bluuebunny/crossref_metadata_2025 huggingface dataset:
Readability Evaluation of Books in Chinese as a Foreign Language Using the Machine Learning Algorithm
Authors: Ji Qiong
Published: September 2022

The arrival of the boom of teaching Chinese as a foreign language (TCFL) and personalized learning has led to a sharp increase in the demand for the Chinese language reading material. There are numerous reading materials available in Chinese for foreign students and learners to read and evaluate. The high-quality TCFL reading materials with reasonable arrangement can provide convenience for learners with different reading comprehension, interpretation abilities, and levels to master a language more quickly. Therefore, this study carries out an automatic readability assessment of books in Chinese as a foreign language. This paper comprehensively considers the factors affecting the difficulty of reading materials from the perspective of Chinese ontology based on the existing readability assessment research. Using natural language processing and a database management system to extract the features of books in Chinese as a foreign language, the text readability is evaluated with a statistical machine learning algorithm. The model is optimized by feature selection and sorting feature selection technology. The packaging feature selection technology is introduced to optimize the algorithm performance. The feature sets and each independent feature in the three dimensions of word meaning, part of speech, and discourse were optimized by the machine learning regression model based on certain evaluation indexes. Moreover, this work examined that the regression model is effective at identifying and recommending simpler textbooks for learning with difficult foreign language materials. For high-proficient learners, this approach significantly improves performance and measurement efficiency of reading books.
Convergence Spatial Measurement of Economic Growth Based on Big Data
Authors: Jin Jianglei , Yan Haowen , Zhao Qianqian
Published: September 2022

Economic growth has always been one of the hottest topics in economic research. Behind the rapid economic growth, the economic gap between regions is gradually widening, and the internal gap will have an impact on the overall coordination of economic growth. Research on the convergence of economic development and its causes has great strategic significance for narrowing the differences in raising economy among regions. In recent years, the impact of big data on economic analysis has become more and more obvious, and this fact has attracted the attention of the academic community. Big data are a new strategic resource and a tool for assessing economic trends. Adding big data technology to the research on the convergence raise of economics can predict the law of data changes, reduce data errors, optimize research results, and provide a more scientific basis for the coordinated development of regional economies. Based on big data theory and technology, this paper uses a spatial econometric model to empirically analyze the convergence of regional economic growth and its influencing factors. The experimental results show that the research on the convergence mechanism and spatial relationship of economic growth in the context of big data can improve the accuracy of the convergence analysis of economic growth to a certain extent. Through modeling analysis, the accuracy of economic convergence is improved by 4.1%. The utilization of big data in a trend of economic development makes the analysis results more reasonable and has greater reference value.
Human Placenta-Derived Mesenchymal Stem Cells Ameliorate Diabetic Neuropathy via Wnt Signaling Pathway
Authors: Pan Songsong , Hada Sushant S. , Liu Yang , Hu Chao , Zhou Mengdie , Zheng Shaoqiu , Xu Minjie , Shi Changsheng , Yin Shiwu , Xie Xiaoyun
Published: September 2022

Objectives. To investigate the effect of placenta-derived mesenchymal stem cells (PMSCs) on diabetic peripheral neuropathy and explore the role of Wnt signaling pathway. Method. Twenty-seven male db/db mice were randomly categorized into the control group, PMSC group, and PMSC treatment with Wnt inhibitor treatment group. Intervention was initiated in week 22. Thermal stimulation response was determined with a plantar analgesia tester. The mice were sacrificed on 7, 14, and 28 days. The morphology of sciatic nerves was observed by electron microscopy, and the expression of protein gene product (PGP) 9.5, S100β, and Ku80 was detected by immunofluorescence. Bax, β-catenin, and dishevelled1 (DVL1) were detected by western blot. Results. Thermal stimulation response was improved in the PMSC group on 14 and 28 days. Compared with the control group, PGP9.5 was increased in the PMSC group, accompanied by a significant increase in the expression of S100β. On the contrary, LGK974 inhibited the effect of PMSCs on thermal stimulation response and the expression of PGP9.5 and S100β. Both PGP9.5 and S100β were correlated with Ku80 in fluorescence colocalization. The myelin sheath of sciatic nerves in the PMSC group was uniform and dense compared with that in the control group. The effects of PMSCs promoting myelin repair were significantly inhibited in the PMSC+LGK974 group. Bax in the PMSC group expressed less than the control group. In contrast, the expressions of β-catenin and DVL1 were higher compared with that in the control group on the 14th and 28th days. The expression of DVL1 and β-catenin was lower in the PMSC+LGK974 group than in the PMSC group. Conclusions. PMSCs improved the symptoms of diabetic peripheral neuropathy, along with the improvement of nerve myelin lesions, promotion of nerve regeneration, and activation of Schwann cells, which might be related to the regulation of Wnt signaling pathway and inhibition of apoptosis.
Analysis of the Mechanism of Maslinic Acid on Papillary Thyroid Carcinoma Based on RNA-Seq Technology
Authors: Li Rong , Zhang Yanjiao , Xiang Runqing , Lin Aihe , Xia Zongxiao , Long Xiaomei , Guo Shuang , Fan Yuan , Chen Zukun
Published: September 2022

Objective. This study analyzed gene sequence changes in the thyroid papillary carcinoma (PTC) cell line TPC-1 treated with the natural compound maslinic acid (MA) through RNA-sequencing (RNA-seq) and identified the necessary genes to provide a basis for the study of the molecular mechanism of action of MA in PTC treatment. Methods. RNA-seq technology was used to detect genetic differences between the normal cell group (Nthy-ori 3-1) and the TPC-1 cell group (N vs T). Then, gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, Venn diagram analysis of shared genes, and protein–protein interaction (PPI) network analysis were used to analyze the therapeutic effect of the MA on TPC-1 cells. Real-time quantitative PCR (qRT-PCR) was used to verify six key genes. Results. GO and KEGG analyses showed that four crucial signaling pathways are related to TPC development: cytoplasmic molecule (cell adhesion molecules), neuroactive ligand–receptor interaction, tumor transcriptional disorder, and cytokine–cytokine interaction. The Venn diagram revealed 434 genes were shared between the MA vs T-group and 387 genes were shared between the MATH vs T and N vs T groups. PPI and ClueGO showed that NLRP3, SERPINE1, CD74, EDN1, HMOX1, and CXCL1 genes were significantly associated with PTC, while CXCL1, HMOX1, and other factors were mainly involved in the cytokine–cytokine interaction. The qRT-PCR results showed that the expression of NLRP3, EDN1, HMOX1, and CXCL1 genes was significantly upregulated in the TPC-1 group but significantly downregulated after MA treatment (

p
<
0.01

). SERPINE1 and CD74 genes were not expressed in TPC-1 cells, whereas they were significantly upregulated after MA treatment (

p
<
0.01

). Conclusions. This present study proves for the first time that MA can treat PTC, and the preliminary identification of key genes and rich signal transduction pathways provides potential biomarkers. It also provides potential biomarkers for the treatment of PTC with the natural compound MA and preliminarily discusses the therapeutic mechanism of action of MA against PTC, which is helpful for the further diagnosis and treatment of PTC patients.
3D Semantic VSLAM of Dynamic Environment Based on YOLACT
Authors: Wu Dongyan , Xie Bingbo , Tao Chongben
Published: September 2022

A Visual Simultaneous Localization and Mapping (VSLAM) method is proposed to construct a 3D environment map in the creation of dynamic noise information which leads to significant errors in camera pose estimation and a substantial number of noise points. For this problem, this paper proposed a method based on YOLACT, which combined optical flow and ViBe+ semantic map construction algorithm. First, our approach uses LK optical flow method to estimate the overall motion trajectory of adjacent frames. Then the trajectory data are employed to intercept the relative position of the current frame to the previous frame. Afterward, we combine ViBe+ algorithm to accurately detect and eliminate dynamic noise. Secondly, image semantic segmentation is performed based on YOLACT model. Image feature points are extracted from MAPLAB algorithm, pose estimation of the camera is performed and movement trajectory is recorded to complete a semantic map. Finally, through ablations study with common algorithms, the experimental results show that the proposed algorithm effectively avoids interference of dynamic noise information on VSLAM. Additionally, the constructed semantic map provides higher precision, fewer noise points, and pretty robustness.
The Roles of Tumor-Associated Macrophages in Prostate Cancer
Authors: Han Chenglin , Deng Yuxuan , Xu Wenchao , Liu Zhuo , Wang Tao , Wang Shaogang , Liu Jihong , Liu Xiaming
Published: September 2022

The morbidity of prostate cancer (PCa) is rising year by year, and it has become the primary cause of tumor-related mortality in males. It is widely accepted that macrophages account for 50% of the tumor mass in solid tumors and have emerged as a crucial participator in multiple stages of PCa, with the huge potential for further treatment. Oftentimes, tumor-associated macrophages (TAMs) in the tumor microenvironment (TME) behave like M2-like phenotypes that modulate malignant hallmarks of tumor lesions, ranging from tumorigenesis to metastasis. Several clinical studies indicated that mean TAM density was higher in human PCa cores versus benign prostatic hyperplasia (BPH), and increased biopsy TAM density potentially predicts worse clinicopathological characteristics as well. Therefore, TAM represents a promising target for therapeutic intervention either alone or in combination with other strategies to halt the “vicious cycle,” thus improving oncological outcomes. Herein, we mainly focus on the fundamental aspects of TAMs in prostate adenocarcinoma, while reviewing the mechanisms responsible for macrophage recruitment and polarization, which has clinical translational implications for the exploitation of potentially effective therapies against TAMs.
Statistical and SWAT Model-Based Performance Evaluation of RCMs in Modeling Streamflow and Sediment Yield at Upper Awash Sub-Basin, Ethiopia
Authors: Chelkeba Tumsa Bekan
Published: September 2022

The focus of this study was to evaluate the performance of the regional climate models with regard to simulating streamflow, sediment yield, precipitation, and temperatures. It is recognized that RCMs are not free of bias and uncertainty when simulating climate variables. The evaluation was about simulating annual climatology, annual cycles, and annual variability of climate variables by statistical tools and streamflow and sediment yield by SWAT model output. The study used observed and CORDEX Africa-44 meteorological data for RACMO22T, RCA4, CCLM4-8-17, and HIRHAM5 models using grid points. This analysis of the mean annual rainfall cycle in the summer season shows that all RCMs were underestimated. However, RACMO22T and RCA4 are better suited for simulating climate variables. The higher errors were associated with the simulations of maximum and minimum temperatures in the highest terrain area of the catchment. The statistical analysis with climatology indicates that all RCM was performed in much the same way, except for the seasonal perspective. In this case, RACMO22T was best able to simulate streamflow and sediment yield with PBIAS of 0.14, NSE of 0.91, R2 of 0.82, R2 of 0.72, NSE of 0.78, and PBIAS of −2.61%, respectively. RCA4 simulated streamflow better, but it underestimated the simulated sediment yield. The result proved that RACMO22T and RCA4 performed better in the upper floodplain area. The performance of the climate model varied with catchments, locations, and terrains. The output of this statistical and SWAT model shows that climate models do not accurately simulate hydro-climatological variables. Finally, this study showed that climate models were better at simulating the rainy season than the dry season. This integration of statistical tools and the SWAT model to analyze the RCM’s performance is a unique method to improve the quality of the output for its implementation in maintaining water balance and sediment load reduction.
Molecular Mechanisms of Gynostemma pentaphyllum in Prevention and Treatment of Non-Small-Cell Lung Cancer
Authors: Liang Renji , Wu Jinzheng , Lin Ronghua , Ran Liling , Shu Bo , Deng Hao
Published: September 2022

Objective. Lung cancer represents the leading cause of cancer death on a global scale. Gynostemma pentaphyllum (G. pentaphyllum), a traditional medicinal material with a high medicinal and health value, has recently been reported for its anticancer activity. However, the pharmacological mechanism of G. pentaphyllum in non-small-cell lung cancer (NSCLC) remains to be elucidated. Methods. The active ingredients of G. pentaphyllum were obtained from the TCMSP database and known therapeutic targets of NSCLC from the GeneCards and OMIM databases. Disease-drug common targets are subjected to protein-protein interaction (PPI), GO enrichment analysis, and KEGG pathway enrichment analysis. A molecular docking strategy was performed to verify the interaction between molecules. Results. We found a total of 24 compounds of G. pentaphyllum fulfilling OB ≥ 30% concomitant with DL ≥ 0.18 and corresponding 81 target genes in the TCMSP database, with 5062 NSCLC-related genes collected in the GeneCards and OMIM databases. The network consisting of the disease-target compound was obtained, including 8 active ingredients and 69 common targets. The PPI network with 65 nodes and 645 edges was visualized. After functional enrichment analysis, it was revealed that the therapeutic effects of G. pentaphyllum on NSCLC were achieved through response to ketone, gland development, and cellular response to xenobiotic stimulus. After molecular docking analysis, it was revealed that the two active ingredients of G. pentaphyllum, quercetin and rhamnazin, bound well and stably to their targets (MYC, ESR1, and HIF1A). Conclusion. Our study, based on network pharmacology, identifies active ingredients, targets, and pathways model mechanism of G. pentaphyllum when it is used to treat NSCLC.
A New View of Activating Mutations in Cancer
Authors: Nussinov Ruth , Tsai Chung-Jung , Jang Hyunbum
Published: September 2022

A vast effort has been invested in the identification of driver mutations of cancer. However, recent studies and observations call into question whether the activating mutations or the signal strength are the major determinant of tumor development. The data argue that signal strength determines cell fate, not the mutation that initiated it. In addition to activating mutations, factors that can impact signaling strength include (i) homeostatic mechanisms that can block or enhance the signal, (ii) the types and locations of additional mutations, and (iii) the expression levels of specific isoforms of genes and regulators of proteins in the pathway. Because signal levels are largely decided by chromatin structure, they vary across cell types, states, and time windows. A strong activating mutation can be restricted by low expression, whereas a weaker mutation can be strengthened by high expression. Strong signals can be associated with cell proliferation, but too strong a signal may result in oncogene-induced senescence. Beyond cancer, moderate signal strength in embryonic neural cells may be associated with neurodevelopmental disorders, and moderate signals in aging may be associated with neurodegenerative diseases, like Alzheimer's disease. The challenge for improving patient outcomes therefore lies in determining signaling thresholds and predicting signal strength.
Abstract 003: Treatment With
Ligilactobacillus
Improves Gut Permeability And Hypertension In Spontaneously Hypertensive Rats
Authors: Mukohda Masashi , Yano Takanori , Matsui Toshiyasu , Nakamura Sho , Miyamae Jiro , Mizuno Risuke , Ozaki Hiroshi
Published: September 2022

Emerging evidence provide that a microbial imbalance (dysbiosis) is linked to several diseases including cardiovascular diseases. One of the features in hypertension is increased gut permeability. Supporting this, plasma endotoxin level is increased in hypertension and our previous study indicated that the plasma level of exotoxin from
Streptococcus
was increased in spontaneously hypertensive rats (SHR). Thus, we hypothesized that improvement of gut permeability in hypertension might be expected to prevent development of hypertension. In this study, we performed a 16S metagenome analysis using a synthetic long read sequencing approach by LoopSeq in feces from 10- to 15-week old SHR and Wistar-Kyoto rat (WKY). The analysis revealed that two difference species of anti-inflammatory microbiome were strongly decreased in SHR compared with WKY (n=4). We focus on the decreased
Ligilactobacillus
sp. (LAB) in SHR and oral administration of LAB to SHR for 2 weeks. SHR (10-15-week-old) exhibited increased gut permeability in proximal colon (P<0.05, n=4), which was improved by treatment with LAB for 2 weeks (P<0.05, n=4). The expression level of tight junction protein Claudin4 was decreased in SHR proximal colon (SHR vs WKY, 1.0±0.1 vs 1.7±0.2-fold increase to SHR, P<0.05, n=8), which was significantly increased by LAB treatment (SHR vs SHR+LAB, 1.0±0.1 vs 2.8±0.5-fold increase to SHR, P<0.05, n=10). Proximal colon in SHR increased the expression level of TNF-α mRNA (1.0±0.2 vs 0.2±0.1, P<0.05, n=8), which was reversed by LAB (1.0±0.2 vs 0.3±0.1, P<0.05, n=10). The level of mucin 2 was lower in SHR proximal colon (1.0±0.2 vs 4.3±0.8, P<0.05, n=8), which was not reversed by LAB (1.0±0.1 vs 0.8±0.3, n=10). We observed that high blood pressure in SHR was attenuated by treatment with LAB for 2 weeks (194.0±5.9 vs 149.7±4.8 mmHg, P<0.05, n=10). LAB also improved decrease in acetylcholine-induced relaxation in SHR mesenteric artery (P<0.05, n=5-8). We conclude that treatment of SHR with the decreased anti-inflammatory microbiota, LAB improves gut permeability in proximal colon possibly through inhibition of inflammation, resulting in lowering blood pressure. These results suggest that anti-inflammatory microbiome might be a new therapeutic target for hypertension.
Abstract 005: Loss Of Resident Renal Dendritic Cells Prevents Salt-sensitive Hypertension And Pro-inflammatory Cytokine Production
Authors: Yoshigi Masaaki , Stuart Deborah , Koehler Kennedy , Srinivasan Harini , Eaton Douglas C , Ramkumar Nirupama , Wynne Brandi M
Published: September 2022

Hypertension (HTN) is associated with an increase in pro-inflammatory cytokines. Sentinel renal dendritic cells (rDCs) form an intricate network within the kidney cortex. DCs can activate immune cells and secrete pro-inflammatory cytokines. We hypothesize that loss of renal-specific DCs (rDCs) prevents salt-sensitive hypertension (SS HTN) and reduces intra-renal chemokine and cytokine production. To test this hypothesis, male mice lacking a functional CX
3
CR
1
chemokine receptor (CX
3
CR
1
-
EGFP+/+
) or wild-type (Wt) were used. CX
3
CR
1
is required for DC localization to the kidney; thus these mice have a renal-specific DC-depletion. Mice were subjected to L-NAME (0.5mg/mL, 2 wks) and then a wash-out (1-2wks), followed by HS (4%) or normal chow (NC) for the remainder of the study. Mice were implanted with telemetry devices and mean arterial pressure (MAP) assessed. Total kidney mRNA was isolated and pro-inflammatory chemokine/cytokine levels were determined using RT-PCR (n=4-6). Wt mice exhibited a progressive increase in MAP, with rDC-depleted mice having significantly lower MAP by week 3 (121±4
vs
. 114±3mmHg, *p<0.05). We observed a significant increase in renal mRNA levels of IL-27 (3.24 fold change **p<0.01) and IL-5 (3.76 fold change ****p<0.001) in Wt mice during SS HTN. Renal levels of IL-5 (1.69 fold change *p<0.05), IL-33 (0.72 fold change *p<0.05) and CXCL10 (0.52 fold change *p<0.05) were significantly reduced in rDC-depleted mice during SS HTN, as compared to Wt. When compared to NC, IL-10 levels were unchanged following SS HTN and no differences observed in rDC-depleted mice. Here, we show that rDC-depletion reduces the SS HTN response and that rDCs are necessary for the intra-renal increase of some pro-inflammatory chemokines and cytokines. These data suggest that sentinel innate immune cells within the kidney are crucial in the pathogenesis of SS HTN.
Abstract 008: Genetic Studies Identify A Novel Role Of
CCDC93
In Arterial Relaxation And Central Systolic Blood Pressure
Authors: Kumar Nitin , Yang Min-Lee , Sun Pengfei , Hunker Kristina , Zhang Yan , Ganesh Santhi K
Published: September 2022

Previous genome-wide association studies (GWAS) have identified and validated variants at more than 1000 loci with modest effects on population blood pressure (BP), explaining in aggregate ~6% of the trait variance and highlighting genetic differences in BP regulation in ethnically diverse populations. We performed an exome-wide association study (EWAS) in Han Chinese population (N=5,954) from Beijing and identified 4 EWAS single nucleotide polymorphism (SNP)-trait associations with 3 SNPs, including two novel associations (rs2165468-peripheral systolic BP and rs33975708-central systolic BP (cSBP). rs33975708 is a coding variant in the coiled-coil domain containing 93 (
CCDC93
) gene, c.535C>T, p.Arg179Cys (MAF=0.15%) and was associated with substantially increased cSBP (β=29.3 mmHg, P =1.23E-7) in Han Chinese population. CRISPR/Cas9 was used to introduce this variant in mice to investigate vascular molecular mechanisms. While this did not yield a viable mouse with the specific SNP, we did generate mice with an out of frame deletion making a gene knock-out.
Ccdc93
deletion (
Ccdc93
-/-
) in its homozygous form was embryonic lethal (<E10.5).
Ccdc93
+/-
heterozygous mice were viable and morphologically normal and displayed 1.31-fold lower aortic
Ccdc93
protein expression (P=0.0041, N=6). Systolic BP was higher in
Ccdc93
+/-
mice (110±8 mmHg vs 125±10 mmHg, P=0.016, N=5-7) and showed impaired acetylcholine (Ach)-induced relaxation (max Ach 86% vs 68%, P<0.03) and enhanced phenylephrine (PE)-induced contraction (max PE 123% vs 198% normalized to KCL response, P<0.03, N=5) in thoracic aorta
ex vivo
. Transcriptome analysis by RNA-seq of thoracic aorta identified significantly enriched biological processes in
Ccdc93
+/-
among the pathways altered in fatty acid metabolic process (adjusted P=2.9E-9) together with mitochondrial metabolism (P=1.3E-14). Plasma free fatty acid levels were higher in
Ccdc93
+/-
mice (96±7 μM vs 124±13 μM, P=0.0031, N=6) and aortic parkin protein expression, a marker of mitochondrial dysfunction, was 1.63-fold higher (P=0.028, N=6). In conclusion, our genetic and functional studies indicate a potential novel role of CCDC93 on blood pressure regulation through its effects on vascular mitochondrial function.
Abstract 026: Sex Specific Regulation Of The Sodium-chloride Cotransporter And The Salt Sensitivity Of Blood Pressure In Age-dependent Hypertension
Authors: Kim Kiyoung , Wainford Richard D
Published: September 2022

Objectives:
Blood pressure (BP) increases with age in a sex-dependent manner and a high salt diet (HSD) raises BP in salt sensitive individuals. The Na
+
-Cl
-
cotransporter (NCC) in renal cortex contributes to the fine-tuning of sodium reabsorption and BP regulation. We hypothesized that 1) increased NCC activity and expression contributes to sex- and age-dependent hypertension (HTN), and 2) age impairs HSD evoked suppression of NCC activity and expression with age to drives the development of salt sensitive hypertension (SSH).

Methods:
Three different age groups (3, 8, 16 month old) of male and female Sprague-Dawley (SD) rats were fed a normal salt diet (NSD; 0.6% NaCl) or HSD (4% NaCl) for 21 days. On day 21, basal MAP and NCC activity (peak natriuresis to IV hydrochlorothiazide (2mg/kg) infusion) were measured in vivo. The expression of NCC, with-no-lysine [K] kinases (WNK) 1, WNK4, STE20/SPS1-related proline-alanine-rich protein kinase (SPAK), oxidative stress responsive kinase 1 (OxSR1), and their phosphorylation status was assessed via immunoblotting (N=6/gp)

Results:
Male, but not female, rats develop age-dependent HTN with increased NCC activity and increased expression of NCC, WNK1, SPAK and phosphorylated SPAK/OxSR1 with age during NSD. A HSD evokes SSH with age in male, but not female, rats, and HSD evoked suppression of NCC activity and expression, WNK1, SPAK, OxSR1 and their phosphorylation is impaired in aged male rats.

Conclusion:
These findings suggest that 1) increased NCC activity and expression contributes to age-dependent HTN in male, but not female, SD rats, and 2) HSD evoked suppression of NCC is impaired with age driving SSH in male but not female SD rats.
Abstract 028: Toll-Like Receptor 4 Promotes Renal Immune Cell Infiltration And The Development Of Renal Damage During Dahl Salt-Sensitive Hypertension
Authors: Burns Emily C , Walton Samuel , Dasinger John H , Shaw Mary , Fehrenbach Daniel J , Geurts Aron M , Abais-Battad Justine M , Mattson David L
Published: September 2022

Previous work in our laboratory has implicated a significant role for the immune system in potentiating the hypertensive and kidney damage phenotypes which occur in the Dahl Salt-Sensitive (SS) rat. Toll-like receptors (TLRs), usually expressed on macrophages, play a key role in the innate immune system. A number of TLRs have been implicated in cardiovascular disease and chronic kidney disease with particular interest in TLR4, which specifically binds gram-negative lipopolysaccharide or endogenous molecules produced as a result of tissue injury. We previously performed a transcriptomic analysis which demonstrated TLR4 upregulation in renal medullary tissue of Dahl SS rats fed a high salt (4.0% NaCl) diet. The present study tests the hypothesis that TLR4 influences high salt diet-induced hypertension and renal damage in the Dahl SS rat. Using CRISPR/Cas9 technology, a mutation was generated in the TLR4 gene on the Dahl SS background, creating a 1-bp frameshift deletion resulting in a premature stop codon. SS
TLR4-/-
males and parental Dahl SS males (SS
TLR4+/+
), maintained on a 0.4% NaCl chow, underwent carotid artery telemetry implantation at 7 weeks of age. After recovery and baseline mean arterial pressure (MAP) recordings (day 0: 125.6 ± 0.5 vs 124.2 ± 2 mmHg), animals were switched to a 4.0% NaCl chow for 3 weeks. Overnight urine collections occurred once during the low salt period and weekly throughout the high salt challenge. Kidneys were flushed prior to euthanasia. Although SS
TLR4-/-
MAP was not different from SS
TLR4+/+
during the high salt challenge (day 21: 176.8 ± 14 mmHg, n=2 vs 172.3 ± 18 mmHg, n=4), high salt day 21 urinary albumin excretion was significantly reduced in SS
TLR4-/-
(272.7 ± 65 mg/day, n=5, p<0.01) compared to SS
TLR4+/+
(427.5 ± 67 mg/day, n=7), indicating reduced renal damage in TLR4 absence. Furthermore, flow cytometric analysis of isolated renal immune cells indicated a 33% reduction in CD45+ total leukocytes (5.0 ± 1.2 vs 3.4 ± 0.9 x10
6
cells) and a 31% reduction in CD11bc+ monocytes/macrophages (3.9 ± 0.9 vs 2.6 ± 0.7 x10
6
cells) in SS
TLR4-/-
(n=5) vs SS
TLR4+/+
(n=7) kidneys. Together, these data indicate that TLR4 may drive the immune response during the induction and progression of salt-sensitive hypertension and renal disease.
Abstract 029: Loss Of The Circadian Protein Per1 In The Dahl Salt-sensitive Rat Causes Exacerbation Of Diurnal Blood Pressure And Disruption Of Electrolyte Homeostasis
Authors: Zietara Adrian , Dissanayake Lashodya , Levchenko Vladislav , Gumz Michelle L , Staruschenko Alexander
Published: September 2022

Circadian rhythm is the 24-hr cycle experienced by most organisms which influences many physiological processes. Rhythms in physiological function are governed by a molecular circadian clock. Disruption of the circadian rhythm has been linked to deleterious cardiovascular outcomes, including the exacerbation of hypertension. The circadian clock is maintained by a group of transcription factors, including the protein Period 1 (encoded by PER1 gene), which regulate the expression of many different target genes in a tissue-specific manner. Previous studies in mice have shown that knockout of PER1 affects their ability to maintain proper sodium and blood pressure homeostasis. We hypothesized that the global knockout of PER1 on the Dahl salt-sensitive background (SS
Per1-/-
) will affect the development of salt-sensitive blood pressure. Following the high salt diet, SS
Per1-/-
rats experienced reduced weight gain (344 ± 7 vs 280 ± 9 g, p<0.001), excreted less sodium (17.3 ± 1.1 vs 12.1 ± 1.0 Na
+
/Cre, p=0.021), and exhibited reduced plasma potassium (3.6 ± 0.1 vs 3.0 ± 0.1 mM, p=0.005) and creatinine clearance (126.0 ± 23.8 vs 64.7 ± 5.9 mL/hr, p=0.008). Additionally, the SS
Per1-/-
group displayed increased renal tissue damage quantified by cortical fibrosis (0.9 ± 0.4 vs 6.2 ± 1.6 % area, p=0.011) and medullary protein casts (7.4 ± 0.9 vs 24.3 ± 4.0 % area, p=0.003). Examining the diurnal mean arterial pressure revealed no difference between SS and SS
Per1-/-
during their inactive day/light cycle (121.0 ± 1.8 vs 122.0 ± 1.9 mmHg, p=0.792), or their active night/dark cycle (127.2 ± 2.2 vs 127.6 ± 1.4 mmHg, p=0.883) when on a normal salt diet. However, after three weeks on a high salt 4% NaCl diet, SS
Per1-/-
animals experienced an exacerbated blood pressure during both the light (167.5 ± 3.7 vs 190.9 ± 7.2 mmHg, p=0.020) and dark (177.7 ± 3.4 vs 195.4 ± 4.5 mmHg, p=0.014) cycles. In the following analysis, we compared tissues, plasma, and electrolytes from SS
Per1-/-
and SS rats collected at 2 am and 2 pm. The conducted analysis revealed a number of pathways differently regulated during day and night cycles. In summary, our data indicate that global knockout of PER1 exacerbates salt-sensitive blood pressure development, disrupts electrolyte homeostasis, and aggravates renal damage.
Abstract 040: Prevalence Of Dual Indications For Antihypertensive Medications And Statins In The National Health And Nutrition Survey
Authors: GUPTA KASHVI , Byrd J B , Brook Robert D , Rubenfire Melvyn , Dorsch Michael P , Nallamothu Brahmajee K , Murthy Venkatesh L
Published: September 2022

Introduction:
Despite widespread indications for concomitant treatment of hypertension and hyperlipidemia, the proportion of the U. S population eligible for both remains unknown. Further, although fixed-dose combinations of these drugs are associated with better adherence, few are available.

Hypothesis:
We hypothesized a significant proportion of the population would be eligible for antihypertensive medication and a statin.

Methods:
Data from the National Health and Nutrition Examination Survey from 2011 to 2018 with survey weighting to represent the noninstitutionalized U.S. population ≥18 years were utilized. Prevalence of individuals with dual indications for antihypertensive and statin therapy was determined using the 2017 ACC/AHA guidelines for hypertension and the 2019 ACC/AHA guidelines for primary prevention of cardiovascular disease, respectively.

Results:
Sociodemographic characteristics are shown in Table 1. Among those ≥18 years, there were 88 million (33.4%) individuals with an indication for antihypertensive medication and 88.5 million (33.9%) with an indication for a statin. 64.6 million adults or 24.8% (95% CI: 23.5 to 26.2) of the population had dual indications for antihypertensive medication and a statin. Notably, 73.4% of those indicated for antihypertensive medication were also indicated a statin.

Conclusions:
Quarter of the U.S. adult population and nearly three-quarters of hypertensive patients are eligible for antihypertensive medication and a statin. Combining antihypertensive medications with a statin in fixed-dose combinations could reduce medication disutility and increase adherence to optimize lipid and blood pressure goals.
Abstract 045: Soluble Protein Oligomers Induce Endoplasmic Reticulum Stress In Endothelial Cells And Lead To Decreased Calcium Levels In The Endoplasmic Reticulum.
Authors: Waigi Emily W , Pernomian Laena , Webb R. Clinton , G. McCarthy Cameron , F. Wenceslau Camilla
Published: September 2022

Soluble protein oligomers (SPOs) have powerful vascular effects suggesting a link between Alzheimer’s disease (AD) and cardiovascular diseases (CVD). SPOs accumulation induces endoplasmic reticulum (ER) stress hence cell death in several CVDs. Our previous data showed that in mesenteric resistance arteries (MRA), SPOs induce ER stress leading to pathologically exacerbated endothelium-dependent vasodilation. We therefore hypothesized that the SPOs will cause ER calcium dysregulation in endothelial cells isolated from MRA. To confirm the presence of ER stress and SPO subcellular localization, endothelial cells were treated with SPOs (0.1 υM) in the presence and absence of the ER stress inhibitor 4-PBA (2 mM) and vehicle (F-12 media). Immunostaining with KDEL antibody specific for ER resident chaperones Grp78/94 was performed for 19 hr prior to visualization. Endothelial cells treated with SPOs showed increased anti-KDEL fluorescence (AU: Vehicle: 4.74 ± 0.3 vs. SPO: 15.74 ± 1.7; n= 3; p= 0.018, one-way ANOVA). This is consistent with the increased Grp78/94 protein concentration during ER stress. Interestingly, 4-PBA treated cells reduced anti-KDEL fluorescence to the same level as the control (AU: Vehicle: 4.74 ± 0.3 vs. 4-PBA: 8.74 ± 2.9; n= 3; p= 0.39; one-way ANOVA) suggesting that SPOs bind to the ER-anchored proteins during ER stress. Additionally, changes in Ca
2+
concentrations have been implicated as a mediator between ER stress and SPOs. Calcium levels in endothelial cells was investigated by transfecting a plasmid expressing a Ca
2+
indicator pCMV G-CEPIA1
er
for 72 hr
.
Cells were treated with vehicle or SPOs in the presence or absence of 4-PBA (45 min) prior to fluorescence imaging. Cells treated with SPOs+ACh had a decreased fluorescence in the ER from the basal Ca
2+
levels (%Δ Ff-F0/F0; SPO: 53% reduction vs. vehicle), but treatment with 4-PBA+ACh did not improve this response (%Δ F
f
-F
0
/F
0
; SPO+4-PBA: 48.18% reduction vs. vehicle n=2). This reveals that SPOs induce ER stress and depletion of ER calcium. However, ER stress inhibitor did not improve Ca
2+
levels. These data suggest that ER stress, is the mechanism by which SPO’s exacerbate vasodilation in the mesenteric resistance artery endothelial cells regardless of changes in Ca
2+
levels.
Abstract 049: Abrogation Of Transglutaminase 2 In Vascular Smooth Muscle Cells Attenuates Doca/salt Induced Hypertension In Male Mice
Authors: Aroor Annayya , Manrique Camila , Naz Huma , Hulse Jack , Jia Guanghong , Whaley-Connell Adam , Lastra Gonzalez Guido
Published: September 2022

Background:
Transglutaminase 2 (TG2) is an enzyme involved in the pathogenesis of vascular remodeling in conditions of chronic over-nutrition and activation of the renin angiotensin aldosterone system (RAAS). Furthermore, it has been shown that pharmacologic inhibition of TG2 significantly reduces blood pressure in male rodents. However, the cell-specific role of TG2 activation in vascular smooth muscle cells (VSMC) in mediating the hypertensive response to RAAS overactivation has not been explored.

