bluuebunny/crossref_metadata_2025 huggingface dataset:
The Impact of NOTCH Pathway Alteration on Tumor Microenvironment and Clinical Survival of Immune Checkpoint Inhibitors in NSCLC
Authors: Li Xiaohua , Wang Yuntao , Li Xuebing , Feng Gang , Hu Sheng , Bai Yifeng
Published: July 2021

The treatment of non-small cell lung cancer (NSCLC) with immune checkpoint inhibitors (ICIs) has been proven to induce lasting tumor remission. Screening suitable populations for immunotherapy through predictive markers is an important approach to improving the clinical benefits of patients. Evidence has shown that there may be a close connection between NOTCH signaling and the tumor microenvironment (TME). Hence, we explored the impact of the mutation status of NOTCH signaling on the prognosis of NSCLC patients treated with immunotherapy with the aim to apply NSCLC immunotherapy to the greatest extent possible. We examined two clinical cohorts of NSCLC patients receiving ICIs (MSKCC and NG cohorts). The mutation and prognostic data of the ICI-treated cohort were used to evaluate the association between the mutation status of NOTCH signaling and prognosis following immunotherapy. The expression and mutation data of The Cancer Genome Atlas (TCGA)-NSCLC cohort were used to analyze the differences in the immune microenvironment under different NOTCH signaling mutation states. In the ICI-treated cohorts, the univariate and multivariate Cox regression analyses indicated that high-mutated NOTCH signaling could serve as an independent predictor of NSCLC patients receiving ICIs. Patients with high-mutated NOTCH signaling had significantly improved progression-free survival (PFS) (P = 0.03, HR = 0.69; MSKCC cohort) and prolonged overall survival (OS) (P = 0.004, HR = 0.34; NG cohort). Additionally, high-mutated NOTCH signaling was related to the inflammatory immune microenvironment, inflammatory expression profile, and enhanced immunogenicity. According to this study, high-mutated NOTCH signaling may serve as a biomarker for the prediction of the prognosis of NSCLC patients treated with ICIs. A series of prospective clinical studies and molecular mechanism explorations are still needed in the future.
ILC2s Improve Glucose Metabolism Through the Control of Saturated Fatty Acid Absorption Within Visceral Fat
Authors: Okamura Takuro , Hashimoto Yoshitaka , Mori Jun , Yamaguchi Mihoko , Majima Saori , Senmaru Takafumi , Ushigome Emi , Nakanishi Naoko , Asano Mai , Yamazaki Masahiro , Takakuwa Hiroshi , Satoh Takashi , Akira Shizuo , Hamaguchi Masahide , Fukui Michiaki
Published: July 2021

Background and aims

Group 2 innate lymphoid cells (ILC2s) have been implicated in the regulation of metabolic homeostasis in mice.

Methods

In this study, the role of ILC2s in white adipose tissue (WAT) was investigated using ST2, an IL-33 receptor that is expressed on ILC2 knockout mice.

Results

The deficiency of ST2 decreased ILC2s in WAT, whereas ex-ILC2, which acquired group 1 innate lymphoid cell (ILC1)-like traits, was increased. This led to significant metabolic disorders such as visceral fat obesity, decreased browning in WAT, reduction of energy metabolism, and impaired glucose tolerance, compared to wild type (WT) mice. Those metabolic abnormalities of ST2-knockout (ST2KO) mice were not ameliorated by IL-33 administration, but impaired glucose tolerance and visceral fat obesity were significantly improved by transplantation of ILCs from the bone marrow of WT mice. The relative expression of Cd36 in WAT increased due to the deficiency of ST2, and the storage of saturated fatty acids in WAT of ST2KO mice was significantly higher than that of WT mice. Moreover, saturated fatty acids aggravated the chronic inflammation in adipocytes, promoted the differentiation of M1-like macrophages, and inhibited that of M2-like macrophages.

Conclusions

Our results indicated that ILC2 regulates diet-induced obesity and chronic inflammation through the regulation of saturated fatty acid absorption in visceral adipose tissue.
Allelic Variation in Protein Tyrosine Phosphatase Receptor Type-C in Cattle Influences Erythrocyte, Leukocyte and Humoral Responses to Infestation With the Cattle Tick Rhipicephalus australis
Authors: Jonsson Nicholas N. , Cox David K. , Piper Emily K. , Valdivieso Emily F. Mantilla , Constantinoiu Constantin , Jackson Louise A. , Stear Michael J. , Ross Elizabeth M. , Tabor Ala E.
Published: July 2021

The protein tyrosine phosphatase receptor type-C (PTPRC) gene encodes the common leukocyte antigen (CD45) receptor. CD45 affects cell adhesion, migration, cytokine signalling, cell development, and activation state. Four families of the gene have been identified in cattle: a taurine group (Family 1), two indicine groups (Families 2 and 4) and an African “taurindicine” group (Family 3). Host resistance in cattle to infestation with ticks is moderately heritable and primarily manifests as prevention of attachment and feeding by larvae. This study was conducted to describe the effects of PTPRC genotype on immune-response phenotypes in cattle that display a variable immune responsiveness to ticks. Thirty tick-naïve Santa-Gertrudis cattle (a stabilized composite of 5/8 taurine and 3/8 indicine) were artificially infested with ticks weekly for 13 weeks and ranked according to their tick counts. Blood samples were taken from control and tick-challenged cattle immediately before, then at 21 d after infestation and each subsequent week for 9 weeks. Assays included erythrocyte profiles, white blood cell counts, the percentage of cellular subsets comprising the peripheral blood mononuclear cell (PBMC) population, and the ability of PBMC to recognize and proliferate in response to stimulation with tick antigens in vitro. The cattle were PTPRC genotyped using a RFLP assay that differentiated Family 1 and 3 together (220 bp), from Family 2 (462 bp), and from Family 4 (486 bp). The PTPRC allele frequencies were Family 1/3 = 0.34; Family 2 = 0.47; Family 4 = 0.19. There was no significant association between PTPRC genotype and tick count. Each copy of the Family 1/3 allele significantly decreased total leucocyte count (WCC) and CD8+ cells. Increasing dosage of Family 2 alleles significantly increased red blood cell count (RCC), haematocrit (PCV), and haemoglobin (Hb) concentration in blood. Increasing dosage of the Family 4 allele was associated with increased WCC, reduced RCC, reduced PCV and reduced Hb. Homozygote Family 1/3 animals had consistently lower IgG1 in response to tick Ag than homozygote Family 2 animals. The PTPRC genotype influences the bovine immune response to ticks but was not associated with the observed variation in resistance to tick infestation in this study.
Immune Checkpoint Inhibitors in Human Glioma Microenvironment
Authors: Ghouzlani Amina , Kandoussi Sarah , Tall Mariam , Reddy Konala Priyanka , Rafii Soumaya , Badou Abdallah
Published: July 2021

