bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanism of a2V-ATPase in Sterile Inflammation (91.15)
Authors: Kwong Christina , Gilman-Sachs Alice , Beaman Kenneth 
Published: January 2023

Abstract
Key to our understanding the immune response during cell injury is the recognition of the unique mechanism used by the body to detect and respond to cell destruction. We show data that a vacuolar ATPase (a2V) is responsible for detecting aseptic cell injury and directs the activities which can lead to both tissue repair and disease pathogenesis. Rapid clearance of cellular debris is a crucial event to prevent host proteins from being recognized by immune cells to prevent unnecessary inflammation. a2V is a key protein that is up-regulated during sterile inflammation. After membrane receptors detect proximal cell death, a2V migrates to the plasma membrane and the N-terminus (a2NTD) is cleaved and secreted into the extracellular space. a2NTD stimulates primary human monocytes to secrete IL-1β. a2NTD also stimulates THP-1 cells in a concentration-dependent manner to up-regulate pro-inflammatory genes including IL-8, MCP-1 and MMP-9. We show a2NTD binding to monocytes cell surface that is reversible in a competition binding assay via flow cytometry and fluorescent microscopy. These data show involvement of CD14 with a2NTD binding and possible interaction between a2NTD's receptor and the inflammasome. We describe a new pathway by which monocytes respond to tissue destruction. Secretion of a2NTD represents a vital pathway utilized by tissues to initiate immune responses after cell injury.
Mechanisms of social disruption stress induced glucocorticoid resistance (91.16)
Authors: Stiner LaTonia , Padgett David , Sheridan John 
Published: January 2023

Abstract
Social disruption (SDR) stress increases splenic CD11b+ myeloid cells, which produce high levels of proinflammatory cytokines upon Toll-like receptor ligation and show impaired nuclear translocation of the GC receptor (GCr) after corticosterone (cort) stimulation. To uncover the mechanism of GC resistance, p38 MAPK (p38) expression and phosphorylation were investigated along with phosphorylation of the GCr. To examine the mechanism(s) of enhanced proinflammatory cytokine responses, CD11b+ splenocytes from control (HCC) and SDR mice were stimulated with lipopolysaccharide in the presence of cort. SDR and HCC CD11b+ cells show similar levels of unphosphorylated p38 protein. CD11b+ cells from SDR mice exhibited an increase in phosphorylation of p38 and of the GCr. Since phosphorylation of p38 has been linked to GCr phosphorylation in humans, it is likely that a same phenomenon occurs here, though it must be confirmed. Since, phosphorylation of the GCr reduces steroid-induced cytokine repression; this may explain the enhanced cytokine expression. These data show that phosphorylated forms of p38 and GCr play a role in GC resistance.
Support: NIH grants NIMH/RO1MH046801-16 and NIDCR/T32DE014320-6
Itk acts as a negative regulator in mast cell activation (91.19)
Authors: Evans John W , Falk Markus , Schattgen Stefan A , Kosaka Yoko , Berg Leslie J 
Published: January 2023

Abstract
Mast cells are key effectors of allergic and innate immune responses and have been implicated as 'immuno-modulators' in adaptive immunity. Mast cell activation through the immuno-receptor, FceR1, propagates a complex signaling cascade which culminates in a broad range of effector functions. Central to this signal transduction are various protein tyrosine kinase families including members of the Tec family.
The Tec family kinases, Itk and Btk, have been extensively studied in T cells and B cells, respectively, and have been shown to play positive signaling roles. Deficiency in either of these two kinases results in diminished signal transduction downstream of their respective immuno-receptors leading to defects in development as well as effector function.
In contrast to T cells and B cells, mast cells express both Itk and Btk at similar levels. Btk-deficient mast cells exhibit a decreased effector function consisting of reduced degranulation and cytokine production. Furthermore, this phenotype is accompanied by diminished signal transduction downstream of FceR1, as shown by decreased Ca2+ flux and PLCγ phosphorylation.
Here we show that Itk-deficient mast cells display almost the opposite phenotype. These mast cells show enhanced levels of Th2-associated cytokines as well as some pro-inflammatory factors. We show that signal transduction events downstream of FceR1, including Ca2+ flux and PLCγ phosphorylation, are enhanced in the absence of Itk. Additionally, we show that Itk interacts with and phosphorylates the phosphatase, SHIP-1, a negative regulator in mast cells. Finally, we show that Itk co-localizes with a negative signaling complex in wild-type mast cells. Taken together, these data indicate that Itk can impact mast cell activation by dampening the Ag-mediated response. This work suggests a novel role for Itk as a negative regulator in mast cell signaling.
Modulation of the Lipoxygenase Pathway in Bone-Marrow-derived Dendritic Cells by Peroxisome Proliferator Activated Receptor γ. (91.21)
Authors: Hontecillas Raquel , Mysiak Sara , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Enzymes of the lipoxygenase (LOX) family generate lipid mediators that activate the nuclear receptor peroxisome proliferator activated receptor γ (PPAR γ) and regulate immune and inflammatory reactions. GMCSF upregulates the 5-LOX pathway whereas IL-4 suppresses 5-LOX and upregulates 15-LOX expression. The objective of this study was to evaluate whether, in addition of being a molecular target of the lipid mediators generated from arachidonate oxidation by LOX enzymes, PPAR γ regulates the expression of both 5- and 15-LOX. We show that during differentiation of BMDC in GMCSF and IL-4, the loss of PPAR γ resulted in upregulation of 15-LOX. Phenotypically, PPAR γ deficient BMDC differentiated in GMCSF and IL-4 expressed more CD40 and less CD36 than PPARγ-expressing cells. Also, PPAR γ-defficient cells differentiated in the presence of GMCSF alone had more 5-LOX mRNA than PPAR γ-expressing cells, which was paralleled by upregulation of PPAR δ. The loss of PPAR γ did not change the expression of 5-LOX activating protein. LPS stimulation of mature BMDC induced secretion of LTB4, a product of the 5-LOX pathway, in cells differentiated in GMCSF although the loss of PPAR γ did not have any effect on the amount of this lipid mediator released. Our results suggest that a negative feed-back loop is initiated following activation of PPAR γ that regulates further generation of more endogenous ligands.
Validation and application of a multiplex immunoassay for the detection of PGE2 induced inflammatory mediators of human decidual macrophages (92.15)
Authors: Casper John M , Senn Tennille , Cox Jennifer , Black Eric , Aronoff David M. , Mullenix Michael M. 
Published: January 2023

Abstract
A multiplex bead immunoassay was developed for the simultaneous quantitation of 8 inflammatory mediators including 6 cytokines (IL-1 β, IL-4, IL-6, IL-8, TNFα and IFNγ) and 2 eicosanoids (PGE2, and TXB2). The sensitivities of the cytokine assays are less than 5 pg/mL with the exception of IL-4 (19.9 pg/mL). The sensitivity of the PGE2 and TXB2 assays at 80% B/Bo are 64.2 and 65.9 pg/mL for PGE2 and TXB2 respectively. The antimicrobial actions of human decidual macrophages (DMs) isolated from the upper reproductive tract were characterized in the assay. Decidual macrophages are innate immune cells known to defend the upper reproductive tract from infection. PGE2 is a potent regulator of innate immunity in the reproductive tract during pregnancy and/or in response to infection. CD14+ DMs were incubated with bacteria, peptidoglycan or LPS and analyzed for the expression of the inflammatory mediators detected in the MultiBead(tm) multiplex immunoassay. The DMs produced PGE2 in response to bacteria, peptidoglycan or LPS with increased production of IL-8, TNFα, IL-6 and IL-1 β. The addition of 1 μM PGE2 decreased expression of TNFα and IL-1β and increased the expression of IL-6. The DMs produced PGE2 in response to inflammatory stimuli and modulated the expression of cytokines and eicosanoids in vitro. We conclude that PGE2 is involved in immunomodulation that may be relevant when PGE2 levels are increased during pregnancy and infection.
Bone marrow Ly6Chi Monocytes are recruited to injured kidney and differentiate into Ly6Clo profibrotic macrophages (92.7)
Authors: CASTANO ANA P , Lin Shuei Liong , Duffield Jeremy  S 
Published: January 2023

Abstract
Macrophages (M(s) play decisive roles in different models of organ fibrosis. Prevailing concepts support circulating monocyte heterogeneity, subpopulation discrete recruitment into injured tissues, differentiation into M(s, and possibly subpopulation discrete roles in injury and repair. Here we show discrete subpopulations of M(s in fibrotic kidney induced by unilateral ureteral obstruction (UUO) and their discrete functional roles in renal fibrosis. Fluorescence-activated cell sorting (FACS) of purified UUO kidney leukocytes reveals 3 populations of M(s separated by Ly6C: high, intermediate, and low. FACS of peripheral blood mononuclear cells (PBMC) showed a marked increase in Ly6chi-Mo in response to kidney injury. Depletion of M(s in CD11b-DTR mice during UUO fibrosis inhibits the progression of fibrosis by an indirect mechanism. However, Ly6clo kidney M(s were selectively depleted indicating their predominant role in fibrogenesis. To determine the origins of kidney M(s, we lineage-traced 3 different populations of monocytes (Mo) by adoptive transfer of CD45.1+ bone marrow (BM)-Ly6chi Mo or peripheral blood (PB) Ly6chi Mo or PB-Ly6clo-Mo into CD45.2 mice with UUO. Ly6Chi BM-Mo were avidly recruited to injured kidney, and differentiated into Ly6Chi, Ly6Cint, and Ly6Clo kidney M(s. PB-Mo also differentiated into 3 populations but 20-fold lower recruitment was observed. We characterized kidney-M(-subpopulation transcript levels by branched-chain-DNA-technology which demonstrated discrete differences in expression of genes including TNF(, IL1(, MIP2, MIP1(. We conclude that Ly6Chi BM-Mo are recruited to kidney by mobilization from BM, differentiate into 3 functionally discrete kidney M(s one of which is profibrotic, and resident M(s play a non-fibrogenic role in kidney injury.
CD34 is required for the infiltration of inflammatory cells into the colon in a mouse model of ulcerative colitis (93.2)
Authors: Maltby Steven J , Wohlfarth Carolin , McNagny Kelly , Hughes Michael R 
Published: January 2023

Abstract
The infiltration of eosinophils into gut tissue and release of granular mediators appears to play a critical role in the pathogenesis of inflammatory bowel diseases (IBD) such as ulcerative colitis. Using an allergic asthma model, we recently demonstrated that eosinophil migration into the lung requires surface expression of the sialomucin CD34 on mast cells and eosinophils. Based in part on these findings, we have investigated a similar role for CD34 in the migration of eosinophils and other inflammatory cells into the colon, and the effects of CD34 deletion on IBD development using a dextran sodium sulfate (DSS)-induced model of ulcerative colitis. We monitored weight loss, colon length, rectal bleeding and diarrhea and our findings demonstrate decreased disease severity in DSS-treated Cd34-/- mice. Post-mortem histology of DSS-treated mice also showed reduced inflammation of Cd34-/- colon compared to wild type controls. Interestingly, the percentage of peripheral blood eosinophils was reduced significantly more in wild type than Cd34-/- mice at the disease endpoint - corroborating a defect in the exit of eosinophils from circulation into the gut. These findings support the conclusion that CD34 plays a role in eosinophil migration into gut tissues and suggest that CD34 may serve as a viable target for pharmacological interventions across a range of inflammatory conditions.
Differential gene expression profiles in the spontaneously hypertensive rats administrated with anti-hypertensive drugs (93.20)
Authors: Park Min , Lee Kee-Mo , Kim Eun-Mi , Park Yong-Wook , Kee Seung-Jung , Kang Hyung-Sik 
Published: January 2023

Abstract
To identify and characterize the genes involved in the functional regulation of hypertension and arteriosclerosis-specific genes, changes in gene expression profiles were examined by DNA microarray from the spontaneously hypertensive rats (SHR) and normal the Wistar-Kyoto (WKY) rats after administration with anti-hypertension drugs, including angiotensin I-converting enzyme (ACE) inhibitor, enalapril and the calcium channel blocker, nifedifine. Among the genes that were distinct and differentially expressed in brain, heart and liver of SHR rats administrated with the drugs, 113 were specifically up-regulated, and 102 were down-regulated genes compared to those of WKY or SHR rats. The differential gene expression profiles in DNA microarray were corroborated by semi-quantitative RT-PCR. Of these genes, expression of TNFSF4, 6, 10, IL-6 receptor, IL-18 receptor accessory protein and CD24 genes was extensively exhibited in a hypertension-specific manner, and patterns for hypertension-associated gene expression was regulated by changes of their expression in angiotensin ¥± or H2O2-treated vascular smooth muscle cells.
These results will provide a fundamental basis to delineate the novel mechanisms, and a thorough investigation of the relationship between their expression and function will provide a better understanding in the development of new therapeutic drugs against cardiovascular diseases.
This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF-2006-005-J03003)
NFkB and IkB expression are modulated by sex steroids during alcohol liver inflammation (93.24)
Authors: Lee Eric G , Mickle Bethany , Chastain Lerin , Gallucci Randle 
Published: January 2023

