bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of cellular stress on retinoid homeostasis in the small intestine
Authors: Zalesak‐Kravec Stephanie , Yu Jianshi , Pilli Nageswara , Kane Maureen A. 
Published: May 2022

Vitamin A is an essential diet‐derived nutrient. Retinoids are compounds with Vitamin A activity and are essential for many biological processes, including proper cell function and tissue organization. The main active metabolite of Vitamin A is all‐trans retinoic acid (RA), which is critical for nuclear receptor mediated signaling for cell proliferation, differentiation, and apoptosis. Retinoid signaling and RA are essential for intestinal immune homeostasis. Intestinal epithelial cells (IEC) produce high levels of RA, which is crucial for RA‐dependent signaling, lineage commitment, and barrