bluuebunny/crossref_metadata_2025_split huggingface dataset:
Biomarkers of food intake and nutrient status are associated with glucose tolerance status and development of Type 2 diabetes
Authors: Savolainen Otto , Lind Mads Vendelbo , Bergström Göran , Fagerberg Björn , Sandberg Ann‐Sofie , Ross Alastair 
Published: November 2023

BACKGROUNDDiet is frequently associated with both the development and prevention of type 2 diabetes (T2D) but there are a lack of objective tools for assessing the causal relationships between diet and T2D. Biomarkers of dietary intake could help strengthen the link between a healthy diet and prevention of diabetes.OBJECTIVEThe objective of this study was to explore how diet is related to glucose tolerance status (GTS) and future development of T2D irrespective of metabolic syndrome (MetS) risk factors, using dietary biomarkers as an objective measure of dietary intake unconfounded by recall and reporting bias.RESEARCH DESIGN AND METHODSDietary biomarkers were measured in plasma from 64‐year old women with different glucose tolerance classifications (normal glucose tolerance; NGT (n=190), impaired glucose tolerance; IGT (n=209), and diabetes (n=230)), randomly selected from the population register in Gothenburg, Sweden. The same subjects were followed up after 5 years to determine changes in glucose tolerance (NGT (n=167), IGT (n=174) and diabetes (n=159)). Analysis of covariance (ANCOVA) adjusted for significant measures of MetS was used to explore baseline data for associations between dietary biomarkers, GTS and new T2D cases at follow up (n=69).RESULTSAfter adjustment for MetS risk factors, alpha‐tocopherol, alkylresorcinols C17 and C19 (markers of whole grain wheat and rye), b‐alanine (meat), eicosapentaenoic acid (fish) and linoleic acid were associated with GTS and 3‐carboxy‐4‐methyl‐5‐propyl‐2‐furanpropanoic acid (CMPF) (fish) and alpha‐tocopherol with future development of T2D.CONCLUSIONSSeveral dietary biomarkers were strongly associated with GTS irrespective of MetS factors, underlining the role of diet in development and prevention of T2D. The use of multiple dietary biomarkers can provide a link with diet that is unencumbered by recall bias normally associated with dietary studies and allows examination of the role of diet even when dietary information is not available.
A Role of IL‐21 in Pulmonary Fibroblast Activation
Authors: Sathiaseelan Roshini , Huang Chaoqun , Senavirathna Lakmini Kumari , Liu Lin 
Published: November 2023

Idiopathic pulmonary fibrosis (IPF) is a progressive, fatal interstitial fibrotic lung disease with a poor prognosis. Activated fibroblasts produce excess extracellular matrix leading to fibrosis. Transforming growth factor β (TGFβ) is increased in IPF and is one of the most important factors for the activation of fibroblasts. T lymphocytes‐derived cytokines such as IL‐13 and IL‐4 is known to stimulate macrophages to secrete TGFβ. However, the direct effects of these cytokines on the activation of fibroblasts are poorly understood. In this study, we investigated the role of IL‐21 in pulmonary fibroblast activation. RNA sequencing analysis revealed that TGFβ1 increased the expression of IL‐21 receptor (IL‐21R) in primary human pulmonary fibroblasts (HPF). The up‐regulation of IL‐21R was confirmed by real‐time PCR in normal pulmonary fibroblasts (HPF, CCD13, CCD8) and IPF fibroblasts (LL97A and LL29). To determine the effects of IL‐21 on the fibroblast functions, pulmonary fibroblasts were serum‐starved for 16 hours and then treated with TGFβ1 and/or recombinant human IL‐21 for 2 days. The mRNA expression of myofibroblast marker, α‐smooth muscle actin was markedly increased by TGFβ and IL‐21 in all of the fibroblast cell lines. We conclude that IL‐21 promotes the differentiation of fibroblasts to myofibroblasts via TGFβ‐induced IL‐21R expression.Support or Funding InformationR01HL116876 and P20GM103648
Capsaicin Synergizes with Camptothecin to show Enhanced Apoptotic Activity in Small Cell Lung Cancer
Authors: Friedman Jamie Rae , Perry Haley E , Brown Kathleen C , Akers Austin T , Nolan Nicholas A , Rollyson William D , Chen Yi Charlie , Denning Krista L , Brown Linda G , Dasgupta Piyali 
Published: November 2023

