Bladder-cancer-derived exosomal circRNA_0013936 promotes suppressive immunity by up-regulating fatty acid transporter protein 2 and down-regulating receptor-interacting protein kinase 3 in PMN-MDSCs.

BACKGROUND: Polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) is one of the causes of tumor immune tolerance and failure of cancer immunotherapy. Here, we found that bladder cancer (BCa)-derived exosomal circRNA_0013936 could enhance the immunosuppressive activity of PMN-MDSCs by regulating the expression of fatty acid transporter protein 2 (FATP2) and receptor-interacting protein kinase 3 (RIPK3). However, the underlying mechanism remains largely unknown. METHODS: BCa-derived exosomes was isolated and used for a series of experiments. RNA sequencing was used to identify the differentially expressed circRNAs. Western blotting, immunohistochemistry, immunofluorescence, qRT-PCR, ELISA and Flow cytometry were performed to reveal the potential mechanism of circRNA_0013936 promoting the immunosuppressive activity of PMN-MDSC. RESULTS: CircRNA_0013936 enriched in BCa-derived exosomes could promote the expression of FATP2 and inhibit the expression of RIPK3 in PMN-MDSCs. Mechanistically, circRNA_0013936 promoted the expression of FATP2 and inhibited the expression of RIPK3 expression via sponging miR-320a and miR-301b, which directly targeted JAK2 and CREB1 respectively. Ultimately, circRNA_0013936 significantly inhibited the functions of CD8+ T cells by up-regulating FATP2 through the circRNA_0013936/miR-320a/JAK2 pathway, and down-regulating RIPK3 through the circRNA_0013936/miR-301b/CREB1 pathway in PMN-MDSCs. CONCLUSIONS: BCa-derived exosomal circRNA_0013936 promotes suppressive immunity by up-regulating FATP2 through the circRNA_0013936/miR-320a/JAK2 pathway and down-regulating RIPK3 through the circRNA_0013936/miR-301b-3p/CREB1 pathway in PMN-MDSCs. These findings help to find new targets for clinical treatment of human bladder cancer.

Development and experimental validation of an M2 macrophage and platelet-associated gene signature to predict prognosis and immunotherapy sensitivity in bladder cancer.

Platelets and M2 macrophages both play crucial roles in tumorigenesis, but their relationship and the prognosis value of the relative genes in bladder cancer (BLCA) remain obscure. In the present study, we found that platelets stimulated by BLCA cell lines could promote M2 macrophage polarization, and platelets were significantly associated with the infiltration of M2 macrophages in BLCA samples. Through the bioinformatic analyses, A2M, TGFB3, and MYLK, which were associated with platelets and M2 macrophages, were identified and verified in vitro and then included in the predictive model. A platelet and M2 macrophage-related gene signature was constructed to evaluate the prognosis and immunotherapeutic sensitivity, helping to guide personalized treatment and to disclose the underlying mechanisms.

Population genetic diversity and environmental adaptation of Tamarix hispida in the Tarim Basin, arid Northwestern China.

Arid ecosystems, characterized by severe water scarcity, play a crucial role in preserving Earth's biodiversity and resources. The Tarim Basin in Northwestern China, a typical arid region isolated by the Tianshan Mountains and expansive deserts, provides a special study area for investigating how plant response and adaptation to such environments. Tamarix hispida, a species well adapted to saline-alkaline and drought conditions, dominates in the saline-alkali lands of the Tarim Basin. This study aims to examine the genetic diversity and environmental adaptation of T. hispida in the Tarim Basin. Genomic SNPs for a total of 160 individuals from 17 populations were generated using dd-RAD sequencing approach. Population genetic structure and genetic diversity were analyzed by methods including ADMIXTURE, PCA, and phylogenetic tree. Environmental association analysis (EAA) was performed using LFMM and RDA analyses. The results revealed two major genetic lineages with geographical substitution patterns from west to east, indicating significant gene flow and hybridization. Environmental factors such as Precipitation Seasonality (bio15) and Topsoil Sand Fraction (T_SAND) significantly shaped allele frequencies, supporting the species' genetic adaptability. Several genes associated with environmental adaptation were identified and annotated, highlighting physiological and metabolic processes crucial for survival in arid conditions. The study highlights the role of geographical isolation and environmental factors in shaping genetic structure and adaptive evolution. The identified adaptive genes related to stress tolerance emphasize the species' resilience and highlight the importance of specific physiological and metabolic pathways.

Exploring the link between dietary omega-3 and omega-6 fatty acid intake and rheumatoid arthritis risk: NHANES 1999-2020 study.

OBJECTIVES: The association between the ingestion of n-3 and n-6 fatty acids and rheumatoid arthritis (RA) remains unclear. To address this, this study utilised data from the National Health and Nutrition Examination Survey (NHANES) spanning from 1999 to 2020. METHODS: Dietary intake information on n-3 and n-6 fatty acids was gathered through 24-hour interviews about dietary recall and adjusted based on weight. RA patient data was collected using questionnaires. Associations were evaluated using logistic regression and spline analyses. The study included a total of 50,352 participants in a cross-sectional manner. RESULTS: In the adjusted Model 2, higher odds ratios (ORs) of 0.72 (95% CI: 0.60-0.86) and 0.76 (95% CI: 0.62-0.92) were observed for n-3 and n-6 fatty acid intake, respectively, compared to the lowest category. CONCLUSIONS: The results suggest a negative correlation between the ingestion of n-3 and n-6 fatty acids and the risk of rheumatoid arthritis in US adults.

Pursuing sustainable productivity with millions of smallholder farmers.

Sustainably feeding a growing population is a grand challenge, and one that is particularly difficult in regions that are dominated by smallholder farming. Despite local successes, mobilizing vast smallholder communities with science- and evidence-based management practices to simultaneously address production and pollution problems has been infeasible. Here we report the outcome of concerted efforts in engaging millions of Chinese smallholder farmers to adopt enhanced management practices for greater yield and environmental performance. First, we conducted field trials across China's major agroecological zones to develop locally applicable recommendations using a comprehensive decision-support program. Engaging farmers to adopt those recommendations involved the collaboration of a core network of 1,152 researchers with numerous extension agents and agribusiness personnel. From 2005 to 2015, about 20.9 million farmers in 452 counties adopted enhanced management practices in fields with a total of 37.7 million cumulative hectares over the years. Average yields (maize, rice and wheat) increased by 10.8-11.5%, generating a net grain output of 33 million tonnes (Mt). At the same time, application of nitrogen decreased by 14.7-18.1%, saving 1.2 Mt of nitrogen fertilizers. The increased grain output and decreased nitrogen fertilizer use were equivalent to US$12.2 billion. Estimated reactive nitrogen losses averaged 4.5-4.7 kg nitrogen per Megagram (Mg) with the intervention compared to 6.0-6.4 kg nitrogen per Mg without. Greenhouse gas emissions were 328 kg, 812 kg and 434 kg CO2 equivalent per Mg of maize, rice and wheat produced, respectively, compared to 422 kg, 941 kg and 549 kg CO2 equivalent per Mg without the intervention. On the basis of a large-scale survey (8.6 million farmer participants) and scenario analyses, we further demonstrate the potential impacts of implementing the enhanced management practices on China's food security and sustainability outlook.

Ultrastable Graphite-Potassium Anode through Binder Chemistry.

Graphite with abundant reserves has attracted enormous research interest as an anode of potassium-ion batteries (PIBs) owing to its high plateau capacity of 279 mAh g-1 at ≈0.2 V in conventional carbonate electrolytes. Unfortunately, it suffers from fast capacity decay during K+ storage. Herein, an ultrastable graphite-potassium anode is developed through binder chemistry. Polyvinyl alcohol (PVA) is utilized as a water-soluble binder to generate a uniform and robust KF-rich SEI film on the graphite surface, which can not only inhibit the electrolyte decomposition, but also withstand large volume expansion during K+ -insertion. Compared to the PVDF as binder, PVA-based graphite anode can operate for over 2000 cycles (running time of 406 days at C/3) with 97% capacity retention in KPF6 -based electrolytes. The initial Coulombic efficiency (ICE) of graphite anode is as high as 81.6% using PVA as the binder, higher than that of PVDF (40.1%). Benefiting from the strong adhesion ability of PVA, a graphite||fluorophosphate K-ion full battery is further built through 3D printing, which achieves a record-high areal energy of 8.9 mWh cm-2 at a total mass loading of 38 mg cm-2 . These results demonstrate the important role of binder in developing high-performance PIBs.

A multicenter noninferior randomized controlled study of sentinel lymph node biopsy alone versus sentinel lymph node biopsy plus lymphadenectomy for patients with stage I endometrial cancer, INSEC trial concept.

BACKGROUND: Up to the present time, there has remained a lack of strong evidence as to whether sentinel lymph node biopsy can replace lymphadenectomy for early endometrial cancer. The traditional surgery for endometrial cancer includes pelvic lymphadenectomy and paraaortic lymph node resection, but complications often seriously affect patients' quality of life. Two randomized controlled trials with large samples have proved that lymphadenectomy does not improve the overall recurrence rate and survival rate of patients. On the contrary, it increases the incidence of complications and even mortality. The current trial is designed to clarify whether sentinel lymph node biopsy can replace lymphadenectomy for early endometrial cancer patients with negative lymph nodes. METHODS: This study is a randomized, open-label, multicenter and non-inferiority controlled clinical trial in China. Potential participants will be patients with pathologically confirmed endometrial cancer at the Zhejiang Cancer Hospital, Jiaxing Maternity and Child Health Care Hospital, and the First Hospital of Jiaxing in China. The total sample size for this study is 722. Patients will be randomly assigned in a 1:1 ratio to two groups. Patients in one group will undergo sentinel lymph node biopsy + total hysterectomy + bilateral salpingo-oophorectomy ± paraaortic lymph node resection. Patients in the other group will undergo sentinel lymph node biopsy + total hysterectomy + bilateral salpingo-oophorectomy + pelvic lymphadenectomy ± paraaortic lymph node resection. The 3-year disease-free survival rate, overall survival rate, quality of life (use EORTC QLQ-C30 + QLQ-CX24), and perioperative related indexes of the two groups will be compared. RESULTS: We expect to find that for patients with early endometrial cancer, the 3-year disease-free survival rate following sentinel lymph node biopsy with indocyanine green combined with near-infrared fluorescence imaging is similar to that following lymphadenectomy. The operation time, as well as incidence of pelvic lymphocyst, lymphedema of lower limb, and edema of vulva in patients who only undergo sentinel lymph node biopsy are expected to be significantly lower than in patients who undergo lymphadenectomy. The quality of life of patients who undergo sentinel lymph node biopsy alone will be significantly better than that of patients who undergo lymph node dissection. CONCLUSION: This will prove that the prognosis of sentinel lymph node biopsy alone with indocyanine green combined with near-infrared fluorescence imaging is not inferior to that of sentinel lymph node biopsy plus lymphadenectomy for early stage endometrial cancer with negative nodal assessment intraoperatively. In addition, sentinel lymph node biopsy alone with indocyanine green combined with near-infrared fluorescence imaging results in fewer surgical complications and gives patients better quality of life. TRIAL REGISTRATION: chictr.org.cn, ChiCTR1900023161. Registered 14 May 2019, http://www.chictr.org.cn/edit.aspx?pid=38659&htm=4 .

Optimistic contributions of plant growth-promoting bacteria for sustainable agriculture and climate stress alleviation.

Global climate change is the major cause of abiotic and biotic stresses that have adverse effects on agricultural productivity to an irreversible level, thus threatening to limit gains in production and imperil sustainable agriculture. These climate change-induced abiotic stresses, especially saline, drought, extreme temperature, and so on affect plant morphological, physiological, biochemical, and metabolic characteristics through various pathways and mechanisms, ultimately hindering plant growth, development, and productivity. However, overuse and other inappropriate uses of agrochemicals are not conducive to the protection of natural resources and the environment, thus hampering sustainable agricultural development. With the vigorous development of modern agriculture, the application of plant growth-promoting bacteria (PGPB) can better ensure sustainable agriculture, due to their ability to improve soil properties and confer stress tolerance in plants. This review deciphered the underlying mechanisms of PGPB involved in enhancing plant stress tolerance and performance under various abiotic and biotic stresses. Moreover, the recent advancements in PGPB inoculation techniques, the commercialization of PGPB-based technology and the current applications of PGPB in sustainable agriculture were extensively discussed. Finally, an outlook on the future directions of microbe-aided agriculture was pointed out. Providing insights into plant-PGPB interactions under biotic and abiotic stresses and offering evidence and strategies for PGPB better commercialization and implementation can inspire the development of innovative solutions exploiting PGPB under climatological conditions.

Nitrogen in the Yangtze River Basin: Pollution Reduction through Coupling Crop and Livestock Production.

Livestock production poses a threat to water quality worldwide. A better understanding of the contribution of individual livestock species to nitrogen (N) pollution in rivers is essential to improve water quality. This paper aims to quantify inputs of dissolved inorganic nitrogen (DIN) to the Yangtze River from different livestock species at multiple scales and explore ways for reducing these inputs through coupling crop and livestock production. We extended the previously developed model MARINA (Model to Assess River Input of Nutrient to seAs) with the NUFER (Nutrient flows in Food chains, Environment, and Resource use) approach for livestock. Results show that DIN inputs to the Yangtze River vary across basins, sub-basins, and 0.5° grids, as well as across livestock species. In 2012, livestock production resulted in 2000 Gg of DIN inputs to the Yangtze River. Pig production was responsible for 55-85% of manure-related DIN inputs. Rivers in the downstream sub-basin received higher manure-related DIN inputs than rivers in the other sub-basins. Around 20% of the Yangtze basin is considered as a manure-related hotspot of river pollution. Recycling manure on cropland can avoid direct discharges of manure from pig production and thus reduce river pollution. The potential for recycling manure is larger in cereal production than in other crop species. Our results can help to identify effective solutions for coupling crop and livestock production in the Yangtze basin.

Feedback loop between fatty acid transport protein 2 and receptor interacting protein 3 pathways promotes polymorphonuclear neutrophil myeloid-derived suppressor cells-potentiated suppressive immunity in bladder cancer.

BACKGROUND: Polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) promote tumor immune tolerance and cause tumor immunotherapy failure. In this study, we found that high PMN-MDSCs infiltration, overexpressed fatty acid transporter protein 2 (FATP2) and underexpressed receptor-interacting protein kinase 3 (RIPK3) existed in the mouse and human bladder cancer tissues. However, the related mechanisms remain largely unknown. METHODS AND RESULTS: Both FATP2 and RIPK3 expressions were associated with clinical stage. FATP2 knockout or up-regulating RIPK3 reduced the synthesis of prostaglandin E2 (PGE2) in PMN-MDSCs, attenuated the suppressive activity of PMN-MDSCs on CD8+ T cells functions and inhibited the tumor growth. There was a PGE2-mediated feedback loop between FATP2 and RIPK3 pathways, which markedly promoted the immunosuppressive activity of PMN-MDSCs. Combination therapy with inhibition of FATP2 and activation of RIPK3 can effectively inhibit tumor growth. CONCLUSIONS: This study demonstrated that a feedback loop between FATP2 and RIPK3 pathways in PMN-MDSCs significantly promoted the synthesis of PGE2, which severely impaired the CD8+ T cell functions. This study may provide new ideas for immunotherapy of human bladder cancer.

Association of Hypertension with Knee Pain Severity Among People with Knee Osteoarthritis.

PURPOSE: To examine the association of hypertension with knee pain severity in individuals with knee osteoarthritis (OA). DESIGN: Cross-sectional study of baseline data collected by the Osteoarthritis Initiative. METHODS: Participants with knee OA (N=1,363) were categorized into four groups based on blood pressure (BP): 1) systolic < 120 mm HG and diastolic < 80 mm Hg; 2) 120 ≤ systolic < 130 mm Hg and diastolic < 80 mm Hg; 3) 130 ≤ systolic < 140 mm Hg or 80 ≤ diastolic < 90 mm Hg; 4) systolic ≥ 140 mm Hg or diastolic ≥ 90 mm Hg. OA knee pain severity was measured by Pain subscale of Western Ontario and McMaster Universities Osteoarthritis Index in the past 48 hours, Pain subscale of Knee Injury and Osteoarthritis Outcome Score (KOOS) in the past 7 days, and numeric rating scale (NRS) in the past 30 days. Linear regression was used to examine the relationship between hypertension and knee pain severity. RESULTS: Compared with the normal BP group, individuals with stage 2 hypertension reported significantly higher OA knee pain severity by KOOS in the past 7 days (ß=-2.05 [95% CI -4.09, -0.01], p=0.049) and by NRS in the past 30 days (ß=0.31 [95% CI 0.01, 0.62], p=0.045) after adjustments for demographic and medical factors. CONCLUSIONS: Hypertension was associated with higher OA knee pain severity in individuals with knee OA. CLINICAL IMPLICATIONS: Nurses can recommend adjunctive non-pharmacological treatments and adherence strategies to help control hypertension, which may help decrease OA knee pain.

The relationship between FOSB and SOCS3 gene polymorphisms and the susceptibility to periodontitis and osteopenia in the Chinese population.

The aim is to analyze the relationship and significance of the FOS, FOSB, Suppressors of cytokine signaling (SOCS), and hypoxia-inducible factor 1 (HIF1) gene loci and their polymorphisms with periodontitis and osteopenia in the Chinese population. In this case-control study, data on questionnaires, periodontal examination, bone mineral density, and FOS, FOSB, SOCS3, and HIF1 gene loci and their polymorphisms were obtained from 474 participants. The data were analyzed using the analysis of variance, Kruskal-Wallis test, χ2 test, and logistic regression. The incidence of osteopenia was significantly increased in patients with periodontitis compared to controls (58.6 vs. 34.4%, P < 0.001). Accordingly, the risk was increased 2.21-fold compared with controls (95% CI 2.09-4.95). Osteopenia patients had a significantly higher risk of periodontitis than patients with normal bone density (OR = 3.22, 95% CI 2.09-4.94). There were significant positive associations between FOSB and SOCS3 polymorphisms and periodontitis and osteopenia susceptibility. Individuals carrying the G/G genotype of the FOSB gene rs708905 locus had an increased risk of periodontitis (OR = 5.06, 95% CI 2.36-10.86) and osteopenia (OR = 3.26, 95% CI 1.34-7.96). Compared with the C/C genotype, the A/A genotype of the FOSB rs8105114 locus was associated with a significantly higher risk of periodontitis (OR = 2.14, 95% CI 1.02-4.53) and osteopenia (OR = 2.85, 95% CI 1.12-7.22). Compared with the A/A genotype, the risk of periodontitis in the G/G genotype of the SOCS3 rs7207782 locus was increased 3.10-fold (P < 0.001), and the risk of osteopenia was increased 2.01-fold (P = 0.023). There was a bidirectional relationship between periodontitis and osteopenia. The rs708905 G/G and rs8105114 A/A genotypes of FOSB and the rs7207782 G/G genotype of SOCS3 were risk factors for both periodontitis and osteopenia in the Chinese population, which could increase knowledge about disease­specific and cross­disease genetic pattern.

[Preliminary Study on the Effect of Whole-Process Case Management Based on Service Process Design on the Function and Satisfaction of Patients Undergoing Total Knee Arthroplasty].

Objective: To investigate the effect of whole-process case management based on service process design on patients undergoing total knee arthroplasty (TKA) in areas including pain, function, satisfaction, and complications. Methods: A total of 204 patients who underwent unilateral TKA between April 2021 and March 2022 at the Department of Orthopedics, West China Hospital, Sichuan University were enrolled. By using a random number table, the patients were randomly assigned to two groups, 102 in the general case management group (group G) and 102 in the whole-process case management group (group W). Patients in group G received traditional perioperative case management, while those in the whole-process case management group received integrated case management optimized on the basis of the service process design. The two groups of patients were studied through comparison of their general data, Visual Analogue Scale (VAS) pain score, knee flexion and range of motion, Hospital for Special Surgery (HSS) knee score, the 18-item Patient Satisfaction Questionnaire Short Form (PSQ-18), ability to climb stairs, and complications at 3 days and 3, 8, and 12 weeks after TKA. Results: There was no significant difference between the two groups in patient general information or baseline data collected at the time of enrollment ( P>0.05). There was no significant difference in HSS score, joint range of motion, and VAS pain score between the two groups before the surgery and 3 days after the surgery ( P>0.05). However, the HSS score, joint range of motion, and VAS pain scores of group W were significantly superior to those of group G at 3, 8 and 12 weeks after the surgery (all P<0.05). In addition, group W demonstrated significantly better ability to climb up and down stairs than that of group G at 12 weeks after the surgery ( P< 0.001). In terms of satisfaction, patients in group W were significantly more satisfied than those in group G at 3 days, and 3, 8, and 12 weeks after the surgery ( P<0.001). Conclusion: Whole-process case management based on service process design has a positive effect of relieving pain, increasing range of motion, improving function, increasing satisfaction, and reducing complications in patients undergoing TKA.

Involvement of a FAD-Linked Oxidase RSc0454 for Expression of the Type III Secretion System and Pathogenicity in Ralstonia solanacearum.

Ralstonia solanacearum RSc0454 is predicted as a FAD-linked oxidase based on protein homologies, while it contains distinct domains of lactate dehydrogenase and succinate dehydrogenase. A previous study demonstrated that RSc0454 exhibits lactate dehydrogenase activity using pyruvate and NADH as substrates, and is essential for pathogenicity of R. solanacearum. Here, we genetically characterized involvement of RSc0454 on bacterial growth and expression of genes for the type III secretion system (T3SS, a pathogenicity determinant) in R. solanacearum. The RSc0454 mutant grew normally in rich medium but grew faintly in host plants, and failed to grow in minimal medium. Supplementary succinate but not lactate could substantially restore some phenotypes of RSc0454 mutants, including faint growth in host plants, diminished growth in the minimal medium, and lost pathogenicity toward host plants. Expression of T3SS genes is directly controlled by a master regulator, HrpB, and hrpB expression is positively regulated by HrpG and PrhG in parallel ways. Deletion of RSc0454 substantially reduced expression levels of hrpB and T3SS both in vitro and in planta. Moreover, RSc0454 is revealed to be required for the T3SS expression via HrpG and PrhG, although through some novel pathway, and impaired expression of these genes was not due to growth deficiency of RSc0454 mutants. RSc0454 is suggested to be important for redox balance inside cells, and supplementary NADH partially restored diminished growth of the RSc0454 mutant in the minimal medium only in the presence of succinate at some moderate concentrations, indicating that the unbalanced redox in the RSc0454 mutant might be responsible for its diminished growth in the minimal medium. Taken together, these results provide novel insights into the understanding of various biological functions of this FAD-linked oxidase RSc0454 and involvement of the redox balance on expression of the T3SS in R. solanacearum.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Resolving Membrane Protein-Protein Interactions in Live Cells with Pulsed Interleaved Excitation Fluorescence Cross-Correlation Spectroscopy.

The cell plasma membrane (PM) contains thousands of proteins that sense and respond to the outside environment. These proteins have evolved sensitivity to a wide variety of physical and chemical signals and act as a delivery system across the PM. Membrane proteins are critical for information flow and decision making in the cell and thus are important targets in drug development. A critical aspect of membrane protein function is the way they interact with other proteins, often through the formation of dimers or small oligomers that regulate function at the protein, cell, and organism levels. Resolving membrane protein interactions in a live cell environment is challenging because of the chemical diversity and spatial heterogeneity of the PM. In this Account, we describe a fluorescence technique called pulsed interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS) that is ideally suited to quantify membrane associations in live cells. PIE-FCCS is a two-color fluorescence fluctuation method that can simultaneously measure the concentration, mobility, proximity, and oligomerization state of membrane proteins in situ. It has several advantages over two related approaches, single-molecule tracking (SMT) and Förster resonance energy transfer (FRET), including that it measures all of the properties listed above in a single measurement. Another advantage is that PIE-FCCS is most sensitive at the physiological expression levels for many membrane proteins rather than the very low or high levels typical in other techniques. Here, we review the history of FCCS as it has been applied to study membrane protein interactions in cells. We also describe PIE-FCCS and the advantages it has over biochemical approaches like coimmunoprecipitation (co-IP) and proximity ligation assays (PLA). Finally, we review two classes of membrane proteins that have been studied with FCCS and PIE-FCCS: receptor tyrosine kinases (RTKs) and G protein-coupled receptors (GPCRs). For RTKs, ligand induced dimerization directly regulates the catalytic activity of the kinase, but higher order oligomerization and ligand-independent dimerization can complicate this historically simple paradigm. PIE-FCCS data have resolved a low population of EGFR dimers under basal conditions and assembly into multimers when stimulated with ligand. While GPCRs function primarily as monomers, dimerization has been hypothesized to regulate function for some receptors. PIE-FCCS data have established the dimerization potential of rhodopsin at low densities and were critical for the discovery of a novel dimerization interface in human cone opsins. This Account describes the how FCCS and PIE-FCCS can reveal the details of quaternary interactions in each of these receptor systems.

Urinary Exosomal miRNAs as biomarkers of bladder Cancer and experimental verification of mechanism of miR-93-5p in bladder Cancer.

BACKGROUND: Bladder cancer (BC) is one of the most common malignancies globally. Early diagnosis of it can significantly improve patients' survival and quality of life. Urinary exosomes (UEs)-derived miRNAs might be a promising biomarker for BC detection. METHOD: A total of 12 patients with BC and 4 non-cancerous participants (as healthy control) were recruited from a single center between March 2018 and December 2019 as the discovery set. Midstream urine samples from each participants were collected and high-throughput sequencing and differentially expression analysis were conducted. Combined with miRNA expression profile of BC tissue from The Cancer Genome Atlas (TCGA), miRNAs biomarkers for BC were determined. Candidate miRNAs as biomarkers were selected followed by verification with a quantitative reverse-transcription polymerase chain reaction assay in an independent validation cohort consisting of 53 BC patients and 51 healthy controls. The receiver-operating characteristic (ROC) curve was established to evaluate the diagnostic performance of UE-derived miRNAs. The possible mechanism of miRNAs were revealed by bioinformatic analysis and explored in vitro experiments. RESULTS: We identified that miR-93-5p, miR-516a-5p were simultaneously significantly increased both in UEs from BC compared with healthy control and BC tissue compared with normal tissue, which were verified by RT-qPCR in the validation cohort. Subsequently, the performance to discover BC of the miR-93-5p, miR-516a-5p was further verified with an area under ROC curve (AUC) of 0.838 and 0.790, respectively, which was significantly higher than that of urine cytology (AUC = 0.630). Moreover, miR-93-5p was significantly increased in muscle-invasive BC compared with non-muscle-invasive BC with an AUC of 0.769. Bioinformatic analysis revealed that B-cell translocation gene 2(BTG2) gene may be the hub target gene of miR-93-5p. In vitro experiments verified that miR-93-5p suppressed BTG2 expression and promoted BC cells proliferation, invasion and migration. CONCLUSION: Urine derived exosomes have a distinct miRNA profile in BC patients, and urinary exosomal miRNAs could be used as a promising non-invasive tool to detect BC. In vitro experiments suggested that miR-93-5p overexpression may contribute to BC progression via suppressing BTG2 expression.

Adaboost-Based Machine Learning Improved the Modeling Robust and Estimation Accuracy of Pear Leaf Nitrogen Concentration by In-Field VIS-NIR Spectroscopy.

Different cultivars of pear trees are often planted in one orchard to enhance yield for its gametophytic self-incompatibility. Therefore, an accurate and robust modelling method is needed for the non-destructive determination of leaf nitrogen (N) concentration in pear orchards with mixed cultivars. This study proposes a new technique based on in-field visible-near infrared (VIS-NIR) spectroscopy and the Adaboost algorithm initiated with machine learning methods. The performance was evaluated by estimating leaf N concentration for a total of 1285 samples from different cultivars, growth regions, and tree ages and compared with traditional techniques, including vegetation indices, partial least squares regression, singular support vector regression (SVR) and neural networks (NN). The results demonstrated that the leaf reflectance responded to the leaf nitrogen concentration were more sensitive to the types of cultivars than to the different growing regions and tree ages. Moreover, the AdaBoost.RT-BP had the best accuracy in both the training (R2 = 0.96, root mean relative error (RMSE) = 1.03 g kg-1) and the test datasets (R2 = 0.91, RMSE = 1.29 g kg-1), and was the most robust in repeated experiments. This study provides a new insight for monitoring the status of pear trees by the in-field VIS-NIR spectroscopy for better N managements in heterogeneous pear orchards.

Older adults' social network and support and its association with physical activity.