Gliomas are the most common primary brain tumors in adults. Despite the fact that they are relatively rare, they cause significant morbidity and mortality. High-grade gliomas or glioblastomas are rapidly progressing tumors with a very poor prognosis. The presence of an intrinsic immune system in the central nervous system is now more accepted. During the last decade, there has been no major progress in glioma therapy. The lack of effective treatment for gliomas can be explained by the strategies that cancer cells use to escape the immune system. This being said, immunotherapy, which involves blockade of immune checkpoint inhibitors, has improved patients’ survival in different cancer types. This novel cancer therapy appears to be one of the most promising approaches. In the present study, we will start with a review of the general concept of immune response within the brain and glioma microenvironment. Then, we will try to decipher the role of various immune checkpoint inhibitors within the glioma microenvironment. Finally, we will discuss some promising therapeutic pathways, including immune checkpoint blockade and the body’s effective anti-glioma immune response.
Global Causes, Drivers, and Prevention Measures for Lost Fishing Gear
Authors: Richardson Kelsey , Hardesty Britta Denise , Vince Joanna Zofia , Wilcox Chris
Published: July 2021

Abandoned, Lost or otherwise discarded fishing gear (ALDFG) comprises a significant part of global marine plastic pollution, with adverse consequences for fishers, the seafood industry, and marine wildlife and habitats. To effectively prevent and reduce ALDFG at source, an understanding of the major causes of and drivers behind fishing gear losses is required. We interviewed 451 fishers from seven countries around the world (Belize, Iceland, Indonesia, Morocco, New Zealand, Peru, and the United States of America) representing five key fishing gear types (gillnets, purse seine nets, trawl nets, longlines, and pots and traps) about why and under what circumstances they lose their gear. We also asked them their views on the most effective interventions to reduce gear losses. Across all major gear types and countries where interviews were undertaken, bad weather was the most common cause of gear loss, followed by interactions with wildlife (identified as a cause for loss by 81% and 65% of all fishers interviewed, respectively). Snagging gear on a bottom obstruction was a major cause of loss for gears that contact the seafloor, along with conflicts with other fishers, often via gear and vessel interactions, for gillnet and pot and trap fishers. Operational and behavioral characteristics such as gear type, trip length, and the party responsible to pay for gear repairs and replacements all significantly influenced gear losses. Gear maintenance was the most effective gear loss prevention measure across all gear types and countries reported by fishers, followed by training crew in gear management (identified as an effective prevention measure by 95% and 82% of all fishers interviewed, respectively). Actions available to fishers, managers and port operators to effectively prevent fishing gear losses include: gear maintenance; reducing active gear interactions with wildlife; reducing financial and administrative burdens for port reception facilities; reducing trip lengths; and targeting education and gear stewardship programs to fishers with limited ALDFG awareness, particularly those in low income fisheries and countries.
Influence on Polyurethane Synthesis Parameters Upon the Performance of Base Asphalt
Authors: Jin Xin , Sun Siwei , Guo Naisheng , Huang Shuolei , You Zhanping , Tan Yiqiu
Published: July 2021

The objective of this study aims to investigate the performance of thermoplastic polyurethane elastomers (TPU)-modified asphalt, with special focus on the influence of polyurethane synthesis parameters upon the performance of base asphalt. In this study, the TPU modifier was incorporated into base asphalt to prepare TPU-modified asphalt by using a self-determined laboratory process. The properties of polyester-based TPU and polyether-based TPU modified asphalt with different contents was analyzed by the base performance tests. The Fourier transform infrared spectroscopy (FTIR), fluorescent microscope, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermal gravity (TG), dynamic shear rheology (DSR), and bending beam rheology (BBR) tests were conducted to expose chemical, microstructure, and rheological properties of the asphalt binders, respectively. The results indicated that the modified asphalt with 5% TPU modifier exhibited a favorable performance in terms of the penetration, ductility, softening point, and rotational viscosity, respectively. The isocyanate and polyol reacted to form carbamate in the TPU-modified asphalt, and moreover the isocyanate and aromatic compounds reacted to form halohydrins in the asphalt. Large particles contained in the polyester-based TPU-modified asphalt increased obviously with the increase of Ch (Hard segment content) and r (Isocyanate root index, molar ratio between NCO and OH) value, the fracture mode of it subjected to external load indicated a typical brittle fracture. The content of MDI in the synthesized polyurethane modifier was an important factor affecting the high-temperature stability of the modified asphalt. The polyester-based TPU modifier had a better high temperature performance than the polyether-based TPU modifier. The polyether polyol chain segment showed a greater flexibility as compared to polyester polyol; the addition of polyether polyols can be effectively offset of MDI chain rigidity as r ≤ 1 and Ch < 40% leading to the improvement of the viscoelastic property of asphalt. When Ch = 40% and r = 0.95, the polyether-based TPU-modified asphalt exhibited the same high-temperature grade as the polyester-based TPU-modified asphalt, whereas the polyether-based TPU-modified asphalt showed obviously higher low temperature grades. The polyester-based TPU with Ch = 40% and r = 1 can be used as a modifier to increase the durability of asphalt binders.
Creep Behavior of Near α High Temperature Ti-6.6Al-4.6Sn-4.6Zr-0.9Nb-1.0Mo-0.32Si Alloy
Authors: Yang Dongye , Tian Wenqi , Zhang Xinqi , Si Ke , Li Jiuxiao
Published: July 2021