Platinum‐based chemotherapy is the cornerstone of small cell lung cancer (SCLC) treatment. Chemotherapy initially shows excellent outcomes with 60–80% patients achieving remission. However, patients relapse within a year and after that the tumor does not respond to chemotherapy or radiation. There is only one FDA‐approved drug for relapsed SCLC, which is camptothecin (topotecan). It has been shown to display an objective response rate of about 3%. Therefore agents which can increase the response rate of camptothecin should be of considerable benefit to SCLC patients. We tested the combinatorial effects of the nutritional compound capsaicin with camptothecin in human SCLC. Caspase‐3 activity assays reveal that the combination of camptothecin and capsaicin displayed greater apoptotic activity than any of the compounds used alone. Statistical analysis using the Chou‐Talalay isobologram showed that capsaicin displayed synergistic apoptotic activity with camptothecin. Chicken chorioallantoic membrane assays confirmed the combinatorial effects of capsaicin and camptothecin. The synergistic effects of capsaicin and camptothecin are mediated by the calpain pathway and involve the proteolytic cleavage of the pro‐apoptotic protein Bax to generate a more potent pro‐apoptotic Bax fragment protein. The results of our studies will foster the hope of novel second‐line treatment regimens for human SCLC.Support or Funding InformationFunding for our study was supported by an NIH R15‐AREA Grant (2R15CA161491‐02) and a WVU‐MU Health Partnership Grant.
Loss of function mutation in TIMP2 gene accelerates tumorigenesis and mortality in murine model of lung cancer through EGFR signaling
Authors: KUMAR SARVESH , Jensen Sandra , Peeney David , Chowdhury Ananda , Wei Beiyang , Stetler‐Stevenson William G. 
Published: November 2023

The aim of this study is to determine the effect of the tissue inhibitor of metalloproteinase‐2 (TIMP‐2) on lung tumorigenesis and investigate the underlying molecular mechanisms using an orthotopic mouse model with a loss of function mutation in the Timp2 gene (T2M). T2M and wildtype (WT) control mice were given 1×106 Lewis lung carcinoma cells transfected with luciferase (LL/2‐Luc‐M38, Caliper) in 50μL PBS via intratracheal installation. IVIS imaging revealed a higher tumor burden in T2M mice compared to WT littermates, suggesting that loss of function of the Timp2 gene enhances tumor growth (p<0.05). We also conducted a Kaplan‐Meier analysis to determine the effect of this mutation on mortality following cancer development. We found that LL/2 tumor‐induced mortality was significantly higher in T2M mice compared to WT controls (p = 0.01). Histologic analysis and H&E staining of lung tissue sections revealed a significant increase in the number of tumor nodules of T2M mice compared to WT controls (p <0.01). CD31 staining showed a significant increase in microvessel density (MVD) in T2M mice compared to WT controls (p<0.01). Given that VEGF is a primary driver of tumor neovascularization, we determined mRNA levels of VEGF expression in healthy and tumor bearing mice by qRTPCR analysis. Interestingly, basal VEGF expression levels were increased in lungs of both non‐tumor bearing and tumor bearing T2M mice compared to WT controls (p<0.01). VEGF is the direct downstream target of HIF‐2a, a transcription factor implicated in tumor hypoxia and expression correlates with decreased overall survival in non– small cell lung cancer (NSCLC) patients. Similarly, an examination of TIMP‐2 and HIF‐2a mRNA expression in a small cohort of patient samples revealed decreased TIMP‐2 and increased HIF2a in NSCLC tumors compared to surrounding non‐tumor lung tissue. Hypoxia induces EGFR signaling through HIF‐2a. Thus, we determined the effect of mutated T2M on the EGFR signaling pathway. We found increased levels of EGFR phosphorylation as well as downstream ERK and Akt activation in tumor bearing and non‐tumor bearing T2M lungs compared to WT controls. In conclusion, these findings offer new avenues for TIMP‐2 research in regulating hypoxic mediators within the tumor microenvironment through EGFR signaling, suggesting TIMP‐2 as a novel bio‐therapeutic for lung cancer therapy.Support or Funding InformationThis work is supported by center for cancer research, intramural research program, NCI/NIH research grants ZIA BC011204 & ZIA SC 009179 to WGSS
Whole Slide Quantitative Image Analysis Can Demonstrate Bleomycin Induced Dose‐Dependent Increases in Fibrotic Areas Within Histologically Stained Rat Lung Tissue
Authors: Young Alan , McElroy Mary , Naylor Stuart , Anderson Mark , Bigley Alison , McGregor Kieran , Sherry Lorcan 
Published: November 2023

