosed by mandibular biopsy, and the patient was transferred to Hematology department for chemotherapy.

Conclusion: mNGS not only assists rapid etiological diagnosis, but also helps rule out infection and diagnose malignant neoplasm.
Locked Nucleic Acid Hydrolysis Probes for the Specific Identification of Probiotic Strains Bifidobacterium animalis subsp. lactis DSM 15954 and Bi-07™
Authors: Shehata Hanan R. , Kiefer Anthony , Morovic Wesley , Newmaster Steven G.
Published: December 2021

Probiotic health benefits are now well-recognized to be strain specific. Probiotic strain characterization and identification is thus important in clinical research and in the probiotic industry. This is becoming especially important with reports of probiotic products failing to meet the declared strain content, potentially compromising their efficacy. Availability of reliable identification methods is essential for strain authentication during discovery, evaluation and commercialization of a probiotic strain. This study aims to develop identification methods for strains Bifidobacterium animalis subsp. lactis DSM 15954 and Bi-07 (Bi-07™) based on real-time PCR, targeting single nucleotide polymorphisms (SNPs). The SNPs were targeted by PCR assays with locked nucleic acid (LNA) probes, which is a novel application in probiotic identification. The assays were then validated following the guidelines for validating qualitative real-time PCR assays. Each assay was evaluated for specificity against 22 non-target strains including closely related Bifidobacterium animalis subsp. lactis strains and were found to achieve 100% true positive and 0% false positive rates. To determine reaction sensitivity and efficiency, three standard curves were established for each strain. Reaction efficiency values were 86, 91, and 90% (R square values > 0.99), and 87, 84, and 86% (R square values > 0.98) for B. animalis subsp. lactis DSM 15954 and Bi-07 assays, respectively. The limit of detection (LOD) was 5.0 picograms and 0.5 picograms of DNA for DSM 15954 and Bi-07 assays, respectively. Each assay was evaluated for accuracy using five samples tested at three different DNA concentrations and both assays proved to be highly repeatable and reproducible. Standard deviation of Cq values between two replicates was always below 1.38 and below 1.68 for DSM 15954 and Bi-07 assays, respectively. The assays proved to be applicable to mono-strain and multi-strain samples as well as for samples in various matrices of foods or dietary supplement ingredients. Overall, the methods demonstrated high specificity, sensitivity, efficiency and precision and broad applicability to sample, matrix and machine types. These methods facilitate strain level identification of the highly monophyletic strains B. animalis subsp. lactis DSM 15954 and Bi-07 to ensure probiotic efficacy and provide a strategy to identify other closely related probiotics organisms.
Glycosylation-on-a-Chip: A Flow-Based Microfluidic System for Cell-Free Glycoprotein Biosynthesis
Authors: Aquino Alicia K. , Manzer Zachary A. , Daniel Susan , DeLisa Matthew P.
Published: December 2021

In recent years, cell-free synthetic glycobiology technologies have emerged that enable production and remodeling of glycoproteins outside the confines of the cell. However, many of these systems combine multiple synthesis steps into one pot where there can be competing reactions and side products that ultimately lead to low yield of the desired product. In this work, we describe a microfluidic platform that integrates cell-free protein synthesis, glycosylation, and purification of a model glycoprotein in separate compartments where each step can be individually optimized. Microfluidics offer advantages such as reaction compartmentalization, tunable residence time, the ability to tether enzymes for reuse, and the potential for continuous manufacturing. Moreover, it affords an opportunity for spatiotemporal control of glycosylation reactions that is difficult to achieve with existing cell-based and cell-free glycosylation systems. In this work, we demonstrate a flow-based glycoprotein synthesis system that promotes enhanced cell-free protein synthesis, efficient protein glycosylation with an immobilized oligosaccharyltransferase, and enrichment of the protein product from cell-free lysate. Overall, this work represents a first-in-kind glycosylation-on-a-chip prototype that could find use as a laboratory tool for mechanistic dissection of the protein glycosylation process as well as a biomanufacturing platform for small batch, decentralized glycoprotein production.
Reactive Astrocytes in Central Nervous System Injury: Subgroup and Potential Therapy
Authors: Yu GuiLian , Zhang Ying , Ning Bin
Published: December 2021