Abstract
Women are more susceptible to alcoholic liver disease (ALD) than men. The release of cytokines from Kupffer cells during inflammation is associated with the pathogenesis of ALD, and chronically ethanol fed female rats express cytokine mediators differently than male counterparts. Differences in inflammatory gene expression in females may have a role in gender related ALD pathology. This study examines the effects of sex steroids on the NFkB and IL-6 pathways in chronically ethanol fed rats. Ovarectomized, female, Wistar rats implanted with osmotic pumps releasing estrogen or testosterone were fed liquid diets with and without ethanol for two and four weeks. Estrogen receiving animals had lower mRNA expression of IL-6Rα, SOCS-3, PIAS-3, C/EBPβ, IkBα, and IkBβ at 2 weeks with little difference observed at 4 weeks. Ethanol consumption induced lower mRNA expression of IL-6Rα, PIAS-3, IkBα, and IkBβ in both testosterone and estrogen receiving animals with the lowest levels consistently observed in the estrogen group. Nuclear NFkB was greatest in the estrogen groups and lowest in the ethanol consuming testosterone group. IkBα protein expression was greatest in the estrogen receiving groups. The combined effects of estrogen and ethanol resulted in the lowest mRNA expression and highest NFkB and IkBα protein expression. Our data suggest that testosterone and estrogen distinctively affect NFkB, IkBα, and IL-6R expression and function and the effect of testosterone may be protective in ALD.
Funded by NIH/NIAAA grant R03AA015198
Presence of the functional Caspase-12 allele in Indian subpopulations (93.25)
Authors: Hermel Evan , Klapstein Kevin D , Yavari Mehdy 
Published: January 2023

Abstract
In rodents, caspase-12 (casp12) is a negative regulator of interleukin-1 production. Most humans lack a functional CASP12 gene, with a non-functional variant (CASP12p1), found in 100% of the Caucasian and East Asian population, and in approximately 80% of people of African descent. However, 20% of sub-Saharan Africans carry an intact allele of CASP12, which produces a full-length non-catalytic proenzyme and is associated with an increased risk of sepsis. Using PCR-based single nucleotide polymorphism analysis and DNA sequencing, we examined CASP12 allele distribution in individuals from Central and Southern Asia and found that CASP12 was significantly present in members of the Dravidian language group (the predominant ethnic and language group of southern India), particularly persons from Tamil Nadu. The CASP12 allele also occurs at a higher frequency in individuals with the Australoid morphotype that predominates in southern India, as compared to those with the Caucasoid morphotype prevalent in central and northern India. The data suggest that there is a north-to-south gradient of increasing prevalence for the CASP12 allele. As the CASP12p1 allele has been rigorously selected for in populations outside of Africa and southern Asia, we hypothesize that a pathogen that exploits the inflammatory immune response in Africa and southern India may be exerting positive selective pressure on CASP12.
The role of inflammation in diet-induced obesity (93.4)
Authors: Alexander Lindsey A , Slotterbeck Brandon D , Najjar Sonia M , McInerney Marcia F 
Published: January 2023

Abstract
Metabolic syndrome, affecting up to 25% of Americans, includes impaired glucose tolerance, central obesity, dyslipidemia, insulin resistance, and type 2 diabetes. Secondary complications include atherosclerosis, stroke, fatty liver disease, blindness, and others. Our current research investigates the role of inflammatory cells, adipocytes, Toll-like receptors (TLR), and cytokines in the metabolic syndrome. We have studied two mouse models of impaired hepatic insulin clearance leading to insulin resistance. In the L-SACC mouse model, CEACAM1, a molecule necessary for liver insulin receptor endocytosis, contains a point-mutation which abolishes an essential phosphorylation site (S503A). This dominant-negative CEACAM1 is targeted to the liver only. We have also examined CEACAM1 knock-out (Cc1 KO) mice, wherein CEACAM1 is absent from all tissues. We have induced obesity by feeding L-SACC, Cc1 KO, and wild-type mice either a high-fat or high-fat / high carbohydrate (Western) diet and compared the effects to regular diet-fed mice. We have observed dysregulated TLR, cellular, and cytokine responses by qPCR, serum analysis, and histological methods. We conclude that these mouse models of impaired insulin clearance will be valuable tools for investigating abnormal cellular responses in obesity and type 2 diabetes. This research is supported by the United States Department of Agriculture.
Nuclear factor-kappaB provides mechanistic link between inflammation and prostate cancer (93.8)
Authors: Vykhovanets Eugene , MacLennan Gregory T , Vykhovanets Olena , Gupta Sanjay 
Published: January 2023