The purpose of this study was to describe older adults' social network and support during a physical activity intervention and its association with physical activity. Mixed methods were used for this secondary analysis of existing data from a physical activity intervention. Seventy-three participants who completed a session by telephone on using support comprised the sample. Participants reported on average approximately eight individuals in their social network. Half reported support for physical activity from a spouse/partner, adult child, or friend. Those who perceived support for self-reported physical activity tended to have greater improvements on average from baseline for mean daily minutes of physical activity than those with no perceived support. Four-fifths developed a plan for a spouse/partner, friend, adult child, or sibling to provide physical activity support. Participants reported that the session was helpful and increased their confidence in using support; over one-quarter intended to use support after the intervention ended.

Functional Characterization of RsRsgA for Ribosome Biosynthesis and Expression of the Type III Secretion System in Ralstonia solanacearum.

RsgA plays an important role in maturation of 30S subunit in many bacteria that assists in the release of RbfA from the 30S subunit during a late stage of ribosome biosynthesis. Here, we genetically characterized functional roles of RsgA in Ralstonia solanacearum, hereafter designated RsRsgA. Deletion of R. solanacearum rsgA or rbfA resulted in distinct deficiency of 16S ribosomal RNA, significantly slowed growth in broth medium, and diminished growth in nutrient-limited medium, which are similar as phenotypes of rsgA mutants and rbfA mutants of Escherichia coli and other bacteria. Our gene-expression studies revealed that RsRsgA is important for expression of genes encoding the type III secretion system (T3SS) (a pathogenicity determinant of R. solanacearum) both in vitro and in planta. Compared with the wild-type R. solanacearum strain, proliferation of the rsgA and rbfA mutants in tobacco leaves was significantly impaired, while they failed to migrate into tobacco xylem vessels from infiltrated leaves, and hence, these two mutants failed to cause any bacterial wilt disease in tobacco plants. It was further revealed that rsgA expression was highly enhanced under nutrient-limited conditions compared with that in broth medium and RsRsgA affects T3SS expression through the PrhN-PrhG-HrpB pathway. Moreover, expression of a subset of type III effectors was substantially impaired in the rsgA mutant, some of which are responsible for R. solanacearum GMI1000 elicitation of a hypersensitive response (HR) in tobacco leaves, while RsRsgA is not required for HR elicitation of GMI1000 in tobacco leaves. All these results provide novel insights into understanding various biological functions of RsgA proteins and complex regulation on the T3SS in R. solanacearum.

Antitumor efficacy of interferon-γ-modified exosomal vaccine in prostate cancer.

BACKGROUND: Exosomes secreted by tumor cells can be regarded as carriers of tumor-associated antigens and have potential value in tumor immunotherapy. The aim of this study was to evaluate the antitumor efficacy of a novel exosomal vaccine (interferon-γ [IFN-γ]-modified exosomal vaccine) in prostate cancer. METHODS: Prostate cancer cell-derived exosomes were used to prepare the exosomal vaccine using our protein-anchoring technique. The immunogenicity and therapeutic efficacy of the exosomes was evaluated by measuring the effects of the exosomal vaccine on M1 macrophage differentiation, the ability of macrophages to engulf the exosomes, the production of antibodies against exosomes, and tumor angiogenesis and metastasis, and tumor growth. RESULTS: The IFN-γ fusion protein was efficiently anchored on the surface of prostate cancer cell-derived exosomes and retained its bioactivity. The IFN-γ-exosomal vaccine increased the number of M1 macrophages, enhanced the ability of M1 macrophages to engulf RM-1 cell-derived exosomes, and induced the production of specific antibodies against exosomes. The exosomal vaccine downregulated the expression of vascular endothelial growth factor receptor 2 and attenuated the effect of exosomes in promoting tumor metastasis. The proportions of CD4+ , CD8+ , and IFN-γ+ CD8+ T cells in the exosomal vaccine group were the highest among the four groups. Interestingly, the IFN-γ-exosomal vaccine decreased the percentage of Tregs and downregulated the expression of programed death-ligand 1 and indoleamine 2, 3-dioxygenase 1 in the tumor environment. The exosomal vaccine significantly inhibited tumor growth and prolonged the survival time of mice with prostate cancer. The exosomal and tumor cell vaccines had a good synergistic effect in promoting tumor immunity. CONCLUSIONS: The novel exosomal vaccine induced an immune response that cleared prostate cancer cell-derived exosomes, thereby eliminating the regulatory effect of the exosomes. This study may provide experimental evidence for the use of exosomes as a therapeutic tool or target in immunotherapy for human prostate cancer.