bluuebunny/crossref_metadata_2025 huggingface dataset:
Studi Kualitatif Strategi Koping Penderita HIV AIDS di Kota Bandung
Authors: Salami Salami , Muvira Annisa Ayu , Yualita Perla
Published: March 2021

HIV/AIDS merupakan penyakit kronis yang dapat menimbulkan dampak negatif berupa masalah fisik, psikis, sosial, dan spiritual sehingga mengakibatkan orang dengan HIV/AIDS hidup dengan penuh tekanan karena isu negatif dan stigmatisasi. Hal ini dapat menjadi masalah bagi kejiwaan penderitanya sehingga memerlukan mekanisme koping ke arah adaptive. Strategi koping menentukan keberhasilan individu dalam menghadapi tekanan. Penelitian ini bertujuan mengeksplorasi strategi koping pada penderita HIV AIDS dengan menggunakan metode penelitian deskriptif kualitatif. Pengumpulan data menggunakan teknik in-depth interview terhadap 6 partisipan secara purposive sampling. Teknik analisis data yang digunakan adalah analysis interactive Miles dan Huberman. Hasil penelitian didapatkan 6 tema yaitu Pasrah pada Tuhan; Tidak ingin memikirkan penyakit; pengalihan masalah; Berupaya untuk bangkit; Upaya mengatasi masalah; dan Menyembunyikan status kesehatan. Berdasarkan penelitian ini dapat disimpulkan bahwa keenam partisipan memiliki kecenderungan melakukan emotion-focused coping. Bagi perawat diharapkan dapat menjalankan strategi intervensi dengan memberikan konseling maupun edukasi strategi koping yang adaptif kepada penderita dan keluarganya.
Identification of Stemness-Related Genes for Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma by Integrated Bioinformatics Analysis
Authors: Guo Hongjun , Wang Siqiao , Ju Min , Yan Penghui , Sun Wenhuizi , Li Zhenyu , Wu Siyu , Lin Ruoyi , Xian Shuyuan , Yang Daoke , Wang Jun , Huang Zongqiang
Published: March 2021

Background

Invasion and metastasis of cervical cancer are the main factors affecting the prognosis of patients with cervical squamous cell carcinoma (CESC). Therefore, it is of vital importance to find novel biomarkers that are associated with CESC invasion and metastasis, which will aid in the amelioration of individualized therapeutic methods for advanced patients.

Methods

The gene expression profiles of 10 metastatic and 116 non-metastatic samples were downloaded from The Cancer Genome Atlas (TCGA), where differentially expressed genes (DEGs) were defined. Weighted gene correlation network analysis (WGCNA) was employed to identify the stemness-related genes (SRGs). Univariate and multivariate regression analyses were used to identify the most significant prognostic key genes. Differential expression analysis of transcription factor (TF) and Gene Set Variation Analysis (GSVA) were utilized to explore the potential upstream regulation of TFs and downstream signaling pathways, respectively. Co-expression analysis was performed among significantly enriched TFs, key SRGs, and signaling pathways to construct a metastasis-specific regulation network in CESC. Connectivity Map (CMap) analysis was performed to identify bioactive small molecules which might be potential inhibitors for the network. Additionally, direct regulatory patterns of key genes were validated by ChIP-seq and ATAC-seq data.

Results

DEGs in yellow module acquired via WGCNA were defined as key genes which were most significantly related to mRNAsi. A multivariate Cox regression model was constructed and then utilized to explore the prognostic value of key SRGs by risk score. Area under curve (AUC) of the receiver operating characteristic (ROC) curve was 0.842. There was an obvious co expression pattern between the TF NR5A2 and the key gene VIM (R = 0.843, p < 0.001), while VIM was also significantly co-expressed with hallmark epithelial mesenchymal transition (EMT) signaling pathway (R = 0.318, p < 0.001). Naringenin was selected as the potential bioactive small molecule inhibitor for metastatic CESC based on CMap analysis.

Conclusions

VIM positively regulated by NR5A2 affected EMT signaling pathways in metastatic CESC, and naringenin was the inhibitor for the treatment of metastatic CESC via suppressing cancer stemness. This hypothetical signaling axis and potential inhibitors provide biomarkers and novel therapeutic targets for metastatic CESC.
Regulation of the SIAH2-HIF-1 Axis by Protein Kinases and Its Implication in Cancer Therapy
Authors: Xu Dazhong , Li Cen
Published: March 2021

The cellular response to hypoxia is a key biological process that facilitates adaptation of cells to oxygen deprivation (hypoxia). This process is critical for cancer cells to adapt to the hypoxic tumor microenvironment resulting from rapid tumor growth. Hypoxia-inducible factor 1 (HIF-1) is a transcription factor and a master regulator of the cellular response to hypoxia. The activity of HIF-1 is dictated primarily by its alpha subunit (HIF-1α), whose level and/or activity are largely regulated by an oxygen-dependent and ubiquitin/proteasome-mediated process. Prolyl hydroxylases (PHDs) and the E3 ubiquitin ligase Von Hippel-Lindau factor (VHL) catalyze hydroxylation and subsequent ubiquitin-dependent degradation of HIF-1α by the proteasome. Sevenin AbsentiaHomolog 2 (SIAH2), a RING finger-containing E3 ubiquitin ligase, stabilizes HIF-1α by targeting PHDs for ubiquitin-mediated degradation by the proteasome. This SIAH2-HIF-1 signaling axis is important for maintaining the level of HIF-1α under both normoxic and hypoxic conditions. A number of protein kinases have been shown to phosphorylate SIAH2, thereby regulating its stability, activity, or substrate binding. In this review, we will discuss the regulation of the SIAH2-HIF-1 axis via phosphorylation of SIAH2 by these kinases and the potential implication of this regulation in cancer biology and cancer therapy.
Chondroitinase ABC Promotes Axon Regeneration and Reduces Retrograde Apoptosis Signaling in Lamprey
Authors: Hu Jianli , Rodemer William , Zhang Guixin , Jin Li-Qing , Li Shuxin , Selzer Michael E.
Published: March 2021