Methods:
We generated a novel inducible model (tamoxifen dose: 50 mg/kg/d oral for 6 weeks) of TG2 deletion in VSMC (VSMCTG2KO) by sequentially crossing Tgm2t/t floxed mice (loxP sites flanking exons 6-8 of the TG2, C polypeptide [Tgm2] gene) with male Myh11-CreERT2 positive mice. Effectiveness of the TG2 deletion was evaluated by immunohistochemistry and TG2 activity was determined by incorporation of Alexa Fluor488 Cadaverine.A cohort of fifteen-month-old wild-type and VSMC TG2KO male mice were implanted with deoxycorticosterone (DOCA; 50 mg, 21-day releasing pellets) and 1 % salt added to the drinking water. Systolic blood pressure was assessed by tail-cuff method prior to sacrifice. Ex vivo aortic stiffness was determined by atomic force microscopy. Student t-test or two-way ANOVA were used for statistical analysis as appropriate. The results were considered significant when p<0.05. Three to six mice were included per treatment group.

Results:
administration of DOCA/salt induced increases in systolic blood pressure, augmented expression of TG2 in aorta, as well as increased aortic stiffness in wild-type mice (10.47±1.27kPA DOCA/salt vs. 3.91
+
1.01 kPa placebo). The VSMCTG2KO model exhibited 77% downregulation of TG2 expression in VSMC. VSMC TG2 KO suppressed DOCA/salt induced TG2 activity (29.13
+
2.22 arbitrary units (AU) in DOCA/salt vs 14.10
+
2.90 AU in KO cohort). This was accompanied by a significant improvement in DOCA/ salt-induced hypertension (SBP 142 ± 5.4 mmHg in DOCA/salt treated mice vs. 111.8 ± 6.2 mmHg in VSMCTG2KO).

Conclusions:
Our data demonstrate for the first-time the key role of TG2 activation in VSMC, in the regulation of hypertensive responses in in a male rodent model of RAAS activation.
Abstract 055: Lipid Droplet Accumulating Microglia Involvement In Non-Alcoholic Fatty Liver Disease During Obesity
Authors: Kim Han Rae , Young Colin N
Published: September 2022

Non-alcoholic fatty liver disease (NAFLD), characterized by excess fat accumulation in the liver (i.e. steatosis), is directly associated with obesity and a risk factor for hypertension development. In addition to peripheral mechanisms, accumulating evidence points to alterations in the central nervous system as a NAFLD contributor. In this context, we recently demonstrated that selective inhibition of microglia, the resident immune cells of the brain, specifically in the subfornical organ (SFO), attenuated obesity-related hepatic steatosis by ~50%. However, underlying SFO microglia alterations that may contribute to NAFLD remain unknown. The SFO is a circumventricular nucleus situated outside of the blood-brain-barrier that is chronically exposed to circulating factors such as lipids. Interestingly, lipid droplet-accumulating microglia (LDAM) have been implicated in conditions such as Alzheimer's disease and aging. Based on this, we hypothesized that SFO LDAM may be a unique contributor to hepatic steatosis during obesity. To investigate this, 6 wk old C57Bl/6J male mice were fed a high fat diet (HFD, 60% kCal fat) or normal chow for 11 wks (Body weight: 33±1 vs 47±2 g, normal chow vs HFD, p<0.05, n=4-5/group). Immunohistochemistry for ionized calcium-binding adapter molecule 1 (Iba1) as a microglia indicator demonstrated an increase in SFO microglia of HFD fed mice (330±6 vs 393±18, normal chow vs HFD, p<0.05). Furthermore, combined immunohistochemistry for Iba1 and Perilipin-2 (Plin2) as a lipid droplet marker revealed an approximate doubling of SFO microglia with lipid droplet inclusion (Plin2+ microglia #: 80±11 vs 191±21; Plin2+ microglia %total: 23±3.2 vs 49±4.1, normal chow vs HFD, p<0.05). This increase in SFO LDAM was additionally confirmed with comprehensive 3-dimensional analysis of up to 400 microglia per animal using BODIPY, a dye that labels neutral lipids (e.g. HFD: 1.6±0.2 BODIPY+ microglia fold normal chow, p<0.05). Collectively, these findings indicate marked SFO microglia activation and microglia lipid accumulation during obesity and point to the possibility for SFO LDAM in the pathogenesis of NAFLD and associated hypertension development.
Abstract 056: Transcriptomic Changes In Individual Cell Types Of The Arcuate Nucleus In The Mouse Deoxycorticosterone-acetate (doca)-salt Model Of Hypertension
Authors: Wagner Valerie , Deng Guorui , Claflin Kristin , Cui Huxing , Nakagawa Pablo , Sigmund Curt D , MORSELLI LISA , Grobe Justin L , Kwitek Anne E
Published: September 2022

Low-renin hypertension is common among the elderly, subjects of black African ancestry, those with heart or renal failure, and women with preeclampsia. Increased understanding of the causes and consequences of low-renin hypertension is necessary for the development of new efficacious and personalized therapeutics. One commonly employed preclinical model is “DOCA-salt” (DOCA), which causes robust hypertension, proteinuria, and organ damage. The arcuate nucleus of the hypothalamus (ARC) has been implicated in control of metabolic rate and blood pressure responses to DOCA. To characterize the DOCA related changes within the ARC, we used single nucleus RNA sequencing (snRNA-Seq) to individually interrogate the transcriptomes of numerous mouse ARC cell types. Male C57BL/6J mice 7 weeks-of-age underwent sham surgery or subcutaneous implantation of a 50mg DOCA pellet. All mice had water; DOCA mice were also provided 0.15 M NaCl drink ad libitum. Body composition was assessed by nuclear magnetic resonance. After 3 weeks, tissues were weighed and blood chemistries assessed. Features of DOCA treatment were consistent with previous studies, such as suppressed growth trajectory (sham n=7, DOCA n=8; -0.8±0.4g/3wk; p<0.01). Nuclei were isolated from ARC punches for snRNA-seq. Thirty-two unique cell type clusters were identified from snRNA-Seq, and DOCA differential gene expression (DEG) was determined using MAST. Two clusters were classified as resting (Cluster 23: DEGS up=11, down=35) and activated (Cluster 25: DEGS up=45, down=169) microglia subtypes. Analysis of proportions suggests there are fewer activated microglia in the DOCA ARC (dataset Sham=25%, DOCA = 75%; Cluster 25 Sham=37%, DOCA=63%; χ2 p = 0.0056), and DOCA may negatively impact activated microglia functions through decreased calcium signaling and netrin receptor activity. Sub-clustering of neuropeptide clusters resulted in the identification of 3 Agouti-related peptide sub-clusters. DOCA DEG enrichment terms indicated decreased neurotransmitter uptake, altered GABA synapse functions, and changed hormone secretion in the sub-clusters, respectively. These data provide novel insights into cell-specific molecular changes occurring in the ARC due to DOCA-salt hypertension.
Abstract 063: Soluble (Pro)renin Receptor Acts Via Direct Activation Of Angiotensin Type 1 Receptor To Enhance Renal Aquaporin-2 Expression And Urine Concentrating Capability
Authors: Wang Fei , Lambert Jacob , Zheng Huaqing , Fu Ziwei , Zou Chang-Jiang , Yang Tianxin
Published: September 2022

Abundant evidence has established an essential role of collecting duct (pro)renin receptor (PRR) in regulation of aquaporin-2 (AQP-2) expression and urine concentrating capability. Moreover, such AQP2-regulating and antidiuretic action of PRR is mediated by its cleavage product, the soluble PRR (sPRR). However, the mechanism of action of sPRR in this setting remains incompletely understood. In light of our recent report of direct interaction of sPRR and angiotensin type 1 receptor (AT1R) in the vascular endothelial cells, we examined the hypothesis that sPRR signals via AT1R in the kidney. In primary rat inner medullary collecting duct cells, exposure to a recombinant sPRR (termed as sPRR-His) at 10 nM for 24 h induced 4 fold increase in AQP2 protein abundance, which was blunted by `~50% by pretreatment with losartan. In C57/BL6 mice, administration of the V2R antagonist (OPC at 30 mg/kg/d via gavage for 3 days) resulted in symptoms of nephrogenic diabetes insipidus (NDI), including polydipsia, polyuria, and hypoosmotic urine, all of which were improved by subcutaneous infusion of sPRR-His at 30 μg/kg/d via minipump (urine volume [ml]: 5.3 ± 0.2 vs. 3.3 ± 0.3, n =5, p < 0.01; urine osmolality [Osm/kg. H2O]: 428.6 ± 29.2 vs. 704.6 ± 24.2, n =5, p < 0.01). Administration of losartan (5 mg/kg/d) almost completely reversed the therapeutic effect of sPRR-His (urine volume [ml]: 4.7 ± 0.2; urine osmolality [Osm/kg. H2O]: 486 ± 16.1). Immunoblotting demonstrated that sPRR-His treatment restored OPC-induced reduction of AQP2 protein abundance, which was reversed by losartan. Together, sPRR interacted with AT1R to control renal AQP2 expression and urine concentrating capability.
Abstract 066: Renal-derived Human Sprr Induces Hypernatremia And Water Retention In Male Mice But Not Female Mice
Authors: Arthur Gertrude , Poupeau Audrey , Taylor Alexis , Nichols Kellea , Leachman Jacqueline , Loria Analia S , Osborn Jeffrey , Yiannikouris Frederique B
Published: September 2022

The soluble prorenin receptor (sPRR) plays an important role in fluid and electrolyte balance. In rodent models, sPRR contributes to aquaporin2(AQP2)-dependent antidiuretic action. However, there is a gap of knowledge concerning the functional role of locally produced human sPRR from the kidney. Hence, we evaluated the role of renal-derived human sPRR in fluid and electrolyte homeostasis.Human sPRR-Myc-tag transgenic mice were bred with mice expressing Hoxb7/Cre to selectively express human sPRR in the collecting duct (RHsPRR). RHsPRR and control (CTL) male and female mice were fed a standard diet for 10 months (n=8-11/group). Body weight was examined weekly, body composition monthly and water balance at study endpoint. Western blot analysis depicted the presence of human sPRR-Myc-tag (28 KDa) in the cortex and medulla of RHsPRR male and female mice validating the humanized mouse model.Renal-derived human sPRR did not change body weight in male or female mice (Male: CTL: 34±1, RHsPRR: 33±1 g; Female: CTL: 28±1, RHsPRR: 30±1 g) nor kidney function assessed by GFR (Male: CTL: 817±83, RHsPRR: 1088±163 μl/min/100gBW; Female: CTL: 1057±75, RHsPRR: 875±89 μl/min/100gBW). In male mice, renal-derived human sPRR significantly elevated plasma sodium (Male: CTL: 115±4, RHsPRR: 127±3 mmol/L; P<0.05) and ENaC gene expression (M: 1.7±0.7 and 3.7±0.9 2
-ΔΔCT
, P<0.05). Urine flow rate was decreased (Male: CTL: 1.0±0.2, RHsPRR: 0.6±0.2 ml; P<0.05) and AQP2 protein expression was increased in male RHsPRR mice compared to CTL (M: 0.13±0.05 and 0.44±0.12 AU, P<0.05), suggesting that renal-derived human sPRR elicits hypernatremia by stimulating sodium reabsorption via ENaC-dependent mechanism and water reabsorption by AQP2 expression.In female mice, renal-derived human sPRR did not change plasma sodium (Female: CTL: 120±7, RHsPRR: 118±5 mmol/L), ENaC gene expression (F: 1.2±0.2 and 1.4±0.1 2
-ΔΔCT
), urine flow rate (Female: CTL: 0.4±0.1, RHsPRR: 0.5±0.1 ml) or AQP2 protein expression (F: 0.20±0.03 and 0.20±0.06 AU, P<0.05).Our data suggest that renal cortical and medullary human sPRR regulates water and electrolyte balance in a sex specific-manner. Translationally, human sPRR could be a promising therapeutic target to treat hypernatremia in men.