bluuebunny/crossref_metadata_2025 huggingface dataset:
Weighted Gene Co-Expression Network Analysis (WGCNA) Reveals the Functions of Syndecan-1 to Regulate Immune Infiltration by Influenced T Cells in Glioma
Authors: Zhong Jiacheng , Shi Shuang , Peng Wen , Liu Bei , Yang Biao , Niu Wenyong , Zhang Biao , Qin Chuan , Zhong Dong , Cui Hongjuan , Zhang Zhengbao , Sun Xiaochuan
Published: May 2022

Our previous studies shown that syndecan-1 (SDC1) may be a novel class of biomarkers for the diagnosis and treatment of glioma, but its specific roles and the in-depth molecular mechanism remain elusive. Here, we used Estimation of STromal and Immune cells in Malignant Tumor tissues using Expression data (ESTIMATE) algorithms and single-sample Gene Set Enrichment Analysis (ssGSEA) algorithms to evaluate the immune score of tumor samples and quantify the relative infiltration of immune cells in the tumor microenvironment (TME), respectively, in different data sets obtained from the Chinese Glioma Genome Atlas and The Cancer Gene Atlas. Next, we calculate the correlation of the immune score and immune cells with SDC1, respectively. To identify the specific process regulated by SDC1, the differentially expressed genes (DEGs) analysis between the high and low expression of SDC1 of glioma samples were used to discover the hub genes through Weighted Gene Coexpression Network Analysis (WGCNA). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed cardinal biological processes and pathways involved in genes and tumor grade correlation and survival analysis verified its significance in glioma. The results show that SDC1 is associated with the immune infiltration of glioma in the TME, especially activated CD4+T cells and CD8+T cells. The three data sets filter 8,887 DEGs, the genes in the blue modules were selected as hub genes in WGCNA. GO and KEGG analysis found eight genes in the blue modules involved in antigen processing and presentation in T cells in glioma. Kaplan–Meier estimator and log-rank test statistic determined that the introduced genes are associated with poor prognosis in glioma. Protein–protein network interaction analysis showed that SDC1 may regulate antigen processing and presentation through CTSL or CD4 in glioma. Finally, this study provided insights and clues for the next research direction of SDC1 and identified the key pathways and genes that might participate in the immune escape of glioma. These results might provide a new insight on the study of immune infiltration of glioma in the future.
Exome Sequencing Data Analysis and a Case-Control Study in Mexican Population Reveals Lipid Trait Associations of New and Known Genetic Variants in Dyslipidemia-Associated Loci
Authors: Jurado-Camacho Pedro A. , Cid-Soto Miguel A. , Barajas-Olmos Francisco , García-Ortíz Humberto , Baca-Peynado Paulina , Martínez-Hernández Angélica , Centeno-Cruz Federico , Contreras-Cubas Cecilia , González-Villalpando María Elena , Saldaña-Álvarez Yolanda , Salas-Martinez Guadalupe , Mendoza-Caamal Elvia C. , González-Villalpando Clicerio , Córdova Emilio J. , Orozco Lorena
Published: May 2022

Background: Plasma lipid levels are a major risk factor for cardiovascular diseases. Although international efforts have identified a group of loci associated with the risk of dyslipidemia, Latin American populations have been underrepresented in these studies.

Objective: To know the genetic variation occurring in lipid-related loci in the Mexican population and its association with dyslipidemia.

Methods: We searched for single-nucleotide variants in 177 lipid candidate genes using previously published exome sequencing data from 2838 Mexican individuals belonging to three different cohorts. With the extracted variants, we performed a case-control study. Logistic regression and quantitative trait analyses were implemented in PLINK software. We used an LD pruning using a 50-kb sliding window size, a 5-kb window step size and a r2 threshold of 0.1.

Results: Among the 34251 biallelic variants identified in our sample population, 33% showed low frequency. For case-control study, we selected 2521 variants based on a minor allele frequency ≥1% in all datasets. We found 19 variants in 9 genes significantly associated with at least one lipid trait, with the most significant associations found in the APOA1/C3/A4/A5-ZPR1-BUD13 gene cluster on chromosome 11. Notably, all 11 variants associated with hypertriglyceridemia were within this cluster; whereas variants associated with hypercholesterolemia were located at chromosome 2 and 19, and for low high density lipoprotein cholesterol were in chromosomes 9, 11, and 19. No significant associated variants were found for low density lipoprotein. We found several novel variants associated with different lipemic traits: rs3825041 in BUD13 with hypertriglyceridemia, rs7252453 in CILP2 with decreased risk to hypercholesterolemia and rs11076176 in CETP with increased risk to low high density lipoprotein cholesterol.

Conclusions: We identified novel variants in lipid-regulation candidate genes in the Mexican population, an underrepresented population in genomic studies, demonstrating the necessity of more genomic studies on multi-ethnic populations to gain a deeper understanding of the genetic structure of the lipemic traits.
The Four Key Genes Participated in and Maintained Atrial Fibrillation Process via Reprogramming Lipid Metabolism in AF Patients
Authors: Fang Yijin , Wu Yue , Liu Liangming , Wang Huadong
Published: May 2022

Atrial fibrillation (AF) is always in high incidence in the population, which can lead to serious complications. The structural and electrical remodeling of atrial muscle induced by inflammatory reaction or oxidative stress was considered as the major mechanism of AF. The treatment effect is not ideal based on current mechanisms. Recent studies demonstrated that lipid metabolism disorder of atrial muscle played an important role in the occurrence of AF. What key genes are involved is unclear. The purpose of the present study was to explore the lipid metabolism mechanism of AF. With the GEO database and the genomics of AF patients, metabolic related pathways and the key genes were analyzed. At the same time, the rat model of cecal ligation and puncture (CLP) was used to confirm the results. GSE 31821 and GSE 41177 were used as data sources, and the merged differentially expressed genes (DEGs) analysis showed that a total of 272 DEGs were found. GO annotation, KEGG, and gene set enrichment analysis (GSEA) showed that the fatty acid metabolism and the lipid biosynthetic process were involved in AF. Cholesterol biosynthesis, arachidonic acid metabolism, and the lipid droplet pathway were obviously increased in AF. Further analysis showed that four key genes, including ITGB1, HSP90AA1, CCND1, and HSPA8 participated in pathogenesis of AF regulating lipid biosynthesis. In CLP rats, metabolic profiling in the heart showed that the pyrimidine metabolism, the biosynthesis of unsaturated fatty acid metabolism, arginine and proline metabolism, and the fatty acid biosynthesis were involved. The four key genes were confirmed increased in the heart of CLP rats (p < 0.05 or 0.01). The results suggest that the lipid metabolism disorder participates in the occurrence of AF. ITGB1, HSP90AA1, CCND1, and HSPA8 are the key genes involved in the regulation of lipid biosynthesis.
Biological and Clinicopathological Characteristics of OPN in Cervical Cancers
Authors: Qin Shuhang , Yi Li , Liang Yanchun , Chen Yili , Wang Wei , Liao Yuandong , Zhang Chunyu , Huang Hua , Huang Jiaming , Yao Shuzhong
Published: May 2022

Background: Cervical cancer (CC) is the most common gynecological malignancy. Recently, an increasing number of studies have indicated that osteopontin (OPN) is a promising diagnostic and prognostic biomarker for CC. However, the biological role and detailed mechanism of OPN in CC remain unclear.

Methods: The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets and a clinical sample microarray were used in our study. To identify the clinicopathological characteristics of OPN in CC, we compared the expression of OPN between normal and CC tissue samples and analyzed the correlations between OPN expression and multiple clinicopathological features. To identify biological processes involving OPN, OPN-associated genes were screened with Pearson correlation analysis and applied in hallmark gene set enrichment analysis (GSEA). Additionally, leukocyte infiltration was evaluated based on OPN expression. Finally, OPN-related signaling pathways were identified by GSEA.

Results: OPN expression was higher in CC samples than in normal tissue samples and positively correlated with age, FIGO stage, tumor size, lymphovascular invasion and an unfavorable prognosis. OPN-associated genes were mainly enriched in the immune response, and increased OPN expression was accompanied by increased M2 macrophage infiltration. Additionally, OPN was correlated with hypoxia, high glycolytic metabolism, apoptosis, angiogenesis, epithelial-mesenchymal transition and multiple signaling pathways (the p53 pathway, the PI3K/Akt pathway, IL6/STAT3 signaling, mTORC1 signaling and KRAS signaling).

Conclusion: Our study showed that OPN is involved in immunological activities and multiple tumor processes, identifying it as a potential therapeutic target and useful prognostic factor in CC patients.
Digital Cell Atlas of Mouse Uterus: From Regenerative Stage to Maturational Stage
Authors: Zhang Leyi , Long Wenying , Xu Wanwan , Chen Xiuying , Zhao Xiaofeng , Wu Bingbing
Published: May 2022

Endometrium undergoes repeated repair and regeneration during the menstrual cycle. Previous attempts using gene expression data to define the menstrual cycle failed to come to an agreement. Here we used single-cell RNA sequencing data of C57BL/6J mice uteri to construct a novel integrated cell atlas of mice uteri from the regenerative endometrium to the maturational endometrium at the single-cell level, providing a more accurate cytological-based elucidation for the changes that occurred in the endometrium during the estrus cycle. Based on the expression levels of proliferating cell nuclear antigen, differentially expressed genes, and gene ontology terms, we delineated in detail the transitions of epithelial cells, stromal cells, and immune cells that happened during the estrus cycle. The transcription factors that shaped the differentiation of the mononuclear phagocyte system had been proposed, being Mafb, Irf7, and Nr4a1. The amounts and functions of immune cells varied sharply in two stages, especially NK cells and macrophages. We also found putative uterus tissue-resident macrophages and identified potential endometrial mesenchymal stem cells (high expression of Cd34, Pdgfrb, Aldh1a2) in vivo. The cell atlas of mice uteri presented here would improve our understanding of the transitions that occurred in the endometrium from the regenerative endometrium to the maturational endometrium. With the assistance of a normal cell atlas as a reference, we may identify morphologically unaffected abnormalities in future clinical practice. Cautions would be needed when adopting our conclusions, for the limited number of mice that participated in this study may affect the strength of our conclusions.
Identification and Expression Analysis of Calcium-Dependent Protein Kinases Gene Family in Potato Under Drought Stress
Authors: Dekomah Simon Dontoro , Wang Yihao , Qin Tianyuan , Xu Derong , Sun Chao , Yao Panfeng , Liu Yuhui , Bi Zhenzhen , Bai Jiangping
Published: May 2022

Calcium-dependent protein kinases (CDPKs) are a class of serine/threonine protein kinases encoded by several gene families that play key roles in stress response and plant growth and development. In this study, the BLAST method was used to search for protein sequences of the potato Calcium-dependent protein kinase gene family. The chromosome location, phylogeny, gene structures, gene duplication, cis-acting elements, protein-protein interaction, and expression profiles were analyzed. Twenty-five CDPK genes in the potato genome were identified based on RNA-seq data and were clustered into four groups (I-IV) based on their structural features and phylogenetic analysis. The result showed the composition of the promoter region of the StCDPKs gene, including light-responsive elements such as Box4, hormone-responsive elements such as ABRE, and stress-responsive elements such as MBS. Four pairs of segmental duplications were found in StCDPKs genes and the Ka/Ks ratios were below 1, indicating a purifying selection of the genes. The protein-protein interaction network revealed defense-related proteins such as; respiratory burst oxidase homologs (RBOHs) interacting with potato CDPKs. Transcript abundance was measured via RT-PCR between the two cultivars and their relative expression of CDPK genes was analyzed after 15, 20, and 25 days of drought. There were varied expression patterns of StCDPK3/13/21 and 23, between the two potato cultivars under mannitol induced-drought conditions. Correlation analysis showed that StCDPK21/22 and StCDPK3 may be the major differentially expressed genes involved in the regulation of malondialdehyde (MDA) and proline content in response to drought stress, opening a new research direction for genetic improvement of drought resistance in potato.
Gpr174 Knockout Alleviates DSS-Induced Colitis via Regulating the Immune Function of Dendritic Cells
Authors: Wei Wei , Mu Sucheng , Han Yi , Chen Yao , Kuang Zhongshu , Wu Xingyue , Luo Yue , Tong Chaoyang , Zhang Yiqun , Yang Yilin , Song Zhenju
Published: May 2022

Background

Dysfunction of the immune system would disturb the intestinal homeostasis and lead to inflammatory bowel disease (IBD). Dendritic cells (DCs) help maintain intestinal homeostasis and immediately respond to pathogens or injuries once the mucosa barriers are destroyed during IBD. G protein-coupled receptors(GPR)174 is an essential regulator of immunity that is widely expressed in most immune cells, including DCs. However, the role of GPR174 in regulating the immune function of DC in colitis has not been investigated.

Methods

Dextran sodium sulfate (DSS) was administered to establish the mice colitis model. Data of weight, length of colon, disease activity index (DAI), and macroscopic scores were collected. The flow cytometry was used to detect the infiltrations of T cells and DCs, the mean fluorescence intensity (MFI) of CD80, CD86, CD40, and major histocompatibility complex-II (MHC-II). And T cells proliferataion was measured by carboxyfluorescein diacetate succinimidyl ester (CFSE). The expression of cytokines (tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-γ (IFN-γ), interleukin -4 (IL-4)) and GPR174 mRNA were measured by Elisa, quantitative polymerase chain reaction (qPCR), and immunofluorescence. RNA of bone-marrow-derived dendritic cells (BMDCs) was extracted for sequencing. Adoptive transfer of BMDCs was administrated intravenously.

Results

Gpr174-/- mice exposed to 3% DSS showed significant alleviation characterized by reduced loss of weight, more minor colon damage, and better DAI and macroscopic scores. The expression of pro-inflammatory cytokines (TNF-α, IL-6) decreased, while anti-inflammatory cytokine (IL-10) increased compared with WT mice. In vitro, Gpr174-/- BMDCs showed less maturity, with a declined expression of MHC-II, CD80, CD86 and reduced TNF-α, higher IL-10 after LPS stimulation. Gpr174-/- BMDCs were less capable of activating OT-II naïve CD4+ T cells than WT BMDCs and induced more Th0 cells to differentiate into Treg while less into Th1. Furthermore, the transcriptome sequencing analysis exhibited that Gpr174 participated in TNF-α (NF-κB) signaling, leukocyte transendothelial migration, and Th1/Th2 cell differentiation pathways. Adoptive transfer of Gpr174-/- BMDCs to WT mice ameliorated DSS-induced colitis.

Conclusion

Our study indicated that GPR174 was involved in the pathogenesis of IBD by regulating the maturation of the dendritic cells to maintain immune homeostasis. TNF-α (NF-κB) signaling pathway, leukocyte transendothelial migration, and Th1/Th2 cell differentiation pathways may be the target pathway.
Evaluation of Host Protein Biomarkers by ELISA From Whole Lysed Peripheral Blood for Development of Diagnostic Tests for Active Tuberculosis
Authors: Garlant Harriet N. , Ellappan Kalaiarasan , Hewitt Matthew , Perumal Prem , Pekeleke Simon , Wand Nadina , Southern Jo , Kumar Saka Vinod , Belgode Harish , Abubakar Ibrahim , Sinha Sanjeev , Vasan Seshadri , Joseph Noyal Mariya , Kempsell Karen E.
Published: May 2022

Tuberculosis (TB) remains a significant global health crisis and the number one cause of death for an infectious disease. The health consequences in high-burden countries are significant. Barriers to TB control and eradication are in part caused by difficulties in diagnosis. Improvements in diagnosis are required for organisations like the World Health Organisation (WHO) to meet their ambitious target of reducing the incidence of TB by 50% by the year 2025, which has become hard to reach due to the COVID-19 pandemic. Development of new tests for TB are key priorities of the WHO, as defined in their 2014 report for target product profiles (TPPs). Rapid triage and biomarker-based confirmatory tests would greatly enhance the diagnostic capability for identifying and diagnosing TB-infected individuals. Protein-based test methods e.g. lateral flow devices (LFDs) have a significant advantage over other technologies with regard to assay turnaround time (minutes as opposed to hours) field-ability, ease of use by relatively untrained staff and without the need for supporting laboratory infrastructure. Here we evaluate the diagnostic performance of nine biomarkers from our previously published biomarker qPCR validation study; CALCOCO2, CD274, CD52, GBP1, IFIT3, IFITM3, SAMD9L, SNX10 and TMEM49, as protein targets assayed by ELISA. This preliminary evaluation study was conducted to quantify the level of biomarker protein expression across latent, extra-pulmonary or pulmonary TB groups and negative controls, collected across the UK and India, in whole lysed blood samples (WLB). We also investigated associative correlations between the biomarkers and assessed their suitability for ongoing diagnostic test development, using receiver operating characteristic/area under the curve (ROC) analyses, singly and in panel combinations. The top performing single biomarkers for pulmonary TB versus controls were CALCOCO2, SAMD9L, GBP1, IFITM3, IFIT3 and SNX10. TMEM49 was also significantly differentially expressed but downregulated in TB groups. CD52 expression was not highly differentially expressed across most of the groups but may provide additional patient stratification information and some limited use for incipient latent TB infection. These show therefore great potential for diagnostic test development either in minimal configuration panels for rapid triage or more complex formulations to capture the diversity of disease presentations.
Human Melanoma-Associated Mast Cells Display a Distinct Transcriptional Signature Characterized by an Upregulation of the Complement Component 3 That Correlates With Poor Prognosis
Authors: Bahri Rajia , Kiss Orsolya , Prise Ian , Garcia-Rodriguez Karen M. , Atmoko Haris , Martínez-Gómez Julia M. , Levesque Mitchell P. , Dummer Reinhard , Smith Michael P. , Wellbrock Claudia , Bulfone-Paus Silvia
Published: May 2022

Cutaneous melanoma is one of the most aggressive human malignancies and shows increasing incidence. Mast cells (MCs), long-lived tissue-resident cells that are particularly abundant in human skin where they regulate both innate and adaptive immunity, are associated with melanoma stroma (MAMCs). Thus, MAMCs could impact melanoma development, progression, and metastasis by secreting proteases, pro-angiogenic factors, and both pro-inflammatory and immuno-inhibitory mediators. To interrogate the as-yet poorly characterized role of human MAMCs, we have purified MCs from melanoma skin biopsies and performed RNA-seq analysis. Here, we demonstrate that MAMCs display a unique transcriptome signature defined by the downregulation of the FcεRI signaling pathway, a distinct expression pattern of proteases and pro-angiogenic factors, and a profound upregulation of complement component C3. Furthermore, in melanoma tissue, we observe a significantly increased number of C3+ MCs in stage IV melanoma. Moreover, in patients, C3 expression significantly correlates with the MC-specific marker TPSAB1, and the high expression of both markers is linked with poorer melanoma survival. In vitro, we show that melanoma cell supernatants and tumor microenvironment (TME) mediators such as TGF-β, IL-33, and IL-1β induce some of the changes found in MAMCs and significantly modulate C3 expression and activity in MCs. Taken together, these data suggest that melanoma-secreted cytokines such as TGF-β and IL-1β contribute to the melanoma microenvironment by upregulating C3 expression in MAMCs, thus inducing an MC phenotype switch that negatively impacts melanoma prognosis.
Repositioning of the Angiotensin II Receptor Antagonist Candesartan as an Anti-Inflammatory Agent With NLRP3 Inflammasome Inhibitory Activity
Authors: Lin Wen-Yu , Li Lan-Hui , Hsiao Ya-Yun , Wong Wei-Ting , Chiu Hsiao-Wen , Hsu Hsien-Ta , Peng Yi-Jen , Ho Chen-Lung , Chernikov Oleg V. , Cheng Shu-Meng , Yang Shih-Ping , Hua Kuo-Feng
Published: May 2022

Aberrant activation of the NLRP3 inflammasome promotes the pathogenesis of many inflammatory diseases. The development of the NLRP3 inflammasome inhibitors from existing drugs for new therapeutic purposes is becoming more important. Candesartan is an angiotensin II receptor antagonist widely used as a blood pressure-lowering drug; however, the inhibitory potential of candesartan on the NLRP3 inflammasome has not yet been investigated. We demonstrated that candesartan significantly inhibited the NLRP3 inflammasome and pyroptosis in macrophages. Mechanistic analysis revealed that candesartan inhibited the expression of NLRP3 and proIL-1β by suppressing NF-κB activation and reducing the phosphorylation of ERK1/2 and JNK1/2. Candesartan reduced mitochondrial damage and inhibited the NLRP3 inflammasome assembly by suppressing NLRP3 binding to PKR, NEK7 and ASC. In addition, candesartan inhibited IL-1β secretion partially through autophagy induction. Furthermore, oral administration of candesartan reduced peritoneal neutrophil influx, NLRP3 and ASC expression in peritoneal cells, and lavage fluid concentrations of active caspase-1, IL-1β, IL-6 and MCP-1 in uric acid crystal-injected mice. These results indicated that candesartan has board anti-inflammatory effects and has the potential to be repositioned to ameliorate inflammatory diseases or NLRP3-associated complications.
The BLT Humanized Mouse Model as a Tool for Studying Human Gamma Delta T Cell-HIV Interactions In Vivo
Authors: Biradar Shivkumar , Agarwal Yash , Lotze Michael T. , Bility Moses T. , Mailliard Robbie B.
Published: May 2022

Gamma-delta (γδ) T cells recognize antigens in a major histocompatibility complex (MHC) independent and have cytotoxic capability. Human immunodeficiency virus (HIV) infection reduces the proportion of the Vδ2 cell subset compared to the Vδ1 cell subset of γδ T cells in the blood in most infected individuals, except for elite controllers. The capacity of Vδ2 T cells to kill HIV-infected targets has been demonstratedin vitro, albeitin vivoconfirmatory studies are lacking. Here, we provide the first characterization of γδ T cell-HIV interactions in bone marrow-liver-thymus (BLT) humanized mice and examined the immunotherapeutic potential of Vδ2 T cells in controlling HIV replicationin vivo. We demonstrate a reduced proportion of Vδ2 T cells and an increased proportion of Vδ1 T cells in HIV-infected BLT humanized mice, like in HIV-positive individuals. HIV infection in BLT humanized mice also impaired theex vivoexpansion of Vδ2 T cells, like in HIV-positive individuals. Adoptive transfer of activated Vδ2 T cells did not control HIV replication during cell-associated HIV transmission in BLT humanized mice but instead exacerbated viremia, suggesting that Vδ2 T cells may serve as early targets for HIV replication. Our findings demonstrate that BLT humanized mice can model γδ T cell-HIV interactionsin vivo.
Effectiveness and Durability of mRNA Vaccine-Induced SARS-CoV-2-Specific Humoral and Cellular Immunity in Severe Asthma Patients on Biological Therapy
Authors: Podrazil Michal , Taborska Pavla , Stakheev Dmitry , Rataj Michal , Lastovicka Jan , Vlachova Alena , Pohunek Petr , Bartunkova Jirina , Smrz Daniel
Published: May 2022

Coronavirus disease 2019 (COVID-19) vaccines effectively elicit humoral and cellular immunity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in healthy populations. This immunity decreases several months after vaccination. However, the efficacy of vaccine-induced immunity and its durability in patients with severe asthma on biological therapy are unknown. In this study, we evaluated the effectiveness and durability of mRNA vaccine-induced SARS-CoV-2-specific humoral and cellular immunity in severe asthma patients on biological therapy. The study included 34 patients with severe asthma treated with anti-IgE (omalizumab, n=17), anti-IL5 (mepolizumab, n=13; reslizumab, n=3), or anti-IL5R (benralizumab, n=1) biological therapy. All patients were vaccinated with two doses of the BNT162b2 mRNA vaccine with a 6-week interval between the doses. We found that this COVID-19 vaccination regimen elicited SARS-CoV-2-specific humoral and cellular immunity, which had significantly declined 6 months after receipt of the second dose of the vaccine. The type of biological treatment did not affect vaccine-elicited immunity. However, patient age negatively impacted the vaccine-induced humoral response. On the other hand, no such age-related impact on vaccine-elicited cellular immunity was observed. Our findings show that treatment of patients with severe asthma with biological therapy does not compromise the effectiveness or durability of COVID-19 vaccine-induced immunity.
Contemplating Dichotomous Nature of Gamma Delta T Cells for Immunotherapy
Authors: Bhat Jaydeep , Placek Katarzyna , Faissner Simon
Published: May 2022

γδ T cells are unconventional T cells, distinguished from αβ T cells in a number of functional properties. Being small in number compared to αβ T cells, γδ T cells have surprised us with their pleiotropic roles in various diseases. γδ T cells are ambiguous in nature as they can produce a number of cytokines depending on the (micro) environmental cues and engage different immune response mechanisms, mainly due to their epigenetic plasticity. Depending on the disease condition, γδ T cells contribute to beneficial or detrimental response. In this review, we thus discuss the dichotomous nature of γδ T cells in cancer, neuroimmunology and infectious diseases. We shed light on the importance of equal consideration for systems immunology and personalized approaches, as exemplified by changes in metabolic requirements. While providing the status of immunotherapy, we will assess the metabolic (and other) considerations for better outcome of γδ T cell-based treatments.
Role of COVID-19 Vaccines in SARS-CoV-2 Variants
Authors: Zhou Zhou , Zhu Yimiao , Chu Ming
Published: May 2022

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), is a threat to the health of the global population. As the result of a global effort in the determination of origin, structure, and pathogenesis of SARS-CoV-2 and its variants, particularly such the variant of concern as Delta Variant and Omicron Variant, the understanding of SARS-CoV-2 are deepening and the development of vaccines against SARS-CoV-2 are ongoing. Currently, AstraZeneca-Vaxzevria/SII-Covishield vaccine, Janssen-Ad26.COV2.S vaccine, Moderna-mRNA-1273 vaccine, Pfizer BioNTech-Comirnaty vaccine and Sinovac-CoronaVac vaccine have been listed as WHO Emergency Use Listing (EUL) Qualified Vaccines by WHO. Because of the antigen escape caused by the mutation in variants, the effectiveness of vaccines, which are currently the main means of prevention and treatment, has been affected by varying degrees. Herein, we review the current status of mutations of SARS-CoV-2 variants, the different approaches used in the development of COVID-19 vaccines, and COVID-19 vaccine effectiveness against SARS-CoV-2 variants.
A Versatile Hemolin With Pattern Recognitional Contributions to the Humoral Immune Responses of the Chinese Oak Silkworm Antheraea pernyi
Authors: He Xueshan , Zhou Tianyang , Cai Yuchen , Liu Yang , Zhao Siqi , Zhang Jinghai , Wang Xialu , Zhang Rong
Published: May 2022

Hemolin is a distinctive immunoglobulin superfamily member involved in invertebrate immune events. Although it is believed that hemolin regulates hemocyte phagocytosis and microbial agglutination in insects, little is known about its contribution to the humoral immune system. In the present study, we focused on hemolin in Antheraea pernyi (Ap-hemolin) by studying its pattern recognition property and humoral immune functions. Tissue distribution analysis demonstrated the mRNA level of Ap-hemolin was extremely immune-inducible in different tissues. The results of western blotting and biolayer interferometry showed recombinant Ap-hemolin bound to various microbes and pathogen-associated molecular patterns. In further immune functional studies, it was detected that knockdown of hemolin regulated the expression level of antimicrobial peptide genes and decreased prophenoloxidase activation in the A. pernyi hemolymph stimulated by microbial invaders. Together, these data suggest that hemolin is a multifunctional pattern recognition receptor that plays critical roles in the humoral immune responses of A. pernyi.
KPI5 Is Involved in the Regulation of the Expression of Antibacterial Peptide Genes and Hemolymph Melanization in the Silkworm, Bombyx mori
Authors: Heng Jingya , Liu Huawei , Xu Jiahui , Huang Xuan , Sun Xiaotong , Yang Runze , Xia Qingyou , Zhao Ping
Published: May 2022

Kunitz-type protease inhibitors (KPIs) are ubiquitously found in many organisms, and participate in various physiological processes. However, their function in insects remains to be elucidated. In the present study, we characterized and functionally analyzed silkworm KPI5. Sequence analysis showed that KPI5 contains 85 amino acids with six conserved cysteine residues, and the P1 site is a phenylalanine residue. Inhibitory activity and stability analyses indicated that recombinant KPI5 protein significantly inhibited the activity of chymotrypsin and was highly tolerant to temperature and pH. The spatio-temporal expression profile analysis showed that KPI5 was synthesized in the fat body and secreted into the hemolymph. In vivo induction analysis showed that the expression of KPI5 in the fat body was significantly upregulated by pathogen-associated molecular patterns (PAMPs). Binding assays suggested that KPI5 can bind to pathogens and PAMPs. In vitro pathogen growth inhibition assay and encapsulation analysis indicated that KPI5 can neither kill pathogenic bacteria directly nor promote the encapsulation of agarose beads by silkworm hemocytes. Recombinant protein injection test and CRISPR/Cas9-mediated knockdown showed that KPI5 promotes the expression of antimicrobial peptides (AMPs) in the fat body. Moreover, the survival rate of individuals in the KPI5 knockdown group was significantly lower than that of the control group after pathogen infection. Phenoloxidase (PO) activity assays showed that KPI5 significantly inhibited the hemolymph PO activity and melanization induced by PAMPs. These findings suggested that KPI5 plays a dual regulatory role in innate immunity by promoting the expression of antimicrobial peptides in the fat body and inhibiting hemolymph melanization. Our study furthers the understanding of the function of insect KPIs and provides new insights into the regulatory mechanism of insect immune homeostasis.
Research on Evaluation Index of High Temperature Performance of Rubberized Asphalt Binder
Authors: Xie Cheng , Luo Junhui , Zeng Lihao , Ren Tianzeng , Liu Haobin , Chen Jiangcai
Published: May 2022

The high-temperature rheological properties of rubberized asphalt and mixture were evaluated by frequency scanning, repeated creep recovery (RCR) test, temperature sweep test, Hamburg wheel tracking test (HWTT). Based on the Pearson correlation coefficient, the correlation of the mixture gradation and asphalt characteristics with the high-temperature stability of the mixture was analyzed. Finally, the gray correlation theory was applied to analyze the evaluation indexes of the high-temperature rheological properties of rubberized asphalt and mixture. The results show that the asphalt-stone ratio and the fractal dimension (Dc) of coarse aggregate have a significant correlation with the high-temperature performance of the mixture, and the mixture with a smaller asphalt-stone ratio and a higher percentage of coarse aggregate has a better high-temperature performance. The correlation degree of softening point, viscous stiffness modulus and permanent deformation with rubber-asphalt mixture is higher than 0.7, and are significantly higher than those of rotational viscosity at 180°C. Therefore, we recommend the use of permanent deformation, softening point, and viscous stiffness modulus to evaluate the high-temperature performance of rubberized asphalt mixture.
Evaluation of the Relationship Between Aniseikonia and Stereopsis Using a New Method
Authors: Xu Lingxian , Liu Lu , Wu Huang
Published: May 2022

Purpose

To investigate the influence of induced aniseikonia on stereopsis measured by contour-based and random-dot-based stereograms using a new method.

Methods

Unlike previous studies in which aniseikonia was induced using magnifiers, which potentially influenced the position of the test symbols in the half-view, here the image was magnified while maintaining each test symbol’s central position within the half-view. A phoropter and two 4K smartphones were used to measure stereopsis in seventeen young adults aged 20–28 years old. Stereopsis was tested using both contour-based and random-dot-based stereograms under overall or meridional aniseikonia with magnifications ranging from 2.5 to 30%. Repeated measures ANOVA was used to evaluate the effect of aniseikonia on stereopsis.

Results

Stereopsis decreased with an increase in aniseikonia magnification in the overall, horizontal, and vertical directions. Stereopsis values (log arcsec) increased from 1.29 ± 0.14 at baseline to 2.38 ± 0.16 with 30% overall aniseikonia of contour-based stereograms. In random-dot based stereograms, stereopsis values increased from 1.29 ± 0.16 at baseline to 2.24 ± 0.23 with 22.5% overall aniseikonia. Overall aniseikonia caused a significantly greater impairment on stereopsis as compared with the changes in meridional directions. In contour-based stereograms, vertical aniseikonia had significantly less impact on stereopsis than horizontal aniseikonia of identical magnification. The opposite phenomenon was found in random-dot-based stereograms.

Conclusion

Stereopsis decreased with an increase of magnification of induced aniseikonia. Magnifying patterns (overall, horizontal, or vertical) also significantly affected stereopsis. The conflicting impact of meridional aniseikonia on stereopsis measured by contour-based and random-dot-based stereograms may be associated with the uniqueness of the two test systems.
The Intrapopulation Genetic Diversity of RNA Virus May Influence the Sensitivity of Chlorine Disinfection
Authors: Kadoya Syun-suke , Urayama Syun-ichi , Nunoura Takuro , Hirai Miho , Takaki Yoshihiro , Kitajima Masaaki , Nakagomi Toyoko , Nakagomi Osamu , Okabe Satoshi , Nishimura Osamu , Sano Daisuke
Published: May 2022

RNA virus populations are not clonal; rather, they comprise a mutant swarm in which sequences are slightly different from the master sequence. Genetic diversity within a population (intrapopulation genetic diversity) is critical for RNA viruses to survive under environmental stresses. Disinfection has become an important practice in the control of pathogenic viruses; however, the impact of intrapopulation genetic diversity on the sensitivity of disinfection, defined as –log10 (postdisinfected infectious titer/predisinfected titer), has not been elucidated. In this study, we serially passaged populations of rhesus rotavirus. We demonstrated that populations with reduced chlorine sensitivity emerged at random and independently of chlorine exposure. Sequencing analysis revealed that compared with sensitive populations, less-sensitive ones had higher non-synonymous genetic diversity of the outer capsid protein gene, suggesting that changes in the amino acid sequences of the outer capsid protein were the main factors influencing chlorine sensitivity. No common mutations were found among less-sensitive populations, indicating that rather than specific mutations, the diversity of the outer capsid protein itself was associated with the disinfection sensitivity and that the disinfection sensitivity changed stochastically. Simulation results suggest that the disinfection sensitivity of a genetically diverse population is destabilized if cooperative viral clusters including multiple sequences are formed. These results advocate that any prevention measures leading to low intrapopulation genetic diversity are important to prevent the spread and evolution of pathogenic RNA viruses in society.
Metagenomic and Metabolomic Insights Into the Mechanism Underlying the Disparity in Milk Yield of Holstein Cows
Authors: Amin Abdulmumini B. , Zhang Lei , Zhang JiYou , Mao Shengyong
Published: May 2022

This study was conducted to investigate the metabolic mechanism underlying the disparity in the milk yield of Holstein cows. Eighteen lactating Holstein cows in their second parity and 56 (±14.81 SD) days in milking (DIM) were selected from 94 cows. Based on the milk yield of the cows, they were divided into two groups of nine cows each, the high milk yield group (HP) (44.57 ± 2.11 kg/day) and the low milk yield group (LP) (26.71 ± 0.70 kg/day). The experimental cows were fed the same diet and kept under the same management system for more than 60 days. Rumen metagenomics revealed that two Archaea genera, one Bacteria genus, eight Eukaryota genera, and two Virus genera differ between the HP and LP groups. The analysis of metabolites in the rumen fluid, milk, and serum showed that several metabolites differed between the HP and LP groups. Correlation analysis between the predominant microbiota and milk yield-associated metabolites (MP-metabolites) revealed that four Bacteria and two Eukaryota genera have a positive relationship with MP-metabolites. Pathway enrichment analysis of the differential metabolites revealed that five pathways were enriched in all the samples (two pathways in the milk, two pathways in the serum, and one pathway in the rumen fluid). Further investigation revealed that the low milk yield observed in the LP group might be due to an upregulation in dopamine levels in the rumen fluid and milk, which could inhibit the release of prolactin or suppress the action of oxytocin in the udder resulting in reduced milk yield. On the other hand, the high milk yield in the HP group is attributed to an upregulation in citrulline, and N-acetylornithine, which could be used as substrates for energy metabolism in the citric acid cycle and ultimately gluconeogenesis.
Design of Pb(II)-Specific E. coli-Based Biosensors by Engineering Regulatory Proteins and Host Cells
Authors: Jeon Yangwon , Lee Yejin , Jang Geupil , Kim Bong-Gyu , Yoon Youngdae
Published: May 2022

Bacterial cell-based biosensors have been widely developed for detecting environmental toxic materials. The znt-operon in Escherichia coli is a Zn(II)-responsive genetic system and is employed in Zn(II), Cd(II), and Hg(II)-sensing biosensors. In this study, point mutations were introduced in the regulatory protein ZntR to modulate its target selectivity, and metal ion-exporting genes, such as copA and zntA, in host cells were deleted to increase cellular metal ion levels and enhance specificity. Thus, the overall responses of the E. coli cell-based biosensors toward metal(loid) ions were increased, and their selectivity, which was originally for Cd(II) and Hg(II), was shifted to Pb(II). The gene encoding ZntA, known as the Zn(II)-translocating P-type ATPase, showed an impact on the ability of E. coli to export Pb(II), whereas copA deletion showed no significant impact. Noteworthily, the newly generated biosensors employing ZntR Cys115Ile showed the capacity to detect under 5 nM Pb(II) in solution, without response to other tested metal ions within 0–100 nM. To understand the marked effect of single point mutations on ZntR, computational modeling was employed. Although it did not provide clear answers, changes in the sequences of the metal-binding loops of ZntR modulated its transcriptional strength and target selectivity. In summary, the approaches proposed in this study can be valuable to generate new target-sensing biosensors with superior selectivity and specificity, which can in turn broaden the applicability of cell-based biosensors to monitor Pb(II) in environmental systems.
Viola betonicifolia-Mediated Biosynthesis of Silver Nanoparticles for Improved Biomedical Applications
Authors: Jang Yingping , Zhang Xiaoya , Zhu Rongxue , Li Songlin , Sun Shiyu , Li Wenqiang , Liu Hao
Published: May 2022

We report the biosynthesis of silver (Ag) nanoparticles (NAPs) (LEVB-Ag NAPs) by an environmentally friendly green synthesis approach using the phytoconstituents of Viola betonicifolia leaf extract. The spectroscopic techniques were employed to characterize biosynthesized LEVB-Ag NAPs successfully. Biosynthesized LEVB-Ag NAPs were assessed for antibacterial and antimycotic activities against bacterium and mycological strains (H. pylori, S. epidermidis, C. tropicalis, and T. rubrum) using the serial dilution method. They were also evaluated for their biofilm inhibiting potential against both bacterial and fungi species. They were further assessed for the cytobiocompatible potential with two normal cell lines (293T and hMSC). The results demonstrate that the biosynthesized LEVB-Ag NAPs showed superior log10 reduction in bacterial and fungal growth and presented more than 99.50% killing efficiency. Moreover, biosynthesized LEVB-Ag NAPs excellently inhibited the biofilm formation of bacterial (Gram-positive and Gram-negative) and mycological strains and presented more than 80% biofilm inhibiting percentage compared to both plant extract and CHE-Ag NAPs. They further presented good cytobiocompatibility in vitro with 293T and hMSC cells compared to CHE-Ag NAPs. Biosynthesized LEVB-Ag NAPs presented superior antibacterial, antimycotic, biofilm inhibition, and cytobiocompatible results that might be attributed to the synergistic effect of the NAPs’ physiochemical properties and the immobilized phytoconstituents from plant leaf extract on their surface. Hence, biosynthesized LEVB-Ag NAPs may be a promising contender for a variety of therapeutic applications.
Highly Efficient Methods to Culture Mouse Cholangiocytes and Small Intestine Organoids
Authors: Chen Wenyi , Yao Qigu , Wang Ruo , Fen Bing , Chen Junyao , Xu Yanping , Yu Jiong , Li Lanjuan , Cao Hongcui
Published: May 2022

Background

Organoids, which enable disease modeling and drug screening closer to an in vivo environment, can be isolated and grown from organs such as the brain, small intestine, kidney, lungs, and liver. To facilitate the establishment of liver and small intestinal organoids, we developed efficient protocols for cholangiocytes and intestine crypts collecting and organoid culturing.

Methods

Cholangiocytes were collected from intrahepatic bile ducts, the gallbladder, and small intestine crypts by gravity settling and multistep centrifugation methods. The cells isolated were embedded with Matrigel and grew in three-dimensional spheroids in a suitable culture medium. The stability of organoid cells was assessed by subculture, cryopreservation, and thawing. RNA and DNA extraction of organoids, as well as immunostaining procedure, were also optimized. Hand-picking procedures were developed and performed to ensure similar growth characteristics of organoids.

Results

A large number of cholangiocytes and small intestine crypts were collected under these protocols. Cholangiocytes developed into cyst-like structures after 3–4 days in Matrigel. After 1–2 weeks of cultivation, small intestinal organoids (in-orgs) developed buds and formed a mature structure. Compared to organoids derived from the gallbladder, cholangiocyte organoids (Cho-orgs) from intrahepatic the bile ducts grew more slowly but had a longer culture term, expressed the cholangiocytes markers Krt19 and Krt7, and recapitulated in vivo tissue organization.

Conclusions

Our protocols simplified the cell collection procedure and avoided the possibility of exposing tissue-derived stem cells to mechanical damage or chemical injury by gravity settling and multistep centrifugation. In addition, our approach allowed similar growth characteristics of organoids from different mammalian tissue sources. The protocol requires 2–4 weeks to establish a stable organoid growth system. Organoids could be stably passaged, cryopreserved, and recovered under protocol guidance. Besides, the organoids of cholangiocytes and small intestines retained their original tissue characteristics, such as tissue-specific marker expression, which prepares them for further experiments such as preclinical in vitro trials and mechanism research studies.
SMARCC1 Enters the Nucleus via KPNA2 and Plays an Oncogenic Role in Bladder Cancer
Authors: Wei Zhengmao , Xu Jinming , Li Weiqing , Ou Longhua , Zhou Yingchen , Wang Yan , Shi Bentao
Published: May 2022

Background: SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin subfamily C member 1 (SMARCC1), a component of the SWI/SNF complex, is thought to be an oncogene in several kinds of cancer.

Materials and methods: The potential interaction between SMARCC1 and KPNA2 was inquired by Spearman’s correlation analysis, immunofluorescence staining and co-immunoprecipitation (Co-IP) assays. The immunohistochemistry staining, RT-PCR and western blot assay were taken for determining the expression levels of SMARCC1. And CCK-8, transwell assay, cell apoptosis assay, cell cycle analysis and subcutaneous tumor model were conducted to explore the role of SMARCC1 in carcinogenesis of bladder cancer.

Results: In our experiments, Spearman’s correlation analysis, immunofluorescence staining and co-immunoprecipitation (Co-IP) assays showed that SMARCC1 interacted with KPNA2, and after knockdown of KPNA2, Nup50 and Nup153, the nuclear content of SMARCC1 decreased while the amount of SMARCC1 protein remaining in the cytoplasm increased, indicating that SMARCC1 could be transported into the nucleus via KPNA2 and thus acted as an oncogene. We found that both the mRNA and protein expression levels of SMARCC1 were increased in bladder cancer, and increased SMARCC1 expression was significantly associated with a higher T stage and poorer prognosis in bladder cancer patients. Knockdown of SMARCC1 slowed the growth of the two tested cell lines and clearly arrested the cell cycle at the G0/G1 phase checkpoint. Moreover, the migratory ability was significantly decreased and the number of apoptotic cells was increased.

Conclusion: On the whole, our results demonstrate KPNA2, Nup50 and Nup153 regulate the process of SMARCC1 nuclear translocation in BC. SMARCC1 may be a competent candidate as a diagnostic and therapeutic target for BC. Further studies are required to research the mechanism and assess the role of SMARCC1 in vivo.