Traumatic central nervous system (CNS) injury, which includes both traumatic brain injury (TBI) and spinal cord injury (SCI), is associated with irreversible loss of neurological function and high medical care costs. Currently, no effective treatment exists to improve the prognosis of patients. Astrocytes comprise the largest population of glial cells in the CNS and, with the advancements in the field of neurology, are increasingly recognized as having key functions in both the brain and the spinal cord. When stimulated by disease or injury, astrocytes become activated and undergo a series of changes, including alterations in gene expression, hypertrophy, the loss of inherent functions, and the acquisition of new ones. Studies have shown that astrocytes are highly heterogeneous with respect to their gene expression profiles, and this heterogeneity accounts for their observed context-dependent phenotypic diversity. In the inured CNS, activated astrocytes play a dual role both as regulators of neuroinflammation and in scar formation. Identifying the subpopulations of reactive astrocytes that exert beneficial or harmful effects will aid in deciphering the pathological mechanisms underlying CNS injuries and ultimately provide a theoretical basis for the development of effective strategies for the treatment of associated conditions. Following CNS injury, as the disease progresses, astrocyte phenotypes undergo continuous changes. Although current research methods do not allow a comprehensive and accurate classification of astrocyte subpopulations in complex pathological contexts, they can nonetheless aid in understanding the roles of astrocytes in disease. In this review, after a brief introduction to the pathology of CNS injury, we summarize current knowledge regarding astrocyte activation following CNS injury, including: (a) the regulatory factors involved in this process; (b) the functions of different astrocyte subgroups based on the existing classification of astrocytes; and (c) attempts at astrocyte-targeted therapy.
A Preliminary DTI Tractography Study of Developmental Neuroplasticity 5–15 Years After Early Childhood Traumatic Brain Injury
Authors: Wilde Elisabeth A. , Hyseni Ilirjana , Lindsey Hannah M. , Faber Jessica , McHenry James M. , Bigler Erin D. , Biekman Brian D. , Hollowell Laura L. , McCauley Stephen R. , Hunter Jill V. , Ewing-Cobbs Linda , Aitken Mary E. , MacLeod Marianne , Chu Zili D. , Noble-Haeusslein Linda J. , Levin Harvey S.
Published: December 2021

Plasticity is often implicated as a reparative mechanism when addressing structural and functional brain development in young children following traumatic brain injury (TBI); however, conventional imaging methods may not capture the complexities of post-trauma development. The present study examined the cingulum bundles and perforant pathways using diffusion tensor imaging (DTI) in 21 children and adolescents (ages 10–18 years) 5–15 years after sustaining early childhood TBI in comparison with 19 demographically-matched typically-developing children. Verbal memory and executive functioning were also evaluated and analyzed in relation to DTI metrics. Beyond the expected direction of quantitative DTI metrics in the TBI group, we also found qualitative differences in the streamline density of both pathways generated from DTI tractography in over half of those with early TBI. These children exhibited hypertrophic cingulum bundles relative to the comparison group, and the number of tract streamlines negatively correlated with age at injury, particularly in the late-developing anterior regions of the cingulum; however, streamline density did not relate to executive functioning. Although streamline density of the perforant pathway was not related to age at injury, streamline density of the left perforant pathway was significantly and positively related to verbal memory scores in those with TBI, and a moderate effect size was found in the right hemisphere. DTI tractography may provide insight into developmental plasticity in children post-injury. While traditional DTI metrics demonstrate expected relations to cognitive performance in group-based analyses, altered growth is reflected in the white matter structures themselves in some children several years post-injury. Whether this plasticity is adaptive or maladaptive, and whether the alterations are structure-specific, warrants further investigation.
Unmet Needs for Rehabilitative Management in Common Health-Related Problems Negatively Impact the Quality of Life of Community-Dwelling Stroke Survivors
Authors: Kim Kyoung Tae , Chang Won Kee , Jung Yun-Sun , Jee Sungju , Sohn Min Kyun , Ko Sung-Hwa , Shin Yong-Il , Leigh Ja-Ho , Kim Won-Seok , Paik Nam-Jong
Published: December 2021

Purpose: Community-dwelling stroke survivors have various unmet needs for rehabilitative management, but there is a lack of in-depth investigations on common health problems after stroke. Moreover, the association between unmet needs and health-related quality of life (HRQoL) has not been thoroughly investigated. This study aimed to investigate the unmet needs for rehabilitative management in common problems after stroke and their associations with HRQoL among community-dwelling stroke survivors.