RationaleThe accurate classification and quantification of specific morphological and cellular components within the bleomycin (Bleo)‐induced rodent lung tissue model is challenging. This study aimed to employ histology in combination with whole slide digital image analysis to quantify the extent of lesion formation and content within lung tissue from Bleo treated rats.MethodsMale SD rats, n=5 per group (Gr), were treated with either saline (Gr1) each day for 7 days or 1.5mg/Kg Bleo (Gr2) and 3.0mg/Kg Bleo (Gr3) on one dose occasion or 0.5mg/Kg Bleo (Gr4) and 1.0mg/Kg Bleo (Gr5) each day for 7 days. On day 22, lungs were fixed, paraffin embedded, sectioned and histologically stained using Picrosirius Red. Whole slide images of lung tissue were captured and analysis algorithms developed by OracleBio (using Definiens Tissue Studio® software) to define specific regions of interest (ROI) per section including Parenchyma and Lesion, excluding basal collagen. Collagen content and cell density can then be quantified within the Lesion and Parenchyma ROI, but have not been presented in this abstract. The % Fibrotic Lesion area was calculated relative to the combined Lesion and Parenchyma areas only.ResultsThe mean % Lesion area in Gr1 was 1.1% (95% CI: 0.5 to 2.7). This value was attributed as artefact which was classified during the objective application of the analysis algorithm and was excluded from further analysis. There was a clear trend showing an increase in % Lesion area as the Bleo dose increased from 0.5 to 3.0mg/Kg. The mean % Lesion area in Gr2 was 7.0% (95% CI: 4.5 to 9.6), Gr3 was 9.1% (95% CI: 1.7 to 16.5), Gr4 was 10.1% (95% CI: 6.5 to 13.7) and Gr5 was 16.2% (95% CI: 9.4 to 23.1). A significant difference was observed between Gr2 and Gr5 (p = 0.0124).ConclusionQuantitative whole slide image analysis can be utilised to accurately classify therapeutically relevant regions of interest. This may provide a new tool to assess and better interpret changes in morphological and cellular tissue parameters within histologically stained lung tissue sections from pre‐clinical bleomycin fibrosis models.Support or Funding InformationCharles River Laboratories and OracleBio
Inflammasome Inhibition in Influenza A Virus Infected Juvenile Mice Leads to Improved Survival and Outcomes
Authors: Ravindran Nandini , Koch Clarissa M , Ridge Karen M , Coates Bria M 
Published: November 2023

RationaleInfluenza A virus (IAV) is a highly contagious respiratory virus that infects up to 40% of children each year. Young children who progress to lower respiratory tract IAV infection are at risk of severe, life‐threatening disease. We previously demonstrated that viral clearance does not predict severity of illness in juvenile mice infected with IAV. Instead, excess activation of the NLRP3 (NLR family pyrin domain containing 3) inflammasome by IAV and the resultant cytokine storm are associated with increased lung injury in juvenile mice. Therefore, we hypothesized that inhibition of the NLRP3 inflammasome would protect juvenile mice from lethal IAV infection.MethodsJuvenile mice (4 week‐old) were intratracheally infected with IAV (WSN 12.5 PFU). Two, three, and four days following infection, treatment with either an NLRP3 inflammasome inhibitor (MCC950) or saline control was initiated and continued until resolution of infection or death. Mice were evaluated for viral clearance and the development of acute lung injury. In addition, production of pro‐inflammatory cytokines, antiviral interferons and activation of the NLRP3 inflammasome were measured in bronchoalveolar lavage fluid (BALF). The immune cell profile was also evaluated in response to MCC950 treatment.ResultsInhibition of the NLRP3 inflammasome with MCC950 3 days following infection protected juvenile mice from IAV‐induced lung injury and significantly increased survival. Earlier or later inhibition was not protective. Compared with vehicle‐treated controls, MCC950‐treated mice achieved an equal rate of viral clearance, but secreted decreased amounts of pro‐inflammatory cytokines. Further studies will be conducted to assess the effects on the immune cell population.ConclusionModulation of the juvenile inflammatory response to IAV does not impair viral clearance. Instead, inhibition of excessive inflammation in juvenile IAV infection with a specific NLRP3 inflammasome inhibitor attenuates lung injury and improves survival in IAV‐infected juvenile mice. Our data suggest that therapies directed at the inflammatory response to IAV, and not viral clearance, warrant further investigation for treatment of severe IAV infection in children.Support or Funding InformationHL128194 and HL071643
The Effect of Plumbagin on Proinflammatory Cytokines Expression in LPS‐Activated BV‐2 Microglia Cells
Authors: Messeha Samia S , Zarmouh Najla O , Kolta Malak G , Soliman Karam F. 
Published: November 2023

In the central nervous system, activated microglial cells produce proinflammatory mediators such as inducible nitric oxide (NO2) and cytokines. Uncontrolled release of these mediators can cause neuronal damage in the brain leading to neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Therefore, inhibition of these proinflammatory mediators may provide a neuroprotective strategy in preventing neurodegenerative diseases. the other hand, plumbagin is a biologically active compound in the roots of the Indian medicinal plant Plumbago zeylanica L. which has been used in treating rheumatoid arthritis. The present study was conducted to examine the antiinflammatory effect of the plumbagin in LPS‐activated microglial BV‐2 cell model. The obtained data indicated that plumbagin (IC50 = 0.386 uM) was about 20‐fold more potent than l‐N6‐(1‐iminoethyl) lysine (LNL), the selective inhibitor of inducible nitric oxide synthase (iNOS) (IC50 =7.4 uM). Moreover, our immunofluorescence study and ELISA confirmed the positive relationship between NO2 decrease and iNOS expression inhibition. RayBio AAM‐CYT‐3 and 4 cytokine antibody protein arrays were selected for detecting extracellular proinflammatory cytokines expression changes in LPS–activated BV‐2 cells in the presence of plumbagin. In the activated BV‐2 cells, 2 ums of plumbagin significantly inhibited the release of IL‐6, MCP 1, MCP5, MIP‐1 alpha and gamma, IL‐1 alpha, G‐CSF, IL‐12 p40/70 and MDC. The obtained results of ELISA analysis validated the cytokines array data. Taken all together, we concluded that plumbagin is a potent antiinflammatory agent reduces multiple proinflammatory cytokines and iNOS.Support or Funding InformationSupported by NIH Grants G12 MD007582 and P20 MD 006738
West‐Nile virus replicon particles infect 293T cells expressing Dendritic Cell‐Specific Intercellular Adhesion Molecule‐3‐Grabbing Non‐Integrin Related.
Authors: Leonard Nathan , Crowson Nicole , Madigan Amanda , Schmitz Madilyn , Alcala Elana , Martinez Osvaldo 
Published: November 2023

West‐Nile virus (WNV) is an arthropod‐borne virus that is frequently transmitted to humans through the bite of an infected mosquito. Less than one percent of cases result in a serious neurological illness such as meningitis or encephalitis. To investigate WNV tropism we have established several 293T stable cell lines expressing dendritic cell‐specific intercellular adhesion molecule‐3‐grabbing non‐integrin related (DC‐SIGNR) or DC‐SIGN. Previous studies have suggested that WNV infection efficiency is enhanced by expression of DC‐SIGNR, but not DC‐SIGN on target cells. We demonstrate high levels of DC‐SIGNR expression in selected 293T cells. Replication incompetent virus replicon particles (VRPs) are used to safely and conveniently study viral entry. Successful VRP entry into target cells transfers a mutated incomplete viral genome containing the GFP gene which is expressed from the infected cells. Infections assays using WNV (NY99) VRPs as well as a variant of WNV (NY99) which contains the beta‐lactamase enzyme, show significant infection in DC‐SIGNR expressing, but not control cells that do not express DC‐SIGNR. The establishment of this entry assay system will allow us to safely study virus tropism.Support or Funding InformationStudent Research Grants, Winona State University Winona, MN
RGDSK Peptide Functionalized Helical Rosette Nanotubes (RGDSK‐HRNs) Block Integrin αvβ3 and Inhibit E. coli Adherence to Intestinal Porcine Epithelial 1 Cell Line (IPEC1) in vitro
Authors: Le Nguyen Phuong Khanh , Quach Chi Cuong , Aulakh Gurpreet , Gerdts Volker , Fenniri Hicham , Singh Baljit 
Published: November 2023