This study focuses on the microstructure characteristics and tensile and creep properties of a near α high temperature Ti-6.6Al-4.6Sn-4.6Zr-0.9Nb-1.0Mo-0.32Si alloy. Microstructure characteristics were quantitatively investigated using optical microscopy, scanning electron microscope, and transmission electron microscopy. Tensile properties were carried out at room and high temperature. Creep properties were detected under applied stresses ranging from 100–350 MPa at 873–973 K, respectively. Results showed that Widmanstätten microstructure was obtained after hot forged and heat treatment. The strength decreases and the elongation rises with temperature increasing. The ultimate strength and elongation were 1010 MPa, 12% at room temperature, and 620 MPa, 20% at 923 K, respectively. The steady state creep rates rise correspondingly with stress and temperature. Stress exponents are measured within the range of 3.0–3.5. Thus, the creep mechanism is diffusion-controlled viscous glide of dislocation. Ti3Al precipitates are observed. The boundaries and precipitates can obstruct dislocation movement to improve the creep properties. Fracture mechanism of creep is intergranular. The creep mechanism varied from climb of dislocation to sliding of dislocation solution.
MCU-Dependent mROS Generation Regulates Cell Metabolism and Cell Death Modulated by the AMPK/PGC-1α/SIRT3 Signaling Pathway
Authors: Wang Yuxin , Li Xiang , Zhao Fengchao
Published: July 2021

The mitochondrial calcium uniporter is an intensively investigated calcium channel, and its molecular components, structural features, and encoded genes have long been explored. Further studies have shown that the mitochondrial calcium unidirectional transporter (MCU) is a macromolecular complex related to intracellular and extracellular calcium regulation. Based on the current understanding, the MCU is crucial for maintaining cytosolic Ca2+(cCa2+) homeostasis by modulating mitochondrial Ca2+(mCa2+) uptake. The elevation of MCU-induced calcium levels is confirmed to be the main cause of mitochondrial reactive oxygen species (mROS) generation, which leads to disordered cellular metabolic patterns and cell death. In particular, in an I/R injury model, cancer cells, and adipocytes, MCU expression is maintained at high levels. As is well accepted, the AMPK/PGC-1α/SIRT3 pathway is believed to have an affinity for mROS formation and energy consumption. Therefore, we identified a link between MCU-related mROS formation and the AMPK/PGC-1α/SIRT3 signaling pathway in controlling cell metabolism and cell death, which may provide a new possibility of targeting the MCU to reverse relevant diseases.
Adolescent Assent and Reconsent for Biobanking: Recent Developments and Emerging Ethical Issues
Authors: Kasperbauer T. J. , Halverson Colin
Published: July 2021

Research biobanks that enroll minors face important practical, ethical, and regulatory challenges in reconsenting participants when they reach the age of 18. Federal regulations governing research in the United States provide minimal guidance and allow for a range of practices, including waiving the requirement to obtain reconsent. Some commentators have argued that institutional review boards should indeed grant such waivers, given the low risks of biobank-based research and the impracticality of contacting all participants when they turn 18. There is also significant ethical debate about the age at which adolescents can make authentic, autonomous decisions regarding their research participation. This paper reviews these issues in detail, describes the current state of the ethical discussion, and outlines evidence-based policies for enrolling minors into research biobanks.
Interleukin-28B Polymorphisms Predict the Efficacy of Peginterferon Alpha in Patients With Chronic Hepatitis B: A Meta-Analysis
Authors: Ying Sang-Yu , Hu Yao-Ren , Gao Guo-Sheng , Lou Ke-Hong , Huang Zhen
Published: July 2021

Background: Polyethylene glycol interferon alpha (PEG-IFN-α) is the most frequently used pharmacotherapeutic approach in patients infected with hepatitis B virus (HBV). Numerous studies have reported that interleukin-28B (IL-28B) genetic polymorphisms are related to the therapeutic efficacy of PEG-IFN-α, but the results are inconsistent. The present meta-analysis aimed to analyze the association between IL-28B genetic polymorphisms and the prognosis of patients with chronic hepatitis B (CHB) treated with PEG-IFN-α to inform clinical practice.

Methods: PubMed, EBSCO, and Scopus databases were searched for relevant literature published before February 30, 2021. We calculated the crude odds ratios (ORs) with 95% confidence intervals (CIs) of the cited articles. A total of 2510 patients with CHB treated with PEG-IFN-α in 13 clinical cohort studies were analyzed.