Methods: A face-to-face cross-sectional survey was conducted among community-dwelling stroke survivors visiting outpatient clinics of rehabilitation departments between June and October 2020 in three university-affiliated hospitals. Unmet needs for common problems after stroke were assessed across eight domains based on the post-stroke checklist: spasticity, dysphagia, communication, cognition, ambulation, pain/discomfort, anxiety/depression, and self-care. HRQoL was measured using the EuroQoL-5D three level (EQ-5D). The prevalence of unmet needs for rehabilitative management and their associations with the EQ-5D index were analyzed.

Results: Among the 239 participants who responded to the survey, 63% (n = 150) were men. The mean age was 63 ± 13 years, and the mean duration of stroke onset was 55.6 months. Overall, 49% reported at least one unmet need, and the most frequently reported unmet needs were anxiety/depression (20.9%), self-care (20.9%), and pain/discomfort (18.0%). The highest proportion of unmet needs was in the anxiety/depression, communication, and cognition domains. Patients with unmet needs for cognition and pain/discomfort showed a significantly lower EQ-5D index, even after adjusting for age, sex, and modified Rankin scale scores. The total number of unmet needs was significantly correlated with a lower EQ-5D index (Pearson's r = −0.329, p < 0.001) in the multivariate linear regression model.

Conclusions: Unmet rehabilitative needs are prevalent among community-dwelling stroke survivors, and the proportion of unmet needs was high among non-physical domains such as anxiety/depression. The number of unmet needs is an independent negative predictor of HRQoL. Systematic approaches to identify unmet needs and provide appropriate rehabilitative management are required in long-term stroke survivors.
Evaluation of Serum Levels of Transient Receptor Potential Cation Channel Subfamily V Member 1, Vasoactive Intestinal Polypeptide, and Pituitary Adenylate Cyclase-Activating Polypeptide in Chronic and Episodic Migraine: The Possible Role in Migraine Transformation
Authors: Togha Mansoureh , Ghorbani Zeinab , Ramazi Samira , Zavvari Fahime , Karimzadeh Fariba
Published: December 2021

Objectives: This study aimed to investigate the role of serum levels of transient receptor potential cation channel subfamily V member 1 (TRPV1), vasoacive intestinal peptide (VIP), and pituitary adenylate cyclase-activating polypeptide (PACAP) in the development and also the transformation of migraine in patients suffering from migraine.

Methods: Eighty-nine participants with a mean age of 39 years were divided into 23 episodic migraine (EM), 36 chronic migraine (CM), and 30 healthy control groups. Demographic, anthropometric, and headache characteristic information, and also blood samples, was collected. Serum levels of TRPV1, VIP, and PACAP were measured using the enzyme-linked immunosorbent assay (ELISA) technique.

Results: Based on our findings, the serum level of TRPV1 was significantly higher in CM compared to the control group (p < 0.05), whereas serum levels of VIP (p < 0.01) and PACAP (p < 0.05) in the EM group were significantly more than the control group. There was no significant difference between EM and CM groups.

Conclusions: An elevation in the serum levels of TRVP1 among chronic migraineurs and increments in the levels of VIP and PACAP were observed among EM patients compared to healthy subjects. However, our data failed to demonstrate the probable role of these biomarkers in migraine progression, and more studies are needed to clarify the molecular mechanisms involved in migraine progression.
B-Mode Ultrasound, a Reliable Tool for Monitoring Experimental Intracerebral Hemorrhage
Authors: Gómez-de Frutos Mari Carmen , García-Suárez Iván , Laso-García Fernando , Diekhorst Luke , Otero-Ortega Laura , Alonso de Leciñana María , Fuentes Blanca , Gutiérrez-Fernández María , Díez-Tejedor Exuperio , Ruíz-Ares Gerardo
Published: December 2021

Background: Magnetic resonance imaging (MRI) is currently used for the study of intracerebral hemorrhage (ICH) in animal models. However, ultrasound is an inexpensive, non-invasive and rapid technique that could facilitate the diagnosis and follow-up of ICH. This study aimed to evaluate the feasibility and reliability of B-mode ultrasound as an alternative tool for in vivo monitoring of ICH volume and brain structure displacement in an animal model.

Methods: A total of 31 male and female Sprague-Dawley rats were subjected to an ICH model using collagenase-IV in the striatum following stereotaxic references. The animals were randomly allocated into 3 groups: healthy (n = 10), sham (n = 10) and ICH (n = 11). B-mode ultrasound studies with a 13-MHz probe were performed pre-ICH and at 5 h, 48 h, 4 d and 1 mo post-ICH for the assessment of ICH volume and displacement of brain structures, considering the distance between the subarachnoid cisterns and the dura mater. The same variables were studied by MRI at 48 h and 1 mo post-ICH.

Results: Both imaging techniques showed excellent correlation in measuring ICH volume at 48 h (r = 0.905) and good at 1 mo (r = 0.656). An excellent correlation was also observed in the measured distance between the subarachnoid cisterns and the dura mater at 1 mo between B-mode ultrasound and MRI, on both the ipsilateral (r = 0.870) and contralateral (r = 0.906) sides of the lesion.

Conclusion: B-mode ultrasound imaging appears to be a reliable tool for in vivo assessment of ICH volume and displacement of brain structures in animal models.
Differentiating Progressive Supranuclear Palsy and Parkinson's Disease With Head-Mounted Displays
Authors: Herwig Arvid , Agic Almedin , Huppertz Hans-Jürgen , Klingebiel Randolf , Zuhorn Frédéric , Schneider Werner X. , Schäbitz Wolf-Rüdiger , Rogalewski Andreas
Published: December 2021

Background: Progressive supranuclear palsy (PSP) is a neurodegenerative disorder that, especially in the early stages of the disease, is clinically difficult to distinguish from Parkinson's disease (PD).

Objective: This study aimed at assessing the use of eye-tracking in head-mounted displays (HMDs) for differentiating PSP and PD.

Methods: Saccadic eye movements of 13 patients with PSP, 15 patients with PD, and a group of 16 healthy controls (HCs) were measured. To improve applicability in an inpatient setting and standardize the diagnosis, all the tests were conducted in a HMD. In addition, patients underwent atlas-based volumetric analysis of various brain regions based on high-resolution MRI.

Results: Patients with PSP displayed unique abnormalities in vertical saccade velocity and saccade gain, while horizontal saccades were less affected. A novel diagnostic index was derived, multiplying the ratios of vertical to horizontal gain and velocity, allowing segregation of PSP from PD with high sensitivity (10/13, 77%) and specificity (14/15, 93%). As expected, patients with PSP as compared with patients with PD showed regional atrophy in midbrain volume, the midbrain plane, and the midbrain tegmentum plane. In addition, we found for the first time that oculomotor measures (vertical gain, velocity, and the diagnostic index) were correlated significantly to midbrain volume in the PSP group.

Conclusions: Assessing eye movements in a HMD provides an easy to apply and highly standardized tool to differentiate PSP of patients from PD and HCs, which will aid in the diagnosis of PSP.
A Deletion of the Nuclear Localization Signal Domain in the Fus Protein Induces Stable Post-stress Cytoplasmic Inclusions in SH-SY5Y Cells
Authors: Notaro Antonietta , Messina Antonella , La Bella Vincenzo
Published: December 2021