E. coli infection is a leading cause of intestinal disease and death, and inflicts billions of dollars in economic losses. There is ongoing effort to find ways to reduce intestinal colonization by E. coli. Integrin αvβ3, recognizing arginine‐glycine‐aspartic acid (RGD) sequences, has important functions in cell adhesion, signaling and survival. However, the expression and role of integrin αvβ3 in the adhesion of E. coli on intestinal epithelium remain unknown.We used porcine jejunum, IPEC1 cells and E. coli K88 to firstly explored the hitherto unknown expression of integrin αvβ3 with light, fluorescent and electron immunocytochemistry, immunoprecipitation and Western blots. We then used RGDSK peptide and RGDSK‐HRNs to test the role of integrin αvβ3 in adhesion of E. coli K88 to IPEC1 cells.Normal porcine jejunum strongly expresses integrin αvβ3 on the nucleus and apical surface of epithelium and gland cells. The expression of integrin αvβ3 decreased in the epithelium but increased on vascular endothelium of the jejunum infected with E. coli or E. coli associated with Salmonella. Integrin αvβ3 was found on plasma membrane, cytoplasm, and nucleus of IPEC1 cells. The expression of integrin αvβ3 on IPEC1 decreased at 15 minutes but returned to normal after 90 minutes of infection with E. coli K88 (P<0.05). Immuno‐gold quantification showed changes in sub‐cellular expression of the integrin αvβ3 in IPEC1. The light, fluorescent and electron microscopy, and western blots showed novel expression of integrin αvβ3 on E. coli K88. Finally, we found that RGDSK peptides and RGDSK‐HRNs bind integrin αvβ3 on both IPEC1 and E. coli K88. RGDSK peptides block the attachment of E. coli to IPEC1 (5 minutes IPEC1 exposed to 50μM RGDSK, P<0.05), and reduce survival of E. coli (10 minutes IPEC1 exposed to 50μM RGDSK, P<0.05). Dose dependent RGDSK‐HRNs mediates the attachment of E. coli to IPEC1 more efficiently and regularly (10 minutes IPEC1 exposed to HRNs containing equivalent 5μM RGDSK, P<0.001). These are the first data to show a role for the integrin αvβ3 in the attachment of E. coli to the intestinal epithelium, and that novel RGDSK helical rosette nanotubes can inhibit the attachment of E. coli to the epithelium.Support or Funding InformationSaskatchewan Agriculture Development Fund and NSERC
Development of an in vitro Bladder Epithelial Cell Culture System
Authors: Kim Seongmi 
Published: November 2023

Urinary tract infections (UTIs) are very common and highly recurrent. Our laboratory has developed a murine model of the recurrent bladder infection: C3H/HeN mice with a history of UTI that was cleared with antibiotic therapy become “sensitized” to severe recurrent UTI upon subsequent UPEC challenge, while “resolved” mice are resistant to challenge infection relative to age‐matched “naïve” mice. Quantitative proteomics has found evidence of bladder epithelial (urothelial) cell remodeling in sensitized mice. To study these remodeling changes ex vivo, we have adopted a primary epithelial cell culture method for use with urothelial cells. Urothelial cells isolated from naive, resolved and sensitized mice were passaged in matrigel‐suspended “spheroid” culture with conditioned media from L‐cells expressing Wnt, R‐spondin and noggin to promote proliferation, then cultured on transwells, allowing them to differentiate into polarized urothelial layers with terminal differentiation markers. We observed sensitized urothelium had characteristic differences in its morphology and cytokine response to UPEC infection that were similar to differences observed in vivo. Serum biomarkers that predict chronic cystitis and predisposition to rUTI in human and mice, such as IL‐6, G‐CSF, and CXCL1 (KC), showed increased expression patterns in sensitized urothelia. Interestingly, baseline chemokine RANTES expression is suppressed in sensitized urothelia. These results indicate that mucosal remodeling is happening at the urothelial cell level, and thus there is potential to study these changes in great detail in vitro. This primary culturing system represents a significant advance in the study of bladder mucosal responses and has great potential for translation to human clinical studies.Support or Funding InformationMucosal Immunology Studies Team consortium U01 AI095542