bluuebunny/crossref_metadata_2025 huggingface dataset:
Circadian Rhythm Disruption Influenced Hepatic Lipid Metabolism, Gut Microbiota and Promoted Cholesterol Gallstone Formation in Mice
Authors: He Chuanqi , Shen Weiyi , Chen Chaobo , Wang Qihan , Lu Qifan , Shao Wentao , Jiang Zhaoyan , Hu Hai
Published: October 2021

Background

Hepatic lipid metabolism regulates biliary composition and influences the formation of cholesterol gallstones. The genes Hmgcr and Cyp7a1, which encode key liver enzymes, are regulated by circadian rhythm-related transcription factors. We aimed to investigate the effect of circadian rhythm disruption on hepatic cholesterol and bile acid metabolism and the incidence of cholesterol stone formation.

Methods

Adult male C57BL/6J mice were fed either a lithogenic diet (LD) only during the sleep phase (time-restricted lithogenic diet feeding, TRF) or an LD ad libitum (non-time-restricted lithogenic diet feeding, nTRF) for 4 weeks. Food consumption, body mass gain, and the incidence of gallstones were assessed. Circulating metabolic parameters, lipid accumulation in the liver, the circadian expression of hepatic clock and metabolic genes, and the gut microbiota were analyzed.

Results

TRF caused a dysregulation of the circadian rhythm in the mice, characterized by significant differences in the circadian expression patterns of clock-related genes. In TRF mice, the circadian rhythms in the expression of genes involved in bile acid and cholesterol metabolism were disrupted, as was the circadian rhythm of the gut microbiota. These changes were associated with high biliary cholesterol content, which promoted gallstone formation in the TRF mice.

Conclusion

Disordered circadian rhythm is associated with abnormal hepatic bile acid and cholesterol metabolism in mice, which promotes gallstone formation.
The Relevant Participation of Prolactin in the Genesis and Progression of Gynecological Cancers
Authors: Ramírez-de-Arellano Adrián , Villegas-Pineda Julio César , Hernández-Silva Christian David , Pereira-Suárez Ana Laura
Published: October 2021

Prolactin (PRL) is a hormone produced by the pituitary gland and multiple non-pituitary sites, vital in several physiological processes such as lactation, pregnancy, cell growth, and differentiation. However, PRL is nowadays known to have a strong implication in oncogenic processes, making it essential to delve into the mechanisms governing these actions. PRL and its receptor (PRLR) activate a series of effects such as survival, cellular proliferation, migration, invasion, metastasis, and resistance to treatment, being highly relevant in developing certain types of cancer. Because women produce high levels of PRL, its influence in gynecological cancers is herein reviewed. It is interesting that, other than the 23 kDa PRL, whose mechanism of action is endocrine, other variants of PRL have been observed to be produced by tumoral tissue, acting in a paracrine/autocrine manner. Because many components, including PRL, surround the microenvironment, it is interesting to understand the hormone’s modulation in cancer cells. This work aims to review the most important findings regarding the PRL/PRLR axis in cervical, ovarian, and endometrial cancers and its molecular mechanisms to support carcinogenesis.
Air Warming and Drainage Influences Soil Microarthropod Communities
Authors: Zhang Hui , Sun Xin , Liu Dong , Wu Haitao , Chen Huai
Published: October 2021

The degradation of wetlands due to climate change is of critical concern to human beings worldwide. Little is known about the potential synergistic effects of simultaneous water level reduction and warming on the underground wetland ecosystems. We conducted a 5-month field experiment in the Sanjiang Plain, utilizing open-top chambers and water level automatic control systems to investigate such synergistic effects. Soil springtails (Collembola) and mites (Acari) in the top (0–20 cm) soil layers were sampled to calculate their density, diversity, and to screen for indicator species. Warming significantly influenced soil springtail communities, slightly increasing the total density and total abundance under the natural water level while reducing them under a constant water level. In addition, Anurida maritima and Vertagopus laricis, two indicators for the natural water level, had the highest densities in the natural water level treatment and under the combined treatment of warming and natural water level, respectively. Cheiroseius sinicus and Malaconothrus tardus had the highest densities in warming under the 0 cm water level, significantly higher than the other three treatments. This study also revealed the importance of maintaining fluctuating water levels for microarthropod communities influenced by global warming, providing a theoretical basis for water level control in wetland restoration.
Identification of a Ferroptosis-Related Long Noncoding RNA Prognostic Signature and Its Predictive Ability to Immunotherapy in Hepatocellular Carcinoma
Authors: Wang Liang , Ge Xiangwei , Zhang Zifeng , Ye Yating , Zhou Ziyi , Li Manhong , Yan Hongxiang , Wu Lei , Bai Qian , Li Jipeng , Zhu Jun , Wang Yusheng
Published: October 2021

Background: Immune checkpoint blockers (ICBs) are increasingly being used to treat patients with advanced hepatocellular carcinoma (HCC), but only a third of these patients are sensitive to ICBs. Emerging evidence suggests that ferroptosis could be a novel target for antitumor treatment, and combined treatment with ferroptosis inducers might enhance sensitivity to immunotherapy. However, there is a lack of information on the crosstalk between ferroptosis-related lncRNAs and anti-tumor immunity. Therefore, we aim to explore prognostic value of ferroptosis-related lncRNAs and clarify potential role in ICBs of HCC.

Methods: We obtained mRNA and lncRNA expression data from two independent cohorts (TCGA and GEO database). Univariate Cox, the least absolute shrinkage and selection operator (Lasso) algorithm and multivariate Cox analysis were used to construct a lncRNA signature, which was evaluated using the area under the receiver operating characteristic curve (AUC) and Kaplan–Meier curves. Tumor-infiltrating cell (TIC) profiling and the tumor immune dysfunction and exclusion (TIDE) algorithm were used to validate the signature model and immunotherapy. Finally, we adopted RT-PCR assay to evaluate the differential expression of lncRNAs in HCC tissues in our hospital.

Results: The ferroptosis-related lncRNA signature included five lncRNAs, most of which were positively correlated with clinical stage and grade. The signature could stratify patients into two risk groups, with the high-risk group associated with a shorter overall survival (OS, p < 0.05) in TCGA-LIHC and GSE76427. Besides, the AUCs of the 1-, 3-, and 5-years OS were 0.772, 0.707, and 0.666, respectively. Gene set enrichment analysis (GESA) of lncRNAs revealed enrichment of oncogenic and immune-related pathways. The TIC profiling indicated a close correlation between the signature and immune cells. Furthermore, the high-risk group had a better response to immunotherapy than low-risk group. RT-PCR demonstrated these five lncRNAs were upregulated in cancerous tissue than normal tissues.

Conclusions: The ferroptosis-related lncRNA signature could accurately predict the OS of HCC patients and may serve as an independent clinical factor for patients’ outcomes. Ferroptosis-related lncRNAs may remodel the tumor microenvironment (TME) and affect the anti-cancer ability of ICBs, and therefore, could potentially act as an indicator for the response to immunotherapy in HCC.
Meta-Analysis of Esophageal Cancer Transcriptomes Using Independent Component Analysis
Authors: Ashenova Ainur , Daniyarov Asset , Molkenov Askhat , Sharip Aigul , Zinovyev Andrei , Kairov Ulykbek
Published: October 2021

Independent Component Analysis is a matrix factorization method for data dimension reduction. ICA has been widely applied for the analysis of transcriptomic data for blind separation of biological, environmental, and technical factors affecting gene expression. The study aimed to analyze the publicly available esophageal cancer data using the ICA for identification and comprehensive analysis of reproducible signaling pathways and molecular signatures involved in this cancer type. In this study, four independent esophageal cancer transcriptomic datasets from GEO databases were used. A bioinformatics tool « BiODICA—Independent Component Analysis of Big Omics Data» was applied to compute independent components (ICs). Gene Set Enrichment Analysis (GSEA) and ToppGene uncovered the most significantly enriched pathways. Construction and visualization of gene networks and graphs were performed using the Cytoscape, and HPRD database. The correlation graph between decompositions into 30 ICs was built with absolute correlation values exceeding 0.3. Clusters of components—pseudocliques were observed in the structure of the correlation graph. The top 1,000 most contributing genes of each ICs in the pseudocliques were mapped to the PPI network to construct associated signaling pathways. Some cliques were composed of densely interconnected nodes and included components common to most cancer types (such as cell cycle and extracellular matrix signals), while others were specific to EC. The results of this investigation may reveal potential biomarkers of esophageal carcinogenesis, functional subsystems dysregulated in the tumor cells, and be helpful in predicting the early development of a tumor.
Validation of Reference Genes for Quantitative Real-Time PCR Normalization in Ananas comosus var. bracteatus During Chimeric Leaf Development and Response to Hormone Stimuli
Authors: Mao Meiqin , Xue Yanbin , He Yehua , Zhou Xuzixing , Hu Hao , Liu Jiawen , Feng Lijun , Yang Wei , Luo Jiaheng , Zhang Huiling , Li Xi , Ma Jun
Published: October 2021

Reverse transcription quantitative real-time PCR (RT-qPCR) is a common way to study gene regulation at the transcriptional level due to its sensibility and specificity, but it needs appropriate reference genes to normalize data. Ananas comosus var. bracteatus, with white-green chimeric leaves, is an important pantropical ornamental plant. Up to date, no reference genes have been evaluated in Ananas comosus var. bracteatus. In this work, we used five common statistics tools (geNorm, NormFinder, BestKeeper, ΔCt method, RefFinder) to evaluate 10 candidate reference genes. The results showed that Unigene.16454 and Unigene.16459 were the optimal reference genes for different tissues, Unigene.16454 and zinc finger ran-binding domain-containing protein 2 (ZRANB2) for chimeric leaf at different developmental stages, isocitrate dehydrogenase [NADP] (IDH) and triacylglycerol lipase SDP1-like (SDP) for seedlings under different hormone treatments. The comprehensive results showed IDH, pentatricopeptide repeat-containing protein (PPRC), Unigene.16454, and caffeoyl-CoA O methyltransferase 5-like (CCOAOMT) are the top-ranked stable genes across all the samples. The stability of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the least during all experiments. Furthermore, the reliability of recommended reference gene was validated by the detection of porphobilinogen deaminase (HEMC) expression levels in chimeric leaves. Overall, this study provides appropriate reference genes under three specific experimental conditions and will be useful for future research on spatial and temporal regulation of gene expression and multiple hormone regulation pathways in Ananas comosus var. bracteatus.
An Integrative Analysis of Transcriptome and GWAS Data to Identify Potential Candidate Genes Influencing Meat Quality Traits in Pigs
Authors: Liu Xianxian , Zhang Junjie , Xiong Xinwei , Chen Congying , Xing Yuyun , Duan Yanyu , Xiao Shijun , Yang Bin , Ma Junwu
Published: October 2021

Understanding the genetic factors behind meat quality traits is of great significance to animal breeding and production. We previously conducted a genome-wide association study (GWAS) for meat quality traits in a White Duroc × Erhualian F2 pig population using Illumina porcine 60K SNP data. Here, we further investigate the functional candidate genes and their network modules associated with meat quality traits by integrating transcriptomics and GWAS information. Quantitative trait transcript (QTT) analysis, gene expression QTL (eQTL) mapping, and weighted gene co-expression network analysis (WGCNA) were performed using the digital gene expression (DGE) data from 493 F2 pig’s muscle and liver samples. Among the quantified 20,108 liver and 23,728 muscle transcripts, 535 liver and 1,014 muscle QTTs corresponding to 416 and 721 genes, respectively, were found to be significantly (p < 5 × 10−4) correlated with 22 meat quality traits measured on longissiums dorsi muscle (LM) or semimembranosus muscle (SM). Transcripts associated with muscle glycolytic potential (GP) and pH values were enriched for genes involved in metabolic process. There were 42 QTTs (for 32 genes) shared by liver and muscle tissues, of which 10 QTTs represent GP- and/or pH-related genes, such as JUNB, ATF3, and PPP1R3B. Furthermore, a genome-wide eQTL mapping revealed a total of 3,054 eQTLs for all annotated transcripts in muscle (p < 2.08 × 10−5), including 1,283 cis-eQTLs and 1771 trans-eQTLs. In addition, WGCNA identified five modules relevant to glycogen metabolism pathway and highlighted the connections between variations in meat quality traits and genes involved in energy process. Integrative analysis of GWAS loci, eQTL, and QTT demonstrated GALNT15/GALNTL2 and HTATIP2 as strong candidate genes for drip loss and pH drop from postmortem 45 min to 24 h, respectively. Our findings provide valuable insights into the genetic basis of meat quality traits and greatly expand the number of candidate genes that may be valuable for future functional analysis and genetic improvement of meat quality.
Kynurenic Acid and Its Analogue SZR-72 Ameliorate the Severity of Experimental Acute Necrotizing Pancreatitis
Authors: Balla Zsolt , Kormányos Eszter Sára , Kui Balázs , Bálint Emese Réka , Fűr Gabriella , Orján Erik Márk , Iványi Béla , Vécsei László , Fülöp Ferenc , Varga Gabriella , Harazin András , Tubak Vilmos , Deli Mária A. , Papp Csaba , Gácser Attila , Madácsy Tamara , Venglovecz Viktória , Maléth József , Hegyi Péter , Kiss Lóránd , Rakonczay Zoltán
Published: October 2021

The pathophysiology of acute pancreatitis (AP) is not well understood, and the disease does not have specific therapy. Tryptophan metabolite L-kynurenic acid (KYNA) and its synthetic analogue SZR-72 are antagonists of the N-methyl-D-aspartate receptor (NMDAR) and have immune modulatory roles in several inflammatory diseases. Our aims were to investigate the effects of KYNA and SZR-72 on experimental AP and to reveal their possible mode of action. AP was induced by intraperitoneal (i.p.) injection of L-ornithine-HCl (LO) in SPRD rats. Animals were pretreated with 75-300 mg/kg KYNA or SZR-72. Control animals were injected with physiological saline instead of LO, KYNA and/or SZR-72. Laboratory and histological parameters, as well as pancreatic and systemic circulation were measured to evaluate AP severity. Pancreatic heat shock protein-72 and IL-1β were measured by western blot and ELISA, respectively. Pancreatic expression of NMDAR1 was investigated by RT-PCR and immunohistochemistry. Viability of isolated pancreatic acinar cells in response to LO, KYNA, SZR-72 and/or NMDA administration was assessed by propidium-iodide assay. The effects of LO and/or SZR-72 on neutrophil granulocyte function was also studied. Almost all investigated laboratory and histological parameters of AP were significantly reduced by administration of 300 mg/kg KYNA or SZR-72, whereas the 150 mg/kg or 75 mg/kg doses were less or not effective, respectively. The decreased pancreatic microcirculation was also improved in the AP groups treated with 300 mg/kg KYNA or SZR-72. Interestingly, pancreatic heat shock protein-72 expression was significantly increased by administration of SZR-72, KYNA and/or LO. mRNA and protein expression of NMDAR1 was detected in pancreatic tissue. LO treatment caused acinar cell toxicity which was reversed by 250 µM KYNA or SZR-72. Treatment of acini with NMDA (25, 250, 2000 µM) did not influence the effects of KYNA or SZR-72. Moreover, SZR-72 reduced LO-induced H2O2 production of neutrophil granulocytes. KYNA and SZR-72 have dose-dependent protective effects on LO-induced AP or acinar toxicity which seem to be independent of pancreatic NMDA receptors. Furthermore, SZR-72 treatment suppressed AP-induced activation of neutrophil granulocytes. This study suggests that administration of KYNA and its derivative could be beneficial in AP.
Intranuclear Delivery of HIF-1α-TMD Alleviates EAE via Functional Conversion of TH17 Cells
Authors: Shin Jin-Su , Kim Ilkoo , Moon Jae-Seung , Ho Chun-Chang , Choi Min-Sun , Ghosh Sankar , Lee Sang-Kyou
Published: October 2021

T helper 17 (TH17) cells are involved in several autoimmune diseases such as multiple sclerosis (MS) and rheumatoid arthritis (RA). In addition to retinoic acid receptor-related orphan nuclear receptor gamma t (ROR-γt), hypoxia-inducible factor-1α (HIF-1α) is essential for the differentiation and inflammatory function of TH17 cells. To investigate the roles of HIF-1α in the functional regulation of TH17 cells under the normal physiological condition without genetic modification, the nucleus-transducible form of transcription modulation domain (TMD) of HIF-1α (ntHIF-1α-TMD) was generated by conjugating HIF-1α-TMD to Hph-1 protein transduction domain (PTD). ntHIF-1α-TMD was effectively delivered into the nucleus of T cells without cellular cytotoxicity. ntHIF-1α-TMD significantly blocked the differentiation of naïve T cells into TH17 cells in a dose-dependent manner via IL-17A and ROR-γt expression inhibition. However, T-cell activation events such as induction of CD69, CD25, and IL-2 and the differentiation potential of naïve T cells into TH1, TH2, or Treg cells were not affected by ntHIF-1α-TMD. Interestingly, TH17 cells differentiated from naïve T cells in the presence of ntHIF-1α-TMD showed a substantial level of suppressive activity toward the activated T cells, and the increase of Foxp3 and IL-10 expression was detected in these TH17 cells. When mRNA expression pattern was compared between TH17 cells and ntHIF-1α-TMD-treated TH17 cells, the expression of the genes involved in the differentiation and functions of TH17 cells was downregulated, and that of the genes necessary for immune-suppressive functions of Treg cells was upregulated. When the mice with experimental autoimmune encephalomyelitis (EAE) were treated with ntHIF-1α-TMD with anti-IL-17A mAb as a positive control, the therapeutic efficacy of ntHIF-1α-TMD in vivo was comparable with that of anti-IL-17A mAb, and ntHIF-1α-TMD-mediated therapeutic effect was contributed by the functional conversion of TH17 cells into immune-suppressive T cells. The results in this study demonstrate that ntHIF-1α-TMD can be a new therapeutic reagent for the treatment of various autoimmune diseases in which TH17 cells are dominant and pathogenic T cells.
Pathology and Immunity After SARS-CoV-2 Infection in Male Ferrets Is Affected by Age and Inoculation Route
Authors: van de Ven Koen , van Dijken Harry , Wijsman Lisa , Gomersbach Angéla , Schouten Tanja , Kool Jolanda , Lenz Stefanie , Roholl Paul , Meijer Adam , van Kasteren Puck B. , de Jonge Jørgen
Published: October 2021

Improving COVID-19 intervention strategies partly relies on animal models to study SARS-CoV-2 disease and immunity. In our pursuit to establish a model for severe COVID-19, we inoculated young and adult male ferrets intranasally or intratracheally with SARS-CoV-2. Intranasal inoculation established an infection in all ferrets, with viral dissemination into the brain and gut. Upon intratracheal inoculation only adult ferrets became infected. However, neither inoculation route induced observable COVID-19 symptoms. Despite this, a persistent inflammation in the nasal turbinates was prominent in especially young ferrets and follicular hyperplasia in the bronchi developed 21 days post infection. These effects -if sustained- might resemble long-COVID. Respiratory and systemic cellular responses and antibody responses were induced only in animals with an established infection. We conclude that intranasally-infected ferrets resemble asymptomatic COVID-19 and possibly aspects of long-COVID. Combined with the increasing portfolio to measure adaptive immunity, ferrets are a relevant model for SARS-CoV-2 vaccine research.
The ASC Speck and NLRP3 Inflammasome Function Are Spatially and Temporally Distinct
Authors: Nagar Abhinit , Rahman Tabassum , Harton Jonathan A.
Published: October 2021

Although considered the ternary inflammasome structure, whether the singular, perinuclear NLRP3:ASC speck is synonymous with the NLRP3 inflammasome is unclear. Herein, we report that the NLRP3:ASC speck is not required for nigericin-induced inflammasome activation but facilitates and maximizes IL-1β processing. Furthermore, the NLRP3 agonists H2O2 and MSU elicited IL-1β maturation without inducing specks. Notably, caspase-1 activity is spatially distinct from the speck, occurring at multiple cytoplasmic sites. Additionally, caspase-1 activity negatively regulates speck frequency and speck size, while speck numbers and IL-1β processing are negatively correlated, cyclical and can be uncoupled by NLRP3 mutations or inhibiting microtubule polymerization. Finally, when specks are present, caspase-1 is likely activated after leaving the speck structure. Thus, the speck is not the NLRP3 inflammasome itself, but is instead a dynamic structure which may amplify the NLRP3 response to weak stimuli by facilitating the formation and release of small NLRP3:ASC complexes which in turn activate caspase-1.
Diel to Seasonal Variation of Picoplankton in the Tropical South China Sea
Authors: Chen Tzong-Yueh , Lai Chao-Chen , Tai Jen-Hua , Ko Chia-Ying , Shiah Fuh-Kwo
Published: October 2021

Eight diel surveys on picoplankton (Prochlorococcus, Synechococcus, picoeukaryotes, and heterotrophic bacteria) abundance at the South East Asian Time-Series Station (SEATS; 18°N; 116°E) were conducted during the period of 2010 to 2014. The results indicated that Prochlorococcus and picoeukaryotes showed a subsurface maximum in warm seasons (spring, summer, and fall) and were abundant at the surface in the cold season (winter). Synechococcus and heterotrophic bacteria exhibited higher cell numbers at the surface and decreased with depth throughout the year. Although not all, some clear diel patterns for picoplankton were observed. Picophytoplankton usually peaked in the nighttime; picoeukaryotes peaked at ~7 to 8 p.m., followed by Synechococcus (peaking at 1 a.m.) and Prochlorococcus (peaking at 2 a.m.). Unlike these picoautotrophs, heterotrophic bacteria could peak either at dusk (i.e., 7 p.m.) or at noon. Seasonally, Prochlorococcus was more abundant in the warm than the cold seasons, while Synechococcus and picoeukaryotes showed blooms in the winter of 2013 and 2011, respectively. Heterotrophic bacteria showed no significant seasonality. Regression analysis indicated that ~73% of the diel-to-seasonal variation of the euphotic zone depth-integrated picophytoplankton biomass (i.e., PicoBeu) could be explained by the changes of the mixed-layer depth (MLD), and this suggested that inorganic nutrient supply could be the major controlling factor in their growth. The strong linear relationship (coefficient of determination, R2 of 0.83, p < 0.01) between sea surface temperature (SST) and PicoBeu implied, for the first time, a potential of using satellite-based SST to trace the biomass of picophytoplankton in the pelagic areas of the northern South China Sea.
Salt Marsh and Tidal Flat Area Distributions Along Three Estuaries
Authors: Kleinhans Maarten G. , Douma Harke , Addink Elisabeth A. , Coumou Laura , Deggeller Tessa , Jentink Robert , Paree Edwin , Cleveringa Jelmer
Published: October 2021