Paralysis following spinal cord injury (SCI) is due to failure of axonal regeneration. It is believed that axon growth is inhibited by the presence of several types of inhibitory molecules in central nervous system (CNS), including the chondroitin sulfate proteoglycans (CSPGs). Many studies have shown that digestion of CSPGs with chondroitinase ABC (ChABC) can enhance axon growth and functional recovery after SCI. However, due to the complexity of the mammalian CNS, it is still unclear whether this involves true regeneration or only collateral sprouting by uninjured axons, whether it affects the expression of CSPG receptors such as protein tyrosine phosphatase sigma (PTPσ), and whether it influences retrograde neuronal apoptosis after SCI. In the present study, we assessed the roles of CSPGs in the regeneration of spinal-projecting axons from brainstem neurons, and in the process of retrograde neuronal apoptosis. Using the fluorochrome-labeled inhibitor of caspase activity (FLICA) method, apoptotic signaling was seen primarily in those large, individually identified reticulospinal (RS) neurons that are known to be “bad-regenerators.” Compared to uninjured controls, the number of all RS neurons showing polycaspase activity increased significantly at 2, 4, 8, and 11 weeks post-transection (post-TX). ChABC application to a fresh TX site reduced the number of polycaspase-positive RS neurons at 2 and 11 weeks post-TX, and also reduced the number of active caspase 3-positive RS neurons at 4 weeks post-TX, which confirmed the beneficial role of ChABC treatment in retrograde apoptotic signaling. ChABC treatment also greatly promoted axonal regeneration at 10 weeks post-TX. Correspondingly, PTPσ mRNA expression was reduced in the perikaryon. Previously, PTPσ mRNA expression was shown to correlate with neuronal apoptotic signaling at 2 and 10 weeks post-TX. In the present study, this correlation persisted after ChABC treatment, which suggests that PTPσ may be involved more generally in signaling axotomy-induced retrograde neuronal apoptosis. Moreover, ChABC treatment caused Akt activation (pAkt-308) to be greatly enhanced in brain post-TX, which was further confirmed in individually identified RS neurons. Thus, CSPG digestion not only enhances axon regeneration after SCI, but also inhibits retrograde RS neuronal apoptosis signaling, possibly by reducing PTPσ expression and enhancing Akt activation.
The Leishmania donovani LDBPK_220120.1 Gene Encodes for an Atypical Dual Specificity Lipid-Like Phosphatase Expressed in Promastigotes and Amastigotes; Substrate Specificity, Intracellular Localizations, and Putative Role(s)
Authors: Papadaki Amalia , Tziouvara Olympia , Kotopouli Anastasia , Koumarianou Petrina , Doukas Anargyros , Rios Pablo , Tardieux Isabelle , Köhn Maja , Boleti Haralabia
Published: March 2021

The intracellular protozoan parasites of the Leishmania genus are responsible for Leishmaniases, vector borne diseases with a wide range of clinical manifestations. Leishmania (L.) donovani causes visceral leishmaniasis (kala azar), the most severe of these diseases. Along their biological cycle, Leishmania parasites undergo distinct developmental transitions including metacyclogenesis and differentiation of metacyclic promastigotes (MPs) to amastigotes. Metacyclogenesis inside the phlebotomine sandfly host’s midgut converts the procyclic dividing promastigotes to non-dividing infective MPs eventually injected into the skin of mammalian hosts and phagocytosed by macrophages where the MPs are converted inside modified phagolysosomes to the intracellular amastigotes. These developmental transitions involve dramatic changes in cell size and shape and reformatting of the flagellum requiring thus membrane and cytoskeleton remodeling in which phosphoinositide (PI) signaling and metabolism must play central roles. This study reports on the LDBPK_220120.1 gene, the L. donovani ortholog of LmjF.22.0250 from L. major that encodes a phosphatase from the “Atypical Lipid Phosphatases” (ALPs) enzyme family. We confirmed the expression of the LDBPK_220120.1 gene product in both L. donovani promastigotes and axenic amastigotes and showed that it behaves in vitro as a Dual Specificity P-Tyr and monophosphorylated [PI(3)P and PI(4)P] PI phosphatase and therefore named it LdTyrPIP_22 (Leishmaniad onovani Tyrosine PI Phosphatase, gene locus at chromosome 22). By immunofluorescence confocal microscopy we localized the LdTyrPIP_22 in several intracellular sites in the cell body of L. donovani promastigotes and amastigotes and in the flagellum. A temperature and pH shift from 25°C to 37°C and from pH 7 to 5.5, induced a pronounced recruitment of LdTyrPIP_22 epitopes to the flagellar pocket and a redistribution around the nucleus. These results suggest possible role(s) for this P-Tyr/PI phosphatase in the regulation of processes initiated or upregulated by this temperature/pH shift that contribute to the developmental transition from MPs to amastigotes inside the mammalian host macrophages.
Explore the Protective Role of Obesity in the Progression of Myocardial Infarction
Authors: Zhao Siyuan , Cao Rongyuan , Zhang Shuhua , Kang Yan
Published: March 2021

Obesity has been shown as a risk factor to increase the incidence of myocardial infarction (MI). However, obesity has also been linked to the decreased mortality of acute MI with unknown mechanisms. Here, we firstly used large-scale literature data mining to identify obesity downstream targets and MI upstream regulators with polarity, based on which an obesity-MI regulatory network was constructed. Then, a gene set enrichment analysis was conducted to explore the functional profile of the genes involved in the obesity-MI regulatory networks. After that, a mega-analysis using MI RNA expression datasets was conducted to test the expression of obesity-specific genes in MI patients, followed by a shortest-path analysis to explore any potential gene-MI association. Our results suggested that obesity could inhibit 11 MI promoters, including NPPB, NPPA, IRS1, SMAD3, MIR155, ADRB1, AVP, MAPK14, MC3R, ROCK1, and COL3A1, which were mainly involved in blood pressure-related pathways. Our study suggested that obesity could influence MI progression by driving multiple genes associated with blood pressure regulation. Moreover, PTH could be a novel obesity driven gene associated with the pathogenesis of MI, which needs further validation.
Improving Acceptability and Uptake Behavior for Internet-Based Cognitive-Behavioral Therapy
Authors: Molloy Anthony , Ellis Donovan M. , Su Langting , Anderson Page L.
Published: March 2021

Internet-based cognitive behavioral therapy (iCBT) programs have the potential to improve access to mental healthcare, but they are not viewed as acceptable nor widely utilized by the general public. This study tested whether two acceptance-facilitating interventions improved acceptability and uptake-related behavior for therapist assisted and self-guided iCBT. Participants were randomly assigned to read a treatment rationale for iCBT (vs. a brief definition) and to receive a small financial incentive (or not) for seeking more information about evidence-based iCBT programs. Participants (N = 662) were a diverse group recruited from a University participant pool and the surrounding community. Participants completed standardized measures of attitudes toward and outcome expectancy for iCBT and a single question about willingness to use it and were given the opportunity to get information about accessing evidence-based iCBT programs. A series of MANCOVAs showed small, positive effects of the treatment rationale on attitudes and outcome expectancy for both self-guided and therapist-assisted iCBT, but not for willingness to use it. A hierarchical logistic regression model found no effect of the treatment rationale or financial incentive on whether participants sought additional information about how to access iCBT, although psychopathology symptoms and identifying as White or multiracial were positively associated with information-seeking. Inconsistent with past research, participants rated therapist-assisted and self-guided iCBT as equally acceptable. Participants recruited from the community reported greater willingness to use iCBT than University students. These results underscore the urgent need for further research toward improving the acceptability and uptake of iCBT so that it may better fulfill its potential to fill the gap in unmet mental health need.
Reconstructing Past Elevations From Triple Oxygen Isotopes of Lacustrine Chert: Application to the Eocene Nevadaplano, Elko Basin, Nevada, United States
Authors: Ibarra Daniel E. , Kukla Tyler , Methner Katharina A. , Mulch Andreas , Chamberlain C. Page
Published: March 2021

Triple oxygen isotope measurements are an emerging tool in paleoclimate reconstructions. In this contribution we develop the application of triple oxygen isotope measurements to lacustrine sediments to reconstruct past elevations. We focus on a well-constrained sample set from the Eocene North American Cordillera (Cherty Limestone Formation, Elko Basin, NV, United States, 42–43.5 Ma) on the east side of the elevated Nevadaplano. We present triple oxygen isotope measurements on freshwater lacustrine chert samples from the Cherty Limestone Formation. Across an evaporation trend spanning 6.5‰ in δ18O values we observe a negative correlation with Δ′17O ranging from −0.066 to −0.111‰ (λRL= 0.528), with an empirical slope (λchert, δ′17O vs. δ′18O) of 0.5236. Additionally, we present new carbonate clumped isotope (Δ47) temperature results on the overlying fluvial-lacustrine Elko Formation, which indicate an error-weighted mean temperature of 32.5 ± 3.8°C (1σ), and evaporatively enriched lake water spanning δ18O values of −3.7 to +3.5‰ (VSMOW). Paired chert and carbonate δ18O values demonstrate that co-equilbrium among the carbonate and chert phases is unlikely. Thus, as also previously suggested, it is most likely that Elko Basin chert formed during early diagenesis in equilbirium with pore waters that reflect evaporatively18O-enriched lake water. Using this scenario we apply a model for back-calculating unevaporated water composition to derive a source water of δ′18O = −16.1‰ (VSMOW), similar to modern local meteoric waters but lower than previous work on paired δ18O- δD measurements from the same chert samples. Further, this back-calculated unevaporated source water is higher than those derived using δD measurements of Late Eocene hydrated volcanic glass from the Elko Basin (average δ′18O equivalent of approximately −18.4‰, VSMOW). This suggests, assuming Eocene meteoric water Δ′17O values similar to today (∼0.032‰), either that: (1) the hypsometric mean elevation recorded by the lacustrine Cherty Limestone was lower than that derived from the average of the volcanic glass δD measurements alone; or (2) there was hydrogen exchange in volcanic glass with later low δD meteoric fluids. Nonetheless, our new findings support a relatively high (∼2.5–3 km) plateau recorded in the Elko Basin during the mid-Eocene.
Experimental Test of the Combined Effects of Water Availability and Flowering Time on Pollinator Visitation and Seed Set
Authors: Gallagher M. Kate , Campbell Diane R.
Published: March 2021

Climate change is likely to alter both flowering phenology and water availability for plants. Either of these changes alone can affect pollinator visitation and plant reproductive success. The relative impacts of phenology and water, and whether they interact in their impacts on plant reproductive success remain, however, largely unexplored. We manipulated flowering phenology and soil moisture in a factorial experiment with the subalpine perennialMertensia ciliata(Boraginaceae). We examined responses of floral traits, floral abundance, pollinator visitation, and composition of visits by bumblebees vs. other pollinators. To determine the net effects on plant reproductive success, we also measured seed production and seed mass. Reduced water led to shorter, narrower flowers that produced less nectar. Late flowering plants produced fewer and shorter flowers. Both flowering phenology and water availability influenced pollination and reproductive success. Differences in flowering phenology had greater effects on pollinator visitation than did changes in water availability, but the reverse was true for seed production and mass, which were enhanced by greater water availability. The probability of receiving a flower visit declined over the season, coinciding with a decline in floral abundance in the arrays. Among plants receiving visits, both the visitation rate and percent of non-bumblebee visitors declined after the first week and remained low until the final week. We detected interactions of phenology and water on pollinator visitor composition, in which plants subject to drought were the only group to experience a late-season resurgence in visits by solitary bees and flies. Despite that interaction, net reproductive success measured as seed production responded additively to the two manipulations of water and phenology. Commonly observed declines in flower size and reward due to drought or shifts in phenology may not necessarily result in reduced plant reproductive success, which inM. ciliataresponded more directly to water availability. The results highlight the need to go beyond studying single responses to climate changes, such as either phenology of a single species or how it experiences an abiotic factor, in order to understand how climate change may affect plant reproductive success.
Comparative Analyses of Full-Length Transcriptomes Reveal Gnetum luofuense Stem Developmental Dynamics
Authors: Hou Chen , Lian Huiming , Cai Yanling , Wang Yingli , Liang Dongcheng , He Boxiang
Published: March 2021

Genus Gnetum, of which the majority species are pantropical liana, have broad industrial uses including for string, nets, and paper production. Although numerous studies have investigated anatomical structures during stem development, the underlying molecular mechanisms that regulate this developmental trajectory in Gnetum species remain poorly understood. A total of 12 full-length transcriptomes were generated from four stem developmental stages of an arborescent representative of this genus, Gnetum luofuense, using Oxford Nanopore Technologies. The results of this analysis reveal a total of 24,151 alternative splicing (AS) and 134,391 alternative polyadenylation events. A remarkably dynamic pattern of AS events, especially in the case of intron retentions, was found across the four developmental stages while no dynamic pattern was found among transcript numbers with varied poly(A) sites. A total of 728 long non-coding RNAs were also detected; the number of cis-regulated target genes dramatically increased while no changes were found among trans-regulated target genes. In addition, a K-means clustering analysis of all full-length transcripts revealed that primary growth is associated with carbohydrate metabolism and fungi defense, while secondary growth is closely linked with photosynthesis, nitrogen transportation, and leaf ontogenesis. The use of weighted gene co-expression network analysis as well as differentially expressed transcripts reveals that bHLH, GRF, and MYB-related transcription factors are involved in primary growth, while AP2/ERF, MYB, NAC, PLAZ, and bZIP participate in G. luofuense stem secondary growth. The results of this study provide further evidence that Nanopore sequencing technology provides a cost-effective method for generating full-length transcriptome data as well as for investigating seed plant organ development.
Type 2 Diabetes and Cognitive Status in the Health and Retirement Study: A Mendelian Randomization Approach
Authors: Ware Erin B. , Morataya Cristina , Fu Mingzhou , Bakulski Kelly M.
Published: March 2021

Background

Type 2 diabetes mellitus (T2DM) and dementia are leading causes of mortality and disability in the US. T2DM has been associated with dementia; however, causality has not been clearly established. This study tested inferred causality between T2DM and dementia status using a Mendelian randomization approach.

Methods

Participants (50+ years) from the 2010 wave of the Health and Retirement Study of European or African genetic ancestry were included (n = 10,322). History of T2DM was self-reported. Cognitive status (dementia, cognitive impairment non-dementia, or normal cognition) was defined from clinically validated cognitive assessments. Cumulative genetic risk for T2DM was determined using a polygenic score calculated from a European ancestry T2DM genome-wide association study by Xue et al. (2018). All models were adjusted for age, sex, education, APOE-ε4 carrier status, and genetic principal components. Multivariable logistic regression was used to test the association between cumulative genetic risk for T2DM and cognitive status. To test inferred causality using Mendelian randomization, we used the inverse variance method.

Results

Among included participants, 20.9% had T2DM and 20.7% had dementia or cognitive impairment. Among European ancestry participants, T2DM was associated with 1.66 times odds of cognitive impairment non-dementia (95% confidence interval: 1.55–1.77) relative to normal cognition. A one standard deviation increase in cumulative genetic risk for T2DM was associated with 1.30 times higher odds of T2DM (95% confidence interval: 1.10–1.52). Cumulative genetic risk for T2DM was not associated with dementia status or cognitive-impaired non-dementia in either ancestry (P > 0.05); lack of association here is an important assumption of Mendelian randomization. Using Mendelian randomization, we did not observe evidence for an inferred causal association between T2DM and cognitive impairment (odds ratio: 1.04; 95% confidence interval: 0.90–1.21).

Discussion

Consistent with prior research, T2DM was associated with cognitive status. Prevention of T2DM and cognitive decline are both critical for public health, however, this study does not provide evidence that T2DM is causally related to impaired cognition. Additional studies in other ancestries, larger sample sizes, and longitudinal studies are needed to confirm these results.
Investigating the Underlying Mechanisms of Circular RNAs and Their Application in Clinical Research of Cervical Cancer
Authors: Liu Jian , Zhu He , Fu Li , Xu Tianmin
Published: March 2021

Circular RNAs (circRNAs) are non-coding RNA molecules, and these are differentially expressed in various diseases, including cancer, suggesting that circRNAs can regulate certain diseases. CircRNAs can act as miRNAs sponges, RNA-binding protein (RBP) sponges, and translation regulators, and they can become an important part of the regulation of gene expression. Furthermore, because of their biomedical features in body fluids, such as high abundance, conservation, and stability, circRNAs are seen as potential biomarkers for various cancers. Cervical cancer (CC) is one of the main causes of cancer-related death in women, and there have been a large number of studies that analyze circRNAs as a new object to be evaluated in CC. Therefore, this review, by understanding the role of circRNAs in CC, may create innovative strategies in the future clinical diagnosis, treatment, and prognosis of CC and promote the development of personalized and highly accurate cancer therapy.
Genetic Association of Interleukin 33/ST2 Polymorphisms With Behcet’s Uveitis
Authors: Pei Minghang , Liu Xinshu , Yang Peizeng , Zhao Chan , Gao Fei , Qu Yi , Liang Anyi , Xiao Junyan , Zhang Meifen
Published: March 2021

Interleukin (IL)33, a member of the IL1 superfamily, functions as a nuclear factor and mediates biological effects by interacting with the ST2 receptor. Recent studies have described IL33 as an emerging pro-inflammatory cytokine in the immune system, and IL33/ST2 gene polymorphisms have been implicated in the pathogenesis of various immune diseases. However, the underlying mechanisms of IL33/ST2 in Behcet’s disease (BD) remain to be defined. Here, we investigated the association between IL33/ST2 gene polymorphisms and BD in 585 BD uveitis (BDU) patients and 834 healthy controls using Agena MassARRAY iPLEX platform. We found that rs3821204 was associated with the development of BDU. Moreover, the frequency of rs2210463 G allele was lower in patients with genital involvement. Association analysis revealed a much greater genetic difference between complete-type and incomplete-type BD groups, including three SNPs (rs7044343, rs1048274, and rs2210463). Our findings suggest that IL33/ST2 gene polymorphisms are involved in the pathogenesis of BDU. Different genetic backgrounds may exist in complete-type and incomplete-type BD patients.
Tree Shrew as a New Animal Model for the Study of Dengue Virus
Authors: Jiang Liming , Lu Caixia , Sun Qiangming
Published: March 2021

Dengue virus is a significant public health threat worldwide; however, the pathogenesis of dengue disease remains poorly understood due to lack of appropriate small animal models. Tree shrews are an emerging experimental animal model for the study of human diseases due to their resemblance of genetic characteristics to primate animals. Herein we report that dengue infection in tree shrews elicits resemble clinical symptoms as in humans. Dengue fever (△2°C> normal body temperature) developed in ~22% healthy Chinese tree shrews from 2 through 33 days after infection with a low dose (1 ∗ 104 PFU/animal) of dengue virus serotype 2 or 3 intravenously or subcutaneously. The dengue genomic RNA and neutralizing antibodies were detected in ~78% of animals at days 7 and 15 post infection respectively. The serum levels of liver enzymes including aspartate transaminase, alanine aminotransferase and alkaline phosphatase were elevated with peaks at day 7 after infection. Modest thrombocytopenia and a slight decrease in the white blood cell count were observed. Intriguingly, although viral RNA was barely detectable in the liver by 48 days after infection, it was still evident in the brain. The intra-brain bleeding lesions in the intravenous infection group were more severe than those in the subcutaneous infection group. Our data demonstrate that primary dengue virus infection in tree shrews causes resemble clinical disease as in humans and thus tree shrews may be a suitable model for the study of dengue disease pathogenesis.
Leaky Gut Driven by Dysbiosis Augments Activation and Accumulation of Liver Macrophages via RIP3 Signaling Pathway in Autoimmune Hepatitis
Authors: Zhang Hongxia , Liu Man , Zhong Weilong , Zheng Yanping , Li Yanni , Guo Liping , Zhang Yujie , Ran Ying , Zhao Jingwen , Zhou Lu , Wang Bangmao
Published: March 2021

The gut–liver axis has been increasingly recognized as a major autoimmunity modulator. However, the implications of intestinal barrier in the pathogenesis of autoimmune hepatitis (AIH) remain elusive. Here, we investigated the functional role of gut barrier and intestinal microbiota for hepatic innate immune response in AIH patients and murine models. In this study, we found that AIH patients displayed increased intestinal permeability and pronounced RIP3 activation of liver macrophages. In mice models, intestinal barrier dysfunction increased intestinal bacterial translocation, thus amplifying the hepatic RIP3-mediated innate immune response. Furthermore, GSK872 dampened RIP3 activation and ameliorated the activation and accumulation of liver macrophages in vitro and in vivo experiments. Strikingly, broad-spectrum antibiotic ablation significantly alleviated RIP3 activation and liver injury, highlighting the causal role of intestinal microbiota for disease progression. Our results provided a potentially novel mechanism of immune tolerance breakage in the liver via the gut-liver axis. In addition, we also explored the therapeutic and research potentials of regulating the intestinal microbiota for the therapy of AIH.
Proteomic Characterization, Biodistribution, and Functional Studies of Immune-Therapeutic Exosomes: Implications for Inflammatory Lung Diseases
Authors: Elashiry Mahmoud , Elsayed Ranya , Elashiry Mohamed M. , Rashid Mohammad H. , Ara Roxan , Arbab Ali S. , Elawady Ahmed R. , Hamrick Mark , Liu Yutao , Zhi Wenbo , Lucas Rudolf , Vazquez Jose , Cutler Christopher W.
Published: March 2021

Dendritic cell (DC)-derived exosomes (DC EXO), natural nanoparticles of endosomal origin, are under intense scrutiny in clinical trials for various inflammatory diseases. DC EXO are eobiotic, meaning they are well-tolerated by the host; moreover, they can be custom-tailored for immune-regulatory or -stimulatory functions, thus presenting attractive opportunities for immune therapy. Previously we documented the efficacy of immunoregulatory DCs EXO (regDCs EXO) as immunotherapy for inflammatory bone disease, in an in-vivo model. We showed a key role for encapsulated TGFβ1 in promoting a bone sparing immune response. However, the on- and off-target effects of these therapeutic regDC EXO and how target signaling in acceptor cells is activated is unclear. In the present report, therapeutic regDC EXO were analyzed by high throughput proteomics, with non-therapeutic EXO from immature DCs and mature DCs as controls, to identify shared and distinct proteins and potential off-target proteins, as corroborated by immunoblot. The predominant expression in regDC EXO of immunoregulatory proteins as well as proteins involved in trafficking from the circulation to peripheral tissues, cell surface binding, and transmigration, prompted us to investigate how these DC EXO are biodistributed to major organs after intravenous injection. Live animal imaging showed preferential accumulation of regDCs EXO in the lungs, followed by spleen and liver tissue. In addition, TGFβ1 in regDCs EXO sustained downstream signaling in acceptor DCs. Blocking experiments suggested that sustaining TGFβ1 signaling require initial interaction of regDCs EXO with TGFβ1R followed by internalization of regDCs EXO with TGFβ1-TGFβ1R complex. Finally, these regDCs EXO that contain immunoregulatory cargo and showed biodistribution to lungs could downregulate the main severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) target receptor, ACE2 on recipient lung parenchymal cells via TGFβ1 in-vitro. In conclusion, these results in mice may have important immunotherapeutic implications for lung inflammatory disorders.
Markers of T Cell Exhaustion and Senescence and Their Relationship to Plasma TGF-β Levels in Treated HIV+ Immune Non-responders
Authors: Shive Carey L. , Freeman Michael L. , Younes Souheil-Antoine , Kowal Corinne M. , Canaday David H. , Rodriguez Benigno , Lederman Michael M. , Anthony Donald D.
Published: March 2021

Background: Immune non-responders (INR) are HIV+, ART-controlled (>2 yrs) people who fail to reconstitute their CD4 T cell numbers. Systemic inflammation and markers of T cell senescence and exhaustion are observed in INR. This study aims to investigate T cell senescence and exhaustion and their possible association with soluble immune mediators and to understand the immune profile of HIV-infected INR. Selected participants were <50 years old to control for the confounder of older age.

Methods: Plasma levels of IL-6, IP10, sCD14, sCD163, and TGF-β and markers of T cell exhaustion (PD-1, TIGIT) and senescence (CD57, KLRG-1) were measured in ART-treated, HIV+ participants grouped by CD4 T cell counts (n = 63). Immune parameters were also measured in HIV-uninfected, age distribution-matched controls (HC; n = 30). Associations between T cell markers of exhaustion and senescence and plasma levels of immune mediators were examined by Spearman rank order statistics.

Results: Proportions of CD4 T cell subsets expressing markers of exhaustion (PD-1, TIGIT) and senescence (CD57, KLRG-1) were elevated in HIV+ participants. When comparing proportions between INR and IR, INR had higher proportions of CD4 memory PD-1+, EM CD57+, TEM TIGIT+ and CD8 EM and TEM TIGIT+ cells. Plasma levels of IL-6, IP10, and sCD14 were elevated during HIV infection. IP10 was higher in INR. Plasma TGF-β levels and CD4 cycling proportions of T regulatory cells were lower in INR. Proportions of CD4 T cells expressing TIGIT, PD-1, and CD57 positively correlated with plasma levels of IL-6. Plasma levels of TGF-β negatively correlated with proportions of TIGIT+ and PD-1+ T cell subsets.

Conclusions: INR have lower levels of TGF-β and decreased proportions of cycling CD4 T regulatory cells and may have difficulty controlling inflammation. IP10 is elevated in INR and is linked to higher proportions of T cell exhaustion and senescence seen in INR.
Transcriptional and Immunologic Correlates of Response to Pandemic Influenza Vaccine in Aviremic, HIV-Infected Children
Authors: de Armas Lesley R. , George Varghese , Filali-Mouhim Abdelali , Steel Courtney , Parmigiani Anita , Cunningham Coleen K. , Weinberg Adriana , Trautmann Lydie , Sekaly Rafick-Pierre , Cameron Mark J. , Pahwa Savita
Published: March 2021

People living with HIV (PWH) often exhibit poor responses to influenza vaccination despite effective combination anti-retroviral (ART) mediated viral suppression. There exists a paucity of data in identifying immune correlates of influenza vaccine response in context of HIV infection that would be useful in improving its efficacy in PWH, especially in younger individuals. Transcriptomic data were obtained by microarray from whole blood isolated from aviremic pediatric and adolescent HIV-infected individuals (4-25 yrs) given two doses of Novartis/H1N1 09 vaccine during the pandemic H1N1 influenza outbreak. Supervised clustering and gene set enrichment identified contrasts between individuals exhibiting high and low antibody responses to vaccination. High responders exhibited hemagglutination inhibition antibody titers >1:40 post-first dose and 4-fold increase over baseline. Baseline molecular profiles indicated increased gene expression in metabolic stress pathways in low responders compared to high responders. Inflammation-related and interferon-inducible gene expression pathways were higher in low responders 3 wks post-vaccination. The broad age range and developmental stage of participants in this study prompted additional analysis by age group (e.g. <13yrs and ≥13yrs). This analysis revealed differential enrichment of gene pathways before and after vaccination in the two age groups. Notably, CXCR5, a homing marker expressed on T follicular helper (Tfh) cells, was enriched in high responders (>13yrs) following vaccination which was accompanied by peripheral Tfh expansion. Our results comprise a valuable resource of immune correlates of vaccine response to pandemic influenza in HIV infected children that may be used to identify favorable targets for improved vaccine design in different age groups.
Mitochondrial Reactive Oxygen Species Regulate Immune Responses of Macrophages to Aspergillus fumigatus
Authors: Hatinguais Remi , Pradhan Arnab , Brown Gordon D. , Brown Alistair J. P. , Warris Adilia , Shekhova Elena
Published: March 2021

Reactive Oxygen Species (ROS) are highly reactive molecules that can induce oxidative stress. For instance, the oxidative burst of immune cells is well known for its ability to inhibit the growth of invading pathogens. However, ROS also mediate redox signalling, which is important for the regulation of antimicrobial immunity. Here, we report a crucial role of mitochondrial ROS (mitoROS) in antifungal responses of macrophages. We show that mitoROS production rises in murine macrophages exposed to swollen conidia of the fungal pathogen Aspergillus fumigatus compared to untreated macrophages, or those treated with resting conidia. Furthermore, the exposure of macrophages to swollen conidia increases the activity of complex II of the respiratory chain and raises mitochondrial membrane potential. These alterations in mitochondria of infected macrophages suggest that mitoROS are produced via reverse electron transport (RET). Significantly, preventing mitoROS generation via RET by treatment with rotenone, or a suppressor of site IQ electron leak, S1QEL1.1, lowers the production of pro-inflammatory cytokines TNF-α and IL-1β in macrophages exposed to swollen conidia of A. fumigatus. Rotenone and S1QEL1.1 also reduces the fungicidal activity of macrophages against swollen conidia. Moreover, we have established that elevated recruitment of NADPH oxidase 2 (NOX2, also called gp91phox) to the phagosomal membrane occurs prior to the increase in mitoROS generation. Using macrophages from gp91phox-/- mice, we have further demonstrated that NOX2 is required to regulate cytokine secretion by RET-associated mitoROS in response to infection with swollen conidia. Taken together, these observations demonstrate the importance of RET-mediated mitoROS production in macrophages infected with A. fumigatus.
Flagellin From Pseudomonas Aeruginosa Stimulates ATB0,+ Transporter for Arginine and Neutral Amino Acids in Human Airway Epithelial Cells
Authors: Barilli Amelia , Visigalli Rossana , Ferrari Francesca , Borsani Giuseppe , Dall'Asta Valeria , Rotoli Bianca Maria
Published: March 2021

At present, the central role played by arginine in the modulation of the inflammatory cellular responses is well-recognized, and many pro-inflammatory stimuli are known to modulate the expression and activity of its transmembrane transporters. In this regard, we have addressed the effects of bacterial flagellin from Pseudomonas aeruginosa (FLA-PA) on the uptake of the amino acid in human epithelial respiratory cells. Among the arginine transporters, only ATB0,+, y+L, and y+ were operative in bronchial epithelial Calu-3 cells under control conditions; however, only the expression and activity of ATB0,+ were stimulated upon incubation with flagellin, whereas those of systems y+L and y+ were not stimulated. As a result, this induction, in turn, led to an increase in the intracellular content of arginine without making any change to its metabolic pathway. In addition, flagellin upregulated the amount of other amino acids substrates of ATB0,+, in particular, all the essential amino acids, such as valine, isoleucine, and leucine, along with the non-essential glutamine. At the molecular level, these effects were directly referable to the stimulation of a toll-like receptor-5 (TLR5) signaling pathway and to the induction of nuclear factor-κB (NF-κB) transcription factor. An induction of ATB0,+ expression has been observed also in EpiAirway™, a model of primary human normal tracheal-bronchial epithelial cells that mimics the in vitro pseudostratified columnar epithelium of the airways. In this tissue model, the incubation with flagellin is associated with the upregulation of messenger RNAs (mRNAs) for the chemokine IL-8 and for the cytokines IL-6 and interleukin-1β (IL-1β); as for the latter, a marked secretion in the extracellular medium was also observed due to the concomitant activation of caspase-1. The overall findings indicate that, in human respiratory epithelium, flagellin promotes cellular responses associating the increase of intracellular amino acids through ATB0,+ with the activation of the inflammasome. Given the role of the ATB0,+ transporter as a delivery system for bronchodilators in human airway epithelial cells, its induction under inflammatory conditions gains particular relevance in the field of respiratory pharmacology.
Clinical Utility of the Detection of the Loss of the Mismatched HLA in Relapsed Hematological Patients After Haploidentical Stem Cell Transplantation With High-Dose Cyclophosphamide
Authors: Muñiz Paula , Kwon Mi , Carbonell Diego , Chicano María , Bailén Rebeca , Oarbeascoa Gillen , Suárez-González Julia , Andrés-Zayas Cristina , Menárguez Javier , Dorado Nieves , Gómez-Centurión Ignacio , Anguita Javier , Díez-Martín José Luis , Martínez-Laperche Carolina , Buño Ismael
Published: March 2021

Haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) with high-dose cyclophosphamide (PTCy) has resulted in a low incidence of graft-vs.-host disease (GVHD), graft failure, and non-relapse mortality. However, post-transplantation relapse remains a common cause of treatment failure in high-risk patients. Unraveling the mechanisms of relapse is therefore crucial for designing effective relapse treatment strategies. One of these mechanisms is the loss of the mismatched HLA on the recipient's leukemic cells. To study the incidence and clinical relevance of this phenomenon, we analyzed 181 patients treated with Haplo-HSCT with PTCy (2007–2019), of which 37 relapsed patients after transplantation. According to the kit employed for HLA-loss analysis, among 22 relapsed patients, we identified HLA loss at relapse in 6 of the 22 patients (27%) studied. Based on the results obtained, the genomic loss of HLA was more common in females than males (66 vs. 33%) and HLA-loss relapses occurred later than classical relapses (345 vs. 166 days). Moreover, the patients with HLA-loss had a greater presence of active disease at the time of transplantation and had undergone a larger number of treatment lines than the group with classical relapses (66 vs. 43% and 66 vs. 18%, respectively). Four of these relapses were studied retrospectively, while two were studied prospectively, the results of which could be considered for patient management. Additionally, two relapsed patients analyzed retrospectively had myeloid neoplasms. One patient had not undergone any treatment, and three had undergone donor lymphocyte infusions (DLIs) and chemotherapy. All presented severe GVHD and disease progression. In contrast, the two patients studied prospectively had a lymphoid neoplasm and were not treated with DLIs. One of them was treated with chemotherapy but died from disease progression, and the other patient underwent a second Haplo-HSCT from a different donor and is still alive. We can conclude that the detection of HLA-loss at the onset of relapse after Haplo-HSCT with PTCy could help in clinical practice to select appropriate rescue treatment, thereby avoiding the use of DLIs or a second transplantation from the same donor.
TIGIT-Fc as a Potential Therapeutic Agent for Fetomaternal Tolerance
Authors: Fu Wenyan , Cai Renfei , Ma Zetong , Li Tian , Lei Changhai , Zhao Jian , Hu Shi
Published: March 2021

The perfect synchronization of maternal immune-endocrine mechanisms and those of the fetus is necessary for a successful pregnancy. In this report, decidual immune cells at the maternal-fetal interface were detected that expressed TIGIT (T cell immunoreceptor with Ig and ITIM domains), which is a co-inhibitory receptor that triggers immunological tolerance. We generated recombinant TIGIT-Fc fusion proteins by linking the extracellular domain of TIGIT and silent Fc fragments. The treatment with TIGIT-Fc of human decidual antigen presenting cells (APCs), the decidual dendritic cells (dDCs), and decidual macrophages (dMϕs) increased the production of interleukin 10 and induced the decidua APCs to powerfully polarize the decidual CD4+ T cells toward a classic TH2 phenotype. We further proposed that Notch signaling shows a pivotal effect on the transcriptional regulation in decidual immune cell subsets. Moreover, the administration of TIGIT-Fc to CBA/J pregnant mice at preimplantation induced CD4+ forkhead box P3+ (Foxp3+) regulatory T cells and tolerogenic dendritic cells and increased pregnancy rates in an abortion-prone animal model stress. The results suggested the therapeutic potential of the TIGIT-Fc fusion protein in reinstating immune tolerance in failing pregnancies.
A Novel Autoantibody Induced by Bacterial Biofilm Conserved Components Aggravates Lupus Nephritis
Authors: Fu Wenyan , Liu Yu , Liu Fangjie , Liu Chenghua , Li Jingjing , Niu Jiali , Han Peng , Xu Dan , Hou Jiaojiao , Ma Yuanfang , Feng Jiannan , Li Zhanguo , Mu Rong , Yang Guang
Published: March 2021

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with multiple autoantibody production and often affects the kidneys, known as lupus nephritis. However, the mechanism underlying lupus nephritis development is unclear. Biofilms that protect bacteria from stress are ubiquitous in almost every environment. Here, we identified that a conserved peptide (HU1) derived from DNABII proteins, one of major bacterial biofilm components, was specifically recognized by sera from about 47% patients with SLE. Moreover, the serum anti-HU1 levels showed a significant positive correlation with lupus nephritis occurrence. Presence of antibodies against HU1 in pristane-induced mice aggravated lupus nephritis, although these antibodies also attenuated bacterial biofilm formation. We further identified that antibodies against HU1 cross-recognized protein disulfide isomerase (P4HB) located on the renal cell surface and inhibited the activities of this enzyme. Our findings reveal a novel mechanism underlying the development of lupus nephritis triggered by bacterial biofilms.