Results: The overall analysis demonstrated a potential association between IL-28B genetic polymorphisms and response to PEG-IFN-α; however, the association was not statistically significant. Furthermore, the subgroup analysis revealed that among patients with HBeAg-negative CHB, the rs12979860 CC genotype and rs8099917 TT genotype were associated with more significant treatment response to PEG-IFN-α (CC vs. non-CC: OR 2.78, 95% CI 1.00–7.76, I2 = 83%; TT vs. non-TT: OR 2.16, 95% CI 1.35–3.48, I2 = 0%). Among Asian patients with CHB, the rs12979860 CC genotype was associated with a more significant treatment response to PEG-IFN (CC vs. non-CC: OR 1.88, 95% CI 1.18–2.99, I2 = 0%).

Conclusion: This meta-analysis revealed that the IL-28B rs12979860 CC genotype and rs8099917 TT genotype indicated a better treatment response than non-CC and non-TT genotypes for PEG-IFN-α in patients with CHB.
A Purification Method of 18F-FP-(+)-DTBZ via Solid-Phase Extraction With Combined Cartridges
Authors: Dai Yuyin , Sa Ri , Guan Feng , Wang Qi , Li Yinghua , Zhao Hongguang
Published: July 2021

Background: To optimize [18F] 9-fluoropropyl-(+)-dihydrotetrabenazine (18F-FP-(+)-DTBZ) purification via solid-phase extraction (SPE) with combined cartridges to facilitate its widespread clinical application.

Methods: A modified SPE purification method, employing Sep-Pak PS-2 and Sep-Pak C18 cartridges, was used for the preparation of 18F-FP-(+)-DTBZ. This method was compared to the purification method of high-pressure liquid chromatography (HPLC) and SPE with one cartridge, following quality control test and positron emission tomography (PET) imaging in healthy volunteers and patients with parkinsn's disease (PD).

Results: A SPE purification method integrating Sep-Pak PS-2 and Sep-Pak C18 cartridges was implemented successfully. The retention time of 18F-FP-(+)-DTBZ purified by HPLC, SPE with Sep-Pak PS-2, SPE with Sep-Pak C18, and SPE with combined use of Sep-Pak PS-2 and Sep-Pak C18 cartridges was 8.7, 8.8, 8.7, and 8.9 min, respectively. Fewest impurity peak was detected in 18F-FP-(+)-DTBZ purified by the SPE with combined use of Sep-Pak PS-2 and Sep-Pak C18 cartridges. This modified SPE purification method provided a satisfactory radiochemical yield of 29 ± 1.8% with radiochemical purity >99% and shortened synthesis time to 27 min. The brain uptake of 18F-FP-(+)-DTBZ purified by the modified SPE was comparable to that purified by HPLC in both healthy volunteers and PD patients.

Conclusions: A SPE method integrating Sep-Pak PS-2 and Sep-Pak C18 cartridges for purification of 18F-FP-(+)-DTBZ may be highly suited to automatic synthesis for routine clinical applications, as it provides excellent radiochemical purity, high yield as well as operational simplicity.
ICU-Mortality in Old and Very Old Patients Suffering From Sepsis and Septic Shock
Authors: Bruno Raphael Romano , Wernly Bernhard , Mamandipoor Behrooz , Rezar Richard , Binnebössel Stephan , Baldia Philipp Heinrich , Wolff Georg , Kelm Malte , Guidet Bertrand , De Lange Dylan W. , Dankl Daniel , Koköfer Andreas , Danninger Thomas , Szczeklik Wojciech , Sigal Sviri , van Heerden Peter Vernon , Beil Michael , Fjølner Jesper , Leaver Susannah , Flaatten Hans , Osmani Venet , Jung Christian
Published: July 2021

Purpose: Old (>64 years) and very old (>79 years) intensive care patients with sepsis have a high mortality. In the very old, the value of critical care has been questioned. We aimed to compare the mortality, rates of organ support, and the length of stay in old vs. very old patients with sepsis and septic shock in intensive care.

Methods: This analysis included 9,385 patients, from the multi-center eICU Collaborative Research Database, with sepsis; 6184 were old (aged 65–79 years), and 3,201 were very old patients (aged 80 years and older). A multi-level logistic regression analysis was used to fit three sequential regression models for the binary primary outcome of ICU mortality. A sensitivity analysis in septic shock patients (n = 1054) was also conducted.

Results: In the very old patients, the median length of stay was shorter (50 ± 67 vs. 56 ± 72 h; p < 0.001), and the rate of a prolonged ICU stay was lower (>168 h; 9 vs. 12%; p < 0.001) than the old patients. The mortality from sepsis was higher in very old patients (13 vs. 11%; p = 0.005), and after multi-variable adjustment being very old was associated with higher odds for ICU mortality (aOR 1.32, 95% CI 1.09–1.59; p = 0.004). In patients with septic shock, mortality was also higher in the very old patients (38 vs. 36%; aOR 1.50, 95% CI 1.10–2.06; p = 0.01).

Conclusion: Very old ICU-patients suffer from a slightly higher ICU mortality compared with old ICU-patients. However, despite the statistically significant differences in mortality, the clinical relevance of such minor differences seems to be negligible.
Generation of Infectious Mimivirus Virions Through Inoculation of Viral DNA Within Acanthamoeba castellanii Shows Involvement of Five Proteins, Essentially Uncharacterized
Authors: Sahmi-Bounsiar Dehia , Baudoin Jean-Pierre , Hannat Sihem , Decloquement Philippe , Chabrieres Eric , Aherfi Sarah , La Scola Bernard
Published: July 2021