Estuarine landscapes form through interactions between fluvio-coastal processes and ecological processes within the boundaries imposed by hard substrate layers and man-made dikes and dams. As estuaries are ecologically valuable areas, monitoring and quantification of trends in habitats is needed for objective comparison and management. However, datasets of tidal flat and saltmarsh habitats along entire estuaries are scarce. The objective was to compare trends of biogeomorphological areas and habitat transitions along three estuaries in the Netherlands and assess whether these are generally comparable or mainly determined by system-specific histories. We present data for these estuaries obtained by automated classification of false-color aerial imagery. The automated method allows objective mapping of entire estuaries at unprecedented resolution. The estuaries are dominated by subtidal areas and tidal flats. The tidal flats have similar area along the estuaries while saltmarsh area decreases. Collective lengths of ecologically important transitions between saltmarsh, low-energy tidal flats and water differ more between the estuaries. These variations are due to presence of mid-channel bars and shore-connected embayments. Saltmarsh area is mainly determined by the different formation and embankment histories of the estuaries, embayments and side-branches. Much of the past saltmarsh flanking the estuaries was lost due to past land reclamation. In one system, ecologically important low-energy tidal flats are reduced by a sudden decrease of tidal amplitude, causing increase of subtidal area at the cost of intertidal area. Large areas of high-energy tidal flats in one estuary remain unexplained. The automated method can be applied in other estuaries, provided that high-quality areal imagery is available. Extensions of the data to other estuaries would allow for system-scale trend comparison between estuaries of ecologically relevant biogeomorphological characteristics.
Efficacy of Conbercept in the Treatment of Choroidal Neovascularization Secondary to Pathologic Myopia
Authors: Lu Hui , Yue Tao , Liu Na , Wang Zuo-Fen , Zhai Gai-Xia , Mi Dong-Ming , Zhang Jing , Wang Shao-Peng
Published: October 2021

Purpose: To observe the clinical efficacy of conbercept in the treatment of choroidal neovascularization (CNV) secondary to pathologic myopia.

Methods: We used retrospective analysis of the clinical data of 20 patients (24 eyes) with pathologic myopia choroidal neovascularization (PM-CNV). All patients were treated with intravitreal injection of conbercept 0.5 mg (0.05 ml), a vascular endothelial growth factor (VEGF) receptor fusion protein, and all patients completed at least 6 months of follow-up. Fundus, best corrected visual acuity (BCVA), fundus fluorescein angiography (FFA), optical coherence tomography (OCT), multifocal electroretinogram (mfERG) were assessed before and after treatment. Primary outcome was the functional change in amplitude by mfERG and secondary outcome was the structural change in central macular thickness (CRT) by OCT. The CNV area, leakage of CNV lesions, ocular and systemic adverse events were observed before and after treatment.

Results: The BCVA were 64.33 ± 10.83 letters, 65.42 ± 11.24 letters, 67.67 ± 7.07 letters after treatment 1, 3, 6 month, respectively, which showed improvement compared with the baseline (P < 0.05). The CRT decreased significantly from 308.50 ± 45.48 μm to 219.63 ± 30.27 μm, 221.33 ± 40.65 μm, 220.96 ± 33.09 μm after treatment 1, 3, 6 month, respectively (P < 0.05). The P1 response of mfERG amplitude improved from 40.71 ± 9.69 nv/deg2 to 50.67 ± 9.48 nv/deg2, 54.92 ± 8.45 nv/deg2, 55.67 ± 6.74 nv/deg2 after treatment 1, 3, 6 month, respectively (P < 0.05). After 6 months of treatment, the leakage of CNV lesions disappeared in 20 (83.3%) eyes, and the leakage area of CNV lesions was significantly reduced in 4 (16.7%) eyes.

Conclusion: The intravitreal injection of conbercept significantly reduced CRT and the CNV area, inhibited the leakage of CNV, improved the BCVA, increased the response of mfERG amplitude, and restored the retinal function. The intravitreal injection of conbercept can change the morphology and function of the macular in PM-CNV, which is safe and effective for the treatment of PM-CNV.
Genetic Polymorphisms and Clinical Features in Diabetic Patients With Fatty Liver: Results From a Single-Center Experience in Southern Italy
Authors: Villani Rosanna , Magnati Grazia Pia , De Girolamo Giuseppe , Sangineto Moris , Romano Antonino Davide , Cassano Tommaso , Serviddio Gaetano
Published: October 2021

Genetic background may be involved in the promotion and progression of non-alcoholic fatty liver disease (NAFLD). Previous studies have suggested that the single nucleotide polymorphisms (SNPs) may be associated with the specific clinical features in the patients with hepatic steatosis; however, data on the patients with diabetes from Southern Italy are lacking. We enrolled 454 patients and 260 of them had type 2 diabetes. We studied the PNPLA3 rs738409, LPIN1 rs13412852, KLF6 rs3750861, SOD2 rs4880, TM6SF2 rs58542926, and ZNF624 rs12603226 SNPs and their distribution in the study population. Lipid profile, liver stiffness, and kidney function were also studied to understand the potential role of the SNPs in the development of clinical phenotypes. No differences were observed in the distribution of polymorphisms between the diabetic and non-diabetic subjects. Carriers of risk allele G for PNPLA3 rs738409 SNP showed a lower mean value of serum triglycerides and a higher liver stiffness. Risk allele for KLF6 rs3750861 and SOD2 rs4880 polymorphism had a lower estimated glomerular filtration rate (eGFR) value, whereas no differences in the glucose and glycated hemoglobin level were observed in the subgroups by the different genotypes. Genetic polymorphisms are useful to identify the patients at higher risk of development of liver fibrosis and lower eGFR values in the patients with diabetes and NAFLD. Their use in clinical practice may help the clinicians to identify the patients who require a more strict follow-up program.
Investigating Fungi-Derived Bioactive Molecules as Inhibitor of the SARS Coronavirus Papain Like Protease: Computational Based Study
Authors: Belachew Aweke Mulu , Feyisa Asheber , Mohamed Seid Belay , W/Mariam Jerusalem Fekadu
Published: October 2021

Due to the rapid growth of the COVID-19 pandemic and its outcomes, developing a remedy to fight the predicament is critical. So far, it has infected more than 214,468,601 million people and caused the death of 4,470,969 million people according to the August 27, 2021, World Health Organization's (WHO) report. Several studies have been published on both computational and wet-lab approaches to develop antivirals for COVID-19, although there has been no success yet. However, the wet-lab approach is laborious, expensive, and time-consuming, and computational techniques have screened the activity of bioactive compounds from different sources with less effort and cost. For this investigation, we screened the binding affinity of fungi-derived bioactive molecules toward the SARS coronavirus papain-like protease (PLpro) by using computational approaches. Studies showed that protease inhibitors can be very effective in controlling virus-induced infections. Additionally, fungi represent a vast source of bioactive molecules, which could be potentially used for antiviral therapy. Fifty fungi-derived bioactive compounds were investigated concerning SARS-CoV-2 PLpro by using Auto Dock 4.2.1, Gromacs 2018. 2, ADMET, Swiss-ADME, FAF-Drugs 4.023, pKCSM, and UCLA-DOE server. From the list of the screened bioactive compounds, Dihydroaltersolanol C, Anthraquinone, Nigbeauvin A, and Catechin were selected with the Auto-Dock results of −8.68, −7.52, −10.46, and −10.58 Kcal/mol, respectively, based on their binding affinity compared to the reference drug. We presented the drug likeliness, toxicity, carcinogenicity, and mutagenicity of all compounds using ADMET analysis. They interacted with the amino acid residues, Gly163, Trp106, Ser111, Asp164, and Cys270, through hydrogen bonds. The root-mean-square deviation (RMSD), root-mean-square fluctuations (RMSF), solvent-accessible surface area (SASA), and radius of gyration (Rg) values revealed a stable interaction. From the overall analyses, we can conclude that Dihydroaltersolanol C, Anthraquinone, Nigbeauvin A, and Catechin are classified as promising candidates for PLpro, thus potentially useful in developing a medicine for COVID-19.
Age-Related Hearing Loss With Tinnitus and Physical Frailty Influence the Overall and Domain-Specific Quality of Life of Chinese Community-Dwelling Older Adults
Authors: Zhang Weibin , Ruan Jian , Zhang Ruxin , Zhang Min , Hu Xiuhua , Yu Zhuowei , Han Zhao , Ruan Qingwei
Published: October 2021

Objective: To investigate the impact of the severity of age-related hearing loss (ARHL) and tinnitus, presence of ARHL and/or tinnitus, and physical frailty on the health-related quality of life (HRQoL) and domain-specific HRQoL in Chinese community-dwelling older adults.

Design: This was a cross-sectional study of a community-based cohort.

Participants: We evaluated Chinese older adults (n = 429, 183 men and 246 women) aged ≥ 58years.

Measurements: The severity of HL and tinnitus were measured using pure-tone audiometry and the Tinnitus Handicap Inventory (THI), respectively. Physical frailty was measured using the five-item Fried scale. HRQoL was assessed using the Assessment of Quality of Life-8-Dimension (AQoL-8D) multi-attribute utility instrument (35 HRQoL items and eight domain-specific HRQoL subcategories). Low HRQoL (HRQoL score or subscores in the highest quintile) was used as a dependent variable in logistic regression analyses adjusted for demographic (Model 1) and health-related (Model 2) and psychosocial (Model 3) confounders.

Results: Age-related hearing loss severity was an independent determinant of senses in the physical dimension of HRQoL after adjusting for all covariates. Tinnitus severity was significantly associated with HRQoL and with independent living, senses, and pain in the physical dimension after adjusting for demographic and health-related covariates and was still associated with independent living and senses after adjusting for all covariates. The presence of ARHL and/or tinnitus was significantly associated with independent living and senses in the physical dimension after adjusting for all the covariates. Physical frailty was an independent determinant of HRQoL, independent living, and pain in the physical dimension and with mental health, happiness, and coping in the psychosocial dimension after adjusting for demographic and health-related covariates. The association with HRQoL, independent living, and pain in the physical dimension, and with happiness and coping in the psychosocial dimension remained significant after adjusting for the covariates. Depressive symptoms, social dysfunction, and a number of comorbidities were critical determinants of psychosocial HRQoL.

Conclusion: Physical frailty has a stronger and more profound effect on HRQoL, particularly on independent living and pain in the physical dimension and on happiness and coping in the psychosocial dimension. Domain-specific HRQoL should be considered in the management of patients with ARHL with tinnitus and physical frailty.