Mutations in Fused-in-Sarcoma (FUS) gene involving the nuclear localization signal (NLS) domain lead to juvenile-onset Amyotrophic Lateral Sclerosis (ALS). The mutant protein mislocalizes to the cytoplasm, incorporating it into Stress Granules (SG). Whether SGs are the first step to the formation of stable FUS-containing aggregates is still unclear. In this work, we used acute and chronic stress paradigms to study the SG dynamics in a human SH-SY5Y neuroblastoma cell line carrying a deletion of the NLS domain of the FUS protein (homozygous: ΔNLS–/–; heterozygous: ΔNLS+/–). Wild-type (WT) cells served as controls. We evaluated the subcellular localization of the mutant protein through immunoblot and immunofluorescence, in basal conditions and after acute stress and chronic stress with sodium arsenite (NaAsO2). Cells were monitored for up to 24 h after rescue. FUS was expressed in both nucleus and cytoplasm in the ΔNLS+/– cells, whereas it was primarily cytoplasmic in the ΔNLS–/–. Acute NaAsO2 exposure induced SGs: at rescue,>90% of ΔNLS cells showed abundant FUS-containing if compared to less than 5% of the WT cells. The proportion of FUS-positive SGs remained 15–20% at 24 h in mutant cells. Cycloheximide did not abolish the long-lasting SGs in mutant cells. Chronic exposure to NaAsO2 did not induce significant SGs formation. A wealth of research has demonstrated that ALS-associated FUS mutations at the C-terminus facilitate the incorporation of the mutant protein into SGs. We have shown here that mutant FUS-containing SGs tend to fail to dissolve after stress, facilitating a liquid-to-solid phase transition. The FUS-containing inclusions seen in the dying motor neurons might therefore directly derive from SGs. This might represent an attractive target for future innovative therapies.
Advances With Non-coding RNAs in Neuropathic Pain
Authors: Hu Cheng , He Menglin , Xu Qian , Tian Weiqian
Published: December 2021

Neuropathic pain (NP) is one of the most common types of clinical pain. The common causes of this syndrome include injury to the central or peripheral nervous systems and pathological changes. NP is characterized by spontaneous pain, hyperalgesia, abnormal pain, and paresthesia. Because of its diverse etiology, the pathogenesis of NP has not been fully elucidated and has become one of the most challenging problems in clinical medicine. This kind of pain is extremely resistant to conventional treatment and is accompanied by serious complications. Non-coding RNAs (ncRNAs), such as microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs), contribute to diverse biological processes by regulating the expression of various mRNAs involved in pain-related pathways, at the posttranscriptional level. Abnormal regulation of ncRNAs is closely related to the occurrence and development of NP. In this review, we summarize the current state of understanding of the roles of different ncRNAs in the development of NP. Understanding these mechanisms can help develop novel therapeutic strategies to prevent or treat chronic pain.
Autism-Like Behavior in the Offspring of CYP11A1-Overexpressing Pregnant Rats
Authors: Pan Tianying , Jiang Chuan , Cheng Juan , Xie Jiang , Liu Xinghui , Xu Wenming , He Guolin
Published: December 2021

Autism spectrum disorders (ASD) represent a complex group of neurodevelopmental disorders that are characterized by impaired social behavior and communication as well as repetitive behavior and restricted interests. Prenatal exposure to high levels of testosterone and preeclampsia are thought to be risk factors of ASD. We had previously reported that overexpression of the mitochondrial cholesterol side-chain cleavage enzyme (CYP11A1) could lead to both preeclampsia-like symptoms and increased testosterone levels in pregnant rats. In this study, we investigated the association between high CYP11A1 levels in pregnant rats and autism-like behavior in their offspring. Timed-pregnant Sprague-Dawley (SD) rats were injected with CYP11A1 gene-carrying adenoviruses on gestational day 8.5, and their offspring were then compared with those from timed-pregnant control SD rats. Compared with their control counterparts, the offspring of the CYP11A1-ovexpressing dams displayed more symptoms of anxiety and spent less time in social interactions and more time in self-grooming and rearing, all indicators of autism-like behavior. Sequencing of the transcriptome in primary microglia from the offspring of CYP11A1-overexpressing dams revealed that immune pathways were highly activated, and the gamma-aminobutyric acid type A (GABAA) receptor genes were among the top differentially expressed genes. Using primary microglia cultures generated from neonatal rats, tumor necrosis factor-alpha expression was found to be elevated in the cells transfected with CYP11A1-carrying adenoviruses. Additionally, the offspring of CYP11A1-overexpressing dams displayed dysregulated GABAA receptor expression. Taken together, these results suggest that abnormal CYP11A1 gene expression in pregnant rats could lead to microglial immune activation and dysregulated GABAA receptor expression in their offspring and thereby anxiety and autism-related behavior. Our study suggests that the pathways regulated by CYP11A1 could be promising preventative and therapeutic targets for ASD.