One of the most curious findings associated with the discovery of Acanthamoeba polyphaga mimivirus (APMV) was the presence of many proteins and RNAs within the virion. Although some hypotheses on their role in Acanthamoeba infection have been put forward, none have been validated. In this study, we directly transfected mimivirus DNA with or without additional proteinase K treatment to extracted DNA into Acanthamoeba castellanii. In this way, it was possible to generate infectious APMV virions, but only without extra proteinase K treatment of extracted DNA. The virus genomes before and after transfection were identical. We searched for the remaining DNA-associated proteins that were digested by proteinase K and could visualize at least five putative proteins. Matrix-assisted laser desorption/ionization time-of-flight and liquid chromatography–mass spectrometry comparison with protein databases allowed the identification of four hypothetical proteins—L442, L724, L829, and R387—and putative GMC-type oxidoreductase R135. We believe that L442 plays a major role in this protein–DNA interaction. In the future, expression in vectors and then diffraction of X-rays by protein crystals could help reveal the exact structure of this protein and its precise role.
Generalist Taxa Shape Fungal Community Structure in Cropping Ecosystems
Authors: Wang Jun-Tao , Shen Ju-Pei , Zhang Li-Mei , Singh Brajesh K. , Delgado-Baquerizo Manuel , Hu Hang-Wei , Han Li-Li , Wei Wen-Xue , Fang Yun-Ting , He Ji-Zheng
Published: July 2021

Fungi regulate nutrient cycling, decomposition, symbiosis, and pathogenicity in cropland soils. However, the relative importance of generalist and specialist taxa in structuring soil fungal community remains largely unresolved. We hypothesized that generalist fungi, which are adaptable to various environmental conditions, could potentially dominate the community and become the basis for fungal coexisting networks in cropping systems. In this study, we identified the generalist and habitat specialist fungi in cropland soils across a 2,200 kms environmental gradient, including three bioclimatic regions (subtropical, warm temperate, and temperate). A few fungal taxa in our database were classified as generalist taxa (~1%). These generalists accounted for >35% of the relative abundance of all fungal populations, and most of them are Ascomycota and potentially pathotrophic. Compared to the specialist taxa (5–17% of all phylotypes in three regions), generalists had a higher degree of connectivity and were often identified as hub within the network. Structural equation modeling provided further evidence that after accounting for spatial and climatic/edaphic factors, generalists had larger contributions to the fungal coexistence pattern than habitat specialists. Taken together, our study provided evidence that generalist taxa are crucial components for fungal community structure. The knowledge of generalists can provide important implication for understanding the ecological preference of fungal groups in cropland systems.
The Genotype (A to H) Dependent N-terminal Sequence of HBV Large Surface Protein Affects Viral Replication, Secretion and Infectivity
Authors: Ou Guomin , He Lingyuan , Wang Luwei , Song Ji , Lai Xinyuan , Tian Xing , Wang Lei , Zhang Kai , Zhang Xuechao , Deng Juan , Zhuang Hui , Xiang Kuanhui , Li Tong
Published: July 2021

Genetic variability has significant impacts on biological characteristics and pathogenicity of hepatitis B virus (HBV), in which the N-terminal sequence of the presurface 1 (preS1) region of HBV large surface protein (LHBs) displays genotype (GT) dependent genetic heterogeneity. However, the influence of this heterogeneity on its biological roles is largely unknown. By analyzing 6560 full-length genome sequences of GTA-GTH downloaded from HBVdb database, the preS1 N-terminal sequences were divided into four representative types, namely C-type (representative of GTA, GTB, and GTC), H-type (GTF and GTH), E-type (GTE and GTG), and D-type (GTD), respectively. We artificially substituted the preS1 N-termini of GTC and GTD plasmids or viral strains with each sequence of the four representative types. The roles of preS1 N-terminus on HBV replication, secretion and infectivity were investigated using HepG2 or HepG2-NTCP cells. In the transfection experiments, the results showed that the extracellular HBsAg levels and HBsAg secretion coefficients in D- and E-type strains were significantly higher than those in C- and H-type strains. D-type strain produced more extracellular HBV DNA than C-type strain. We further observed that D-, H-, and E-type strains increased the levels of intracellular replicative HBV DNAs, comparing with C-type strain. In the infection experiments, the levels of extracellular HBeAg, intracellular HBV total RNA and pgRNA/preC mRNA in D- and E-type strains were markedly higher than C and H-type ones. Our data suggest that the preS1 N-termini affect HBV replication, secretion and infectivity in a genotype dependent manner. The C- and H-type strains prefer to attenuate HBsAg secretion, while the strains of D- and E-type promoted infectivity. The existence and function of the intergenotypic shift of preS1 in naturally occurring recombination requires further investigation, as the data we acquired are mostly related to recombinant preS1 region between N-terminus of preS1 from genotypes A-H and the remaining preS1 portion of GTC or GTD.
Human Cytomegalovirus UL23 Attenuates Signal Transducer and Activator of Transcription 1 Phosphorylation and Type I Interferon Response
Authors: Feng Linyuan , Li Wanwei , Wu Xingyuan , Li Xiaotian , Yang Xiaoping , Ran Yanhong , Wu Jianguo , Li Hongjian
Published: July 2021

Human cytomegalovirus (HCMV), the human beta-herpesvirus, can cause severe syndromes among both immunocompromised adult patients and newborns. Type I interferon (IFN-I) exerts an important effect to resist infections caused by viruses such as HCMV, while IFN evasion may serve as a key determining factor for viral dissemination and disease occurrence within hosts. In this study, UL23, a tegument protein of HCMV, was confirmed to be a key factor for negatively regulating the type I IFN immune response. A detailed analysis indicated that the viral UL23 protein increases the IFN-I antiviral resistance during HCMV infections. Furthermore, UL23 was shown to significantly reduce the levels of IFN-stimulated genes (ISGs) and promoter activity of IFN-I-stimulated response element. Mechanically, UL23 was discovered to impair the signal transducer and activator of transcription 1 (STAT1) phosphorylation, although it was not found to affect phosphorylation and expression of STAT2, Janus activated kinase 1, or tyrosine kinase 2, which are associated with IFN-I signal transduction pathway. Additionally, a significantly reduced nuclear expression of STAT1 but not of IFN regulatory factor 9 or STAT2 was observed. Findings of this study indicate that HCMV UL23 is a viral antagonist that acts against the cellular innate immunity and reveal a possible novel effect of UL23 on IFN-I signaling.
Host- and Species-Dependent Quasispecies Divergence of Severe Acute Respiratory Syndrome Coronavirus-2 in Non-human Primate Models
Authors: Hwang Eun-Ha , Chung Hoyin , Kim Green , Oh Hanseul , An You Jung , Kang Philyong , Ryu Choong-min , Park Jong-Hwan , Hong Jungjoo , Koo Bon-Sang
Published: July 2021

Recently, newly emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been continuously reported worldwide. However, the precise evaluation of SARS-CoV-2 microevolution in host is very limited because the exact genetic information of infected virus could not be acquired in human researches. In this report, we performed deep sequencing for seed virus and SARS-CoV-2 isolated in eight cynomolgus and rhesus macaques at 3 days postinoculation and evaluated single-nucleotide polymorphisms (SNPs) in SARS-CoV-2 by variant analysis. A total of 69 single-nucleotide variants (SNVs) were present in the 5′-untranslated region (UTR), 3′-UTR, ORF1ab, S, ORF3a, ORF8, and N genes of the seed virus passaged in VERO cells. Between those present on the seed virus and those on each SARS-CoV-2 isolated from the lungs of the macaques, a total of 29 variants was identified in 4 coding proteins (ORF1ab, S, ORF3a, and N) and non-coding regions (5′- and 3′-UTR). Variant number was significantly different according to individuals and ranged from 2 to 11. Moreover, the average major frequency variation was identified in six sites between the cynomolgus monkeys and rhesus macaques. As with diverse SNPs in SARS-CoV-2, the values of viral titers in lungs were significantly different according to individuals and species. Our study first revealed that the genomes of SARS-CoV-2 differ according to individuals and species despite infection of the identical virus in non-human primates (NHPs). These results are important for the interpretation of longitudinal studies evaluating the evolution of the SARS-CoV-2 in human beings and development of new diagnostics, vaccine, and therapeutics targeting SARS-CoV-2.
Identification of Microbiota Biomarkers With Orthologous Gene Annotation for Type 2 Diabetes
Authors: Zhang Yu-Hang , Guo Wei , Zeng Tao , Zhang ShiQi , Chen Lei , Gamarra Margarita , Mansour Romany F. , Escorcia-Gutierrez José , Huang Tao , Cai Yu-Dong
Published: July 2021

Type 2 diabetes (T2D) is a systematic chronic metabolic condition with abnormal sugar metabolism dysfunction, and its complications are the most harmful to human beings and may be life-threatening after long-term durations. Considering the high incidence and severity at late stage, researchers have been focusing on the identification of specific biomarkers and potential drug targets for T2D at the genomic, epigenomic, and transcriptomic levels. Microbes participate in the pathogenesis of multiple metabolic diseases including diabetes. However, the related studies are still non-systematic and lack the functional exploration on identified microbes. To fill this gap between gut microbiome and diabetes study, we first introduced eggNOG database and KEGG ORTHOLOGY (KO) database for orthologous (protein/gene) annotation of microbiota. Two datasets with these annotations were employed, which were analyzed by multiple machine-learning models for identifying significant microbiota biomarkers of T2D. The powerful feature selection method, Max-Relevance and Min-Redundancy (mRMR), was first applied to the datasets, resulting in a feature list for each dataset. Then, the list was fed into the incremental feature selection (IFS), incorporating support vector machine (SVM) as the classification algorithm, to extract essential annotations and build efficient classifiers. This study not only revealed potential pathological factors for diabetes at the microbiome level but also provided us new candidates for drug development against diabetes.
Inactivation of Cerebral Cavernous Malformation Genes Results in Accumulation of von Willebrand Factor and Redistribution of Weibel-Palade Bodies in Endothelial Cells
Authors: Much Christiane D. , Sendtner Barbara S. , Schwefel Konrad , Freund Eric , Bekeschus Sander , Otto Oliver , Pagenstecher Axel , Felbor Ute , Rath Matthias , Spiegler Stefanie
Published: July 2021

Cerebral cavernous malformations are slow-flow thrombi-containing vessels induced by two-step inactivation of the CCM1, CCM2 or CCM3 gene within endothelial cells. They predispose to intracerebral bleedings and focal neurological deficits. Our understanding of the cellular and molecular mechanisms that trigger endothelial dysfunction in cavernous malformations is still incomplete. To model both, hereditary and sporadic CCM disease, blood outgrowth endothelial cells (BOECs) with a heterozygous CCM1 germline mutation and immortalized wild-type human umbilical vein endothelial cells were subjected to CRISPR/Cas9-mediated CCM1 gene disruption. CCM1−/− BOECs demonstrated alterations in cell morphology, actin cytoskeleton dynamics, tube formation, and expression of the transcription factors KLF2 and KLF4. Furthermore, high VWF immunoreactivity was observed in CCM1−/− BOECs, in immortalized umbilical vein endothelial cells upon CRISPR/Cas9-induced inactivation of either CCM1, CCM2 or CCM3 as well as in CCM tissue samples of familial cases. Observer-independent high-content imaging revealed a striking reduction of perinuclear Weibel-Palade bodies in unstimulated CCM1−/− BOECs which was observed in CCM1+/− BOECs only after stimulation with PMA or histamine. Our results demonstrate that CRISPR/Cas9 genome editing is a powerful tool to model different aspects of CCM disease in vitro and that CCM1 inactivation induces high-level expression of VWF and redistribution of Weibel-Palade bodies within endothelial cells.
Alteration of N6-Methyladenosine RNA Profiles in Cisplatin-Induced Acute Kidney Injury in Mice
Authors: Li Can-Ming , Li Ming , Zhao Wen-Bo , Ye Zeng-Chun , Peng Hui
Published: July 2021

Aim: To identify the alterations of N6-methyladenosine (m6A) RNA profiles in cisplatin-induced acute kidney injury (Cis-AKI) in mice.

Materials and Methods: The total level of m6A and the expression of methyltransferases and demethylases in the kidneys were measured. The profiles of methylated RNAs were determined by the microarray method. Bioinformatics analysis was performed to predict the functions.

Results: Global m6A levels were increased after cisplatin treatment, accompanied by the alterations of Mettl3, Mettl14, Wtap, Fto, and Alkbh5. A total of 618 mRNAs and 98 lncRNAs were significantly differentially methylated in response to cisplatin treatment. Bioinformatics analysis indicated that the methylated mRNAs predominantly acted on the metabolic process.

Conclusion: M6A epitranscriptome might be significantly altered in Cis-AKI, which is potentially implicated in the development of nephrotoxicity.
Phenotypic and Functional Analyses of B7S1 in Ovarian Cancer
Authors: Cai Dongli , Wang Fang , Wang Changgang , Jin Liping
Published: July 2021

Background: Although programmed death (PD) ligand 1 (PD-L1)/PD-1 inhibitors show potent and durable antitumor effects in a variety of tumors, their efficacy in patients with OvCa is modest. Thus, additional immunosuppressive mechanisms beyond PD-L1/PD-1 need to be identified.

Methods: The mRNA expression profiles of OvCa patients were obtained from The Cancer Genome Atlas (TCGA) database. The expression and clinical characteristics of VTCN1 (encoding B7S1) in OvCa were analyzed. The molecular interaction network, Gene Ontology (GO) analysis and Gene set enrichment analysis (GSEA) were used to functionally annotate and predict signaling pathways of VTCN1 in OvCa. Moreover, 32 treatment-naïve patients with OvCa were recruited to assess B7S1 expression. The cytotoxic immune phenotypes in distinct subgroups were analyzed.

Results: B7S1 expression was increased in tumor sections compared with that in normal tissues from OvCa patients at both the mRNA and protein levels. VTCN1 expression was significantly correlated with the mRNA expression levels of several other co-inhibitory immune checkpoints. B7S1 protein was found to be highly expressed in CD45+CD68+ myeloid cells, whereas its putative receptor was expressed in CD8+ tumor-infiltrating lymphocytes (TILs). Furthermore, expression of B7S1 in antigen-presenting cells (APCs) was significantly correlated with the cytolytic function of CD8+ TILs. Functional annotations indicated that VTCN1 was involved in regulating T cell-mediated immune responses and participated in the activation of a variety of classic signaling pathways related to the progression of human cancer.

Conclusion: In OvCa, B7S1 was highly expressed and may initiate dysfunction of CD8+ TILs, which could be targeted for cancer immunotherapy.
A pH-eQTL Interaction at the RIT2–SYT4 Parkinson’s Disease Risk Locus in the Substantia Nigra
Authors: Patel Sejal , Howard Derek , French Leon
Published: July 2021

Parkinson’s disease causes severe motor and cognitive disabilities that result from the progressive loss of dopamine neurons in the substantia nigra. The rs12456492 variant in the RIT2 gene has been repeatedly associated with increased risk for Parkinson’s disease. From a transcriptomic perspective, a meta-analysis found that RIT2 gene expression is correlated with pH in the human brain. To assess these pH associations in relation to Parkinson’s disease risk, we examined the two datasets that assayed rs12456492, gene expression, and pH in the postmortem human brain. Using the BrainEAC dataset, we replicate the positive correlation between RIT2 gene expression and pH in the human brain (n = 100). Furthermore, we found that the relationship between expression and pH is influenced by rs12456492. When tested across ten brain regions, this interaction is specifically found in the substantia nigra. A similar association was found for the co-localized SYT4 gene. In addition, SYT4 associations are stronger in a combined model with both genes, and the SYT4 interaction appears to be specific to males. In the Genotype-Tissue Expression (GTEx) dataset, the pH associations involving rs12456492 and expression of either SYT4 and RIT2 were not seen. This null finding may be due to the short postmortem intervals of the GTEx tissue samples. In the BrainEAC data, we tested the effect of postmortem interval and only observed the interactions in samples with the longer intervals. These previously unknown associations suggest novel roles for rs12456492, RIT2, and SYT4 in the regulation and response to pH in the substantia nigra.
Influence of Physical Activity Levels and Functional Capacity on Brain β-Amyloid Deposition in Older Women
Authors: Pedrero-Chamizo Raquel , Szoeke Cassandra , Dennerstein Lorraine , Campbell Stephen
Published: July 2021

Physical activity (PA) and Alzheimer's disease are associated. However, how PA influences the cerebral β-amyloid (Aβ) burden remains unclear. The aim of this study was to determine if PA levels and/or functional capacity (FC) are associated with Aβ plaque deposition, and whether these associations differed according to APOE-ε4 genotype. A total of 117 women (69.7 ± 2.6 years; 33.3% APOE-ε4-carriers) from the Women's Healthy Ageing Project cohort (WHAP) were analyzed. PA was measured using the International Physical Activity Questionnaire and, FC was evaluated using the Timed Up and Go test (TUGt). Positron emission tomography with F-18 Florbetaben was carried out to assess cerebral Aβ burden, and quantified using standardized uptake value rations. The sample was split into PA and TUGt tertiles (T1, T2 and T3), and compared according to APOE-ε4 genotype (positive/negative). There were no significant differences in Aβ accumulation according to PA tertiles and APOE-ε4 genotype. Regarding FC, APOE-ε4+ participants in the first TUGt tertile (high performance) obtained significant lower Aβ accumulations compared with the other two tertiles (p < 0.05). Comparing between genotypes, greater Aβ depositions were found between T2 and T3 in APOE-ε4+ compared with those who were APOE-ε4– (p < 0.05). Values of TUGt ≥ 6.5 s (APOE-ε4+) and 8.5 s (APOE-ε4–) were associated with an increased risk of having higher Aβ retention. In conclusion, low performance in TUGt is associated with a negative effect on brain pathology with increasing cerebral Aβ depositions in older women who are APOE-ε4+. In physically active older women (> 600 METs·min/week), higher PA levels are not associated with reduction in Aβ depositions.
Cancer Nanomedicine and Immune System—Interactions and Challenges
Authors: Ahmad Suhana , Idris Ros Akmal Mohd , Wan Hanaffi Wan Nurhidayah , Perumal Komathi , Boer Jennifer C. , Plebanski Magdalena , Jaafar Juhana , Lim Jit Kang , Mohamud Rohimah
Published: July 2021

Nanoparticles have tremendous therapeutic potential in the treatment of cancer as they increase drug delivery, attenuate drug toxicity, and protect drugs from rapid clearance. Since Doxil®, the first FDA-approved nanomedicine, several other cancer nanomedicines have been approved and have successfully increased the efficacy over their free drug counterparts. Although their mechanisms of action are well established, their effects towards our immune system, particularly in the tumor microenvironment (TME), still warrant further investigation. Herein, we review the interactions between an approved cancer nanomedicine with TME immunology. We also discuss the challenges that need to be addressed for the full clinical potential of ongoing cancer nanomedicines despite the encouraging preclinical data.
Impact of Fluoxetine on Behavioral Invigoration of Appetitive and Aversively Motivated Responses: Interaction With Dopamine Depletion
Authors: Carratalá-Ros Carla , López-Cruz Laura , Martínez-Verdú Andrea , Olivares-García Régulo , Salamone John D. , Correa Mercè
Published: July 2021

Impaired behavioral activation and effort-related motivational dysfunctions like fatigue and anergia are debilitating treatment-resistant symptoms of depression. Depressed people show a bias towards the selection of low effort activities. To determine if the broadly used antidepressant fluoxetine can improve behavioral activation and reverse dopamine (DA) depletion-induced anergia, male CD1 mice were evaluated for vigorous escape behaviors in an aversive context (forced swim test, FST), and also with an exercise preference choice task [running wheel (RW)-T-maze choice task]. In the FST, fluoxetine increased active behaviors (swimming, climbing) while reducing passive ones (immobility). However, fluoxetine was not effective at reducing anergia induced by the DA-depleting agent tetrabenazine, further decreasing vigorous climbing and increasing immobility. In the T-maze, fluoxetine alone produced the same pattern of effects as tetrabenazine. Moreover, fluoxetine did not reverse tetrabenazine-induced suppression of RW time but it reduced sucrose intake duration. This pattern of effects produced by fluoxetine in DA-depleted mice was dissimilar from devaluing food reinforcement by pre-feeding or making the food bitter since in both cases sucrose intake time was reduced but animals compensated by increasing time in the RW. Thus, fluoxetine improved escape in an aversive context but decreased relative preference for active reinforcement. Moreover, fluoxetine did not reverse the anergic effects of DA depletion. These results have implications for the use of fluoxetine for treating motivational symptoms such as anergia in depressed patients.
Online Learning and Memory of Neural Trajectory Replays for Prefrontal Persistent and Dynamic Representations in the Irregular Asynchronous State
Authors: Sarazin Matthieu X. B. , Victor Julie , Medernach David , Naudé Jérémie , Delord Bruno
Published: July 2021

In the prefrontal cortex (PFC), higher-order cognitive functions and adaptive flexible behaviors rely on continuous dynamical sequences of spiking activity that constitute neural trajectories in the state space of activity. Neural trajectories subserve diverse representations, from explicit mappings in physical spaces to generalized mappings in the task space, and up to complex abstract transformations such as working memory, decision-making and behavioral planning. Computational models have separately assessed learning and replay of neural trajectories, often using unrealistic learning rules or decoupling simulations for learning from replay. Hence, the question remains open of how neural trajectories are learned, memorized and replayed online, with permanently acting biological plasticity rules. The asynchronous irregular regime characterizing cortical dynamics in awake conditions exerts a major source of disorder that may jeopardize plasticity and replay of locally ordered activity. Here, we show that a recurrent model of local PFC circuitry endowed with realistic synaptic spike timing-dependent plasticity and scaling processes can learn, memorize and replay large-size neural trajectories online under asynchronous irregular dynamics, at regular or fast (sped-up) timescale. Presented trajectories are quickly learned (within seconds) as synaptic engrams in the network, and the model is able to chunk overlapping trajectories presented separately. These trajectory engrams last long-term (dozen hours) and trajectory replays can be triggered over an hour. In turn, we show the conditions under which trajectory engrams and replays preserve asynchronous irregular dynamics in the network. Functionally, spiking activity during trajectory replays at regular timescale accounts for both dynamical coding with temporal tuning in individual neurons, persistent activity at the population level, and large levels of variability consistent with observed cognitive-related PFC dynamics. Together, these results offer a consistent theoretical framework accounting for how neural trajectories can be learned, memorized and replayed in PFC networks circuits to subserve flexible dynamic representations and adaptive behaviors.