Clinical Trial Registration:www.ClinicalTrials.gov, identifier: NCT2017K020.
Flavonoid Library Screening Reveals Kaempferol as a Potential Antiviral Agent Against African Swine Fever Virus
Authors: Arabyan Erik , Hakobyan Astghik , Hakobyan Tamara , Grigoryan Rafaella , Izmailyan Roza , Avetisyan Aida , Karalyan Zaven , Jackman Joshua A. , Ferreira Fernando , Elrod Charles C. , Zakaryan Hovakim
Published: October 2021

Naturally occurring plant flavonoids are a promising class of antiviral agents to inhibit African swine fever virus (ASFV), which causes highly fatal disease in pigs and is a major threat to the swine industry. Currently known flavonoids with anti-ASFV activity demonstrate a wide range of antiviral mechanisms, which motivates exploration of new antiviral candidates within this class. The objective of this study was to determine whether other flavonoids may significantly inhibit ASFV infection in vitro. We performed a cell-based library screen of 90 flavonoids. Our screening method allowed us to track the development of virus-induced cytopathic effect by MTT in the presence of tested flavonoids. This screening method was shown to be robust for hit identification, with an average Z-factor of 0.683. We identified nine compounds that inhibit ASFV Ba71V strain in Vero cells. Among them, kaempferol was the most potent and exhibited dose-dependent inhibition, which occurred through a virostatic effect. Time-of-addition studies revealed that kaempferol acts on the entry and post-entry stages of the ASFV replication cycle and impairs viral protein and DNA synthesis. It was further identified that kaempferol induces autophagy in ASFV-infected Vero cells, which is related to its antiviral activity and could be partially abrogated by the addition of an autophagy inhibitor. Kaempferol also exhibited dose-dependent inhibition of a highly virulent ASFV Arm/07 isolate in porcine macrophages. Together, these findings support that kaempferol is a promising anti-ASFV agent and has a distinct antiviral mechanism compared to other anti-ASFV flavonoids.
Effects of Nitrogen and Phosphorus Limitation on Fatty Acid Contents in Aspergillus oryzae
Authors: Lv Gongbo , Xu Ying , Tu Yayi , Cheng Xiaojie , Zeng Bin , Huang Jianhua , He Bin
Published: October 2021

Aspergillus oryzae, commonly known as koji mold, has been widely used for the large-scale production of food products (sake, makgeolli, and soy sauce) and can accumulate a high level of lipids. In the present study, we showed the dynamic changes in A. oryzae mycelium growth and conidia formation under nitrogen and phosphorus nutrient stress. The fatty acid profile of A. oryzae was determined and the content of unsaturated fatty acid was found increased under nitrogen and phosphorus limitation. Oleic acid (C18:1), linoleic acid (C18:2), and γ-linolenic acid (C18:3) production were increased on five nitrogen and phosphorus limitation media, especially on nitrogen deep limitation and phosphorus limitation group, showing a 1. 2–, 1. 6–, and 2.4-fold increment, respectively, compared with the control. Transcriptomic analysis showed the expression profile of genes related to nitrogen metabolism, citrate cycle, and linoleic acid synthesis, resulting in the accumulation of unsaturated fatty acid. qRT-PCR results further confirmed the reliability and availability of the differentially expressed genes obtained from the transcriptome analysis. Our study provides a global transcriptome characterization of the nitrogen and phosphorus nutrient stress adaptation process in A. oryzae. It also revealed that the molecular mechanisms of A. oryzae respond to nitrogen and phosphorus stress. Our finding facilitates the construction of industrial strains with a nutrient-limited tolerance.
Modulating Drought Stress Response of Maize by a Synthetic Bacterial Community
Authors: Armanhi Jaderson Silveira Leite , de Souza Rafael Soares Correa , Biazotti Bárbara Bort , Yassitepe Juliana Erika de Carvalho Teixeira , Arruda Paulo
Published: October 2021

Plant perception and responses to environmental stresses are known to encompass a complex set of mechanisms in which the microbiome is involved. Knowledge about plant physiological responses is therefore critical for understanding the contribution of the microbiome to plant resilience. However, as plant growth is a dynamic process, a major hurdle is to find appropriate tools to effectively measure temporal variations of different plant physiological parameters. Here, we used a non-invasive real-time phenotyping platform in a one-to-one (plant–sensors) set up to investigate the impact of a synthetic community (SynCom) harboring plant-beneficial bacteria on the physiology and response of three commercial maize hybrids to drought stress (DS). SynCom inoculation significantly reduced yield loss and modulated vital physiological traits. SynCom-inoculated plants displayed lower leaf temperature, reduced turgor loss under severe DS and a faster recovery upon rehydration, likely as a result of sap flow modulation and better water usage. Microbiome profiling revealed that SynCom bacterial members were able to robustly colonize mature plants and recruit soil/seed-borne beneficial microbes. The high-resolution temporal data allowed us to record instant plant responses to daily environmental fluctuations, thus revealing the impact of the microbiome in modulating maize physiology, resilience to drought, and crop productivity.
Rapid Construction of an Infectious Clone of the Zika Virus, Strain ZKC2
Authors: Qin Zhiran , Chen Yangyang , Yu Jianhai , He Xiaoen , Liu Xuling , Zhu Li , Wu Qinghua , Wan Chengsong , Zhang Bao , Zhao Wei
Published: October 2021

Zika virus (ZIKV) has had detrimental effects on global public health in recent years. This is because the management of the disease has been limited, in part because its pathogenic mechanisms are not yet completely understood. Infectious clones are an important tool that utilize reverse genetics; these can be used to modify the ZIKV genomic RNA at the DNA level. A homologous recombination clone was used to construct pWSK29, a low copy plasmid that contained sequences for a T7 promoter, the whole genome of ZIKV ZKC2 strain, and a hepatitis delta virus ribozyme. High fidelity PCR was then used to amplify the T7 transcription template. The transcript was then transfected into susceptible cells via lipofection to recover the ZIKV ZKC2 strain. Finally, the virulence of rZKC2 was evaluated both in vitro and in vivo. The rZKC2 was successfully obtained and it showed the same virulence as its parent, the ZIKV ZKC2 strain (pZKC2), both in vitro and in vivo. The 3730 (NS2A-D62G) mutation site was identified as being important, since it had significant impacts on rZKC2 recovery. The 4015 (NS2A, A157V) mutation may reduce virus production by increasing the interferon type I response. In this study, one of the earliest strains of ZIKV that was imported into China was used for infectious clone construction and one possible site for antiviral medication development was discovered. The use of homologous recombination clones, of PCR products as templates for T7 transcription, and of lipofection for large RNA transfection could increase the efficiency of infectious clone construction. Our infectious clone provides an effective tool which can be used to explore the life cycle and medical treatment of ZIKV.
One Health Approaches to Trace Mycobacterium leprae’s Zoonotic Potential Through Time
Authors: Urban Christian , Blom Alette A. , Pfrengle Saskia , Walker-Meikle Kathleen , Stone Anne C. , Inskip Sarah A. , Schuenemann Verena J.
Published: October 2021

Hansen’s disease (leprosy), mainly caused by infection withMycobacterium leprae, has accompanied humanity for thousands of years. Although currently rare in Europe, there are over 200,000 new infections annually in South East Asia, Africa, and South America. Over the years many disciplines – palaeopathology, ancient DNA and other ancient biomolecules, and history – have contributed to a better understanding of leprosy’s past, in particular its history in medieval Europe. We discuss their contributions and potential, especially in relation to the role of inter-species transmission, an unexplored phenomenon in the disease’s history. Here, we explore the potential of interdisciplinary approaches that understand disease as a biosocial phenomenon, which is a product of both infection withM. lepraeand social behaviours that facilitate transmission and spread. Genetic evidence ofM. lepraeisolated from archaeological remains combined with systematic zooarchaeological and historical analysis would not only identify when and in what direction transmission occurred, but also key social behaviours and motivations that brought species together. In our opinion, this combination is crucial to understand the disease’s zoonotic past and current potential.
Improvement of the Biosynthesis of Resveratrol in Endophytic Fungus (Alternaria sp. MG1) by the Synergistic Effect of UV Light and Oligomeric Proanthocyanidins
Authors: Lu Yao , Shi Junling , Zhao Xixi , Song Yuyang , Qin Yi , Liu Yanlin
Published: October 2021

Resveratrol, a natural polyphenol compound with multiple bioactivities, is widely used in food and pharmaceutical industry. Endophytic fungus Alternaria sp. MG1, as a native producer of resveratrol, shows increasing potential application. However, strategies for improvement of the biosynthesis of resveratrol in this species are still scarce. In this study, different elicitors were used to investigate their effect on the biosynthesis of resveratrol in MG1 and the induction mechanism. Ultrasound and sodium butyrate had no effect and slight inhibition on the resveratrol production and related gene expression, respectively. UV radiation and co-culture with Phomopsis sp. XP-8 significantly promoted the biosynthesis of resveratrol with the highest production (240.57μg/l) coming from UV 20min. Co-culture altered the profiles of secondary metabolites in MG1 by promoting and inhibiting the synthesis of stilbene and lignin compounds, respectively, and generating new flavonoids ((+/−)-taxifolin, naringin, and (+)-catechin). Oligomeric proanthocyanidins (OPC) also showed an obviously positive influence, leading to an increase in resveratrol production by 10 to 60%. Two calcium-dependent protein kinases (CDPK) were identified, of which CDPK1 was found to be an important regulatory factor of OPC induction. Synergistic treatment of UV 20min and 100μm OPC increased the production of resveratrol by 70.37% compared to control and finally reached 276.31μg/l.
Predictive Value of Preoperative Profiling of Serum Metabolites for Emergence Agitation After General Anesthesia in Adult Patients
Authors: Wang Qian , Zhou Jiansuo , Liu Taotao , Yang Ning , Mi Xinning , Han Dengyang , Han Yongzheng , Chen Lei , Liu Kaixi , Zheng Hongcai , Zhang Jing , Lin Xiaona , Li Yitong , Hong Jingshu , Li Zhengqian , Guo Xiangyang
Published: October 2021

Background:Emergence agitation (EA) in adult patients under general anesthesia leads to increased postoperative complications and heavy medical burden. Unfortunately, its pathogenesis has not been clarified until now. The purpose of the present study was to explore the relationship between preoperative serum metabolites and EA.

Methods:We used an untargeted metabolic analysis method to investigate the different metabolomes in the serum of EA patients and non-EA patients undergoing elective surgical procedures after the induction of general anesthesia. A Richmond Agitation–Sedation Scale score ≥ +2 was diagnosed as EA during postoperative emergence. Non-EA patients were matched with EA patients according to general characteristics. Preoperative serum samples of the two groups were collected to investigate the association between serum metabolites and EA development.

Results:The serum samples of 16 EA patients with 34 matched non-EA patients were obtained for metabolic analysis. After screening and alignment with databases, 31 altered metabolites were detected between the two groups. These metabolites were mainly involved in the metabolism of lipids, purines, and amino acids. Analyses of receiver-operating characteristic curves showed that the preoperative alterations of choline, cytidine, glycerophosphocholine, L-phenylalanine, oleamide, and inosine may be associated with adult EA.

Conclusion:Multiple metabolic abnormalities (including those for lipids, purines, and amino acids) and other pathological processes (e.g., neurotransmitter imbalance and oxidative stress) may contribute to EA. Several altered metabolites in serum before surgery may have predictive value for EA diagnosis. This study might afford new metabolic clues for the understanding of EA pathogenesis.