Abstract
The mechanistic link between chronic inflammation and prostate cancer (PCa) is missing, despite accumulating evidence suggests a role for inflammation in the etiology of this disease. We hypothesized that nuclear factor-kappaB (NF-κβ) activation might result in the production of signals for mononuclear cell (MNC) accumulation into the prostate. We used transgenic NF-κB-Luc Tag mice that demonstrate the highest levels of NF-κB-dependent luminescence in vivo and ex vivo at 4 h after i/v IL-1β administration. In these mice, IL-1β-induced NF-κB activation correlated with MNC trafficking, expression of chemokines and IL-17 in the prostate. Next we determined the possibility of IL-17 producing cells to be targets of specific pro-inflammatory signals that initiate inflammation at sites of prostate inflammatory atrophy (PIA) in PCa patients. To immunophenotype the inflammatory cell infiltrates in formalin-fixed paraffin-embedded whole mount radical prostatectomy specimens two-color staining was employed. MNC were the predominant IL-17 producing cells in PIA lesions. The number of IL-17 positive cells in the acini of PIA lesions was 5-fold higher than in PCa areas; and 3-fold higher than around PCa or other pathologic patterns studied. Taken together, our findings support the hypothesis that NF-κB provides a mechanistic link between cell-mediated inflammation, PCa development and its subsequent progression.
Nitric Oxide and Carbon Monoxide mediate the anti-inflammatory effects of Remote Ischemic Preconditioning (RIPC) (94.12)
Authors: simao antonio  felipe , Souza-Filho Marcos  V P , Souto Fabricio Oliveira , Cunha Fernando Queiróz , Ribeiro Ronaldo Albuquerque 
Published: January 2023

Abstract
Remote ischemic preconditioning(RIPC) consists of short periods of ischemia followed by reperfusion, which induces protective anti-inflammatory effect on distance organs. Studies have shown that inflammatory parameters such as leukocyte migration are significantly reduced in pre-conditioned animals.However, the mechanism involved in this effect is not yet well established. This study evaluated if NO and CO is mediating the anti-inflammatory activity of RIPC.RIPC model is based on turnstile in right leg of a mouse for 10 minutes followed by 30 of reperfusion. Participation of NO and CO was investigated using inhibitors of iNOS (1400W; 3mg/kg or Aminoguanide; 50mg/kg) and HO-1 (ZnPPIX;10mg/kg) as pre-treatment 30 minutes before RIPC. Afterwards, inflammatory reaction was induced by intraperitoneal administration of Carragenin(500µg/cav).Four hours after the peritoneal cavities were washed with saline and number of migrated leukocytes was evaluated. Chemotaxis ability of neutrophils obtained from mice under RIPC treated or not with the inhibitors was analyzed in Boyden Chamber. Expression of CXCR2 and GRK2 in neutrophils was determined by flow cytometry and immunofluorescence, respectively.CO and NO inhibitors prevented the inhibitory effect of RIPC upon Cg-induced neutrophil migration into peritoneal cavity of animals, as well as, upon KC induced neutrophils chemotaxis in Boyden Chamber. The reduction of neutrophil chemotaxis by RICP correlated with CXCR2 internalization and increase of GRK2 expression in the neutrophils.CO and NO production prevented these effects of RICP.Thus,inhibitors of HO-1/CO and NOSi/NO pathways seem to mediate the anti-inflammatory effect of RICP.
A novel method to evaluate inhibitors of cell adhesion using physiological conditions generated using microfluidic flow cells (94.14)
Authors: Conant Carolyn G , Schwartz Michael , Ionescu-Zanetti Cristian 
Published: January 2023

Abstract
Development and assessment of cell adhesion inhibitors are fundamental to allergy, asthma, clotting, neurodegeneration, and cancer drug research and discovery. Here, we present a cell-based screen of adhesion inhibitors directed against VLA-4, a molecule found on the surface of most leukocytes and key to the inflammatory process. Reduction of VLA-4 mediated adhesion to the ligand, VCAM-1 , was evaluated using a micro-scale flow cell system designed to run automated shear flow protocols for cell biology experiments in high throughput. The screen was performed under physiologically-relevant shear flow conditions of 0.8 dyn/cm2. The flow was also exploited to streamline the experimental workflow and mitigate effects of inconsistent washing as is the case with such experiments performed in microplates using pipet based washing. The time to perform the entire experiment from coating the plate with recombinant VCAM-1 to manual data collection was reduced to under 3 hours compared to 10 hours for a standard microplate adhesion inhibition assay. Cell adhesion images were captured using a standard inverted microscopy setup and data were analyzed using proprietary software. Using three inhibitors against VLA-4, we showed a dose-dependent decrease in cell adhesion in two of the three compounds leading to a complete block of adhesion. IC50 values were determined to be 600pM and 1/32000 respectively. The third compound's IC50 was 10nM although complete inhibition was never observed; as well, homotypic cell aggregation was observed at a concentration of 1nM. Homotypic aggregation would not typically be observed in a microtiter plate adhesion assay screened for bulk fluorescence in a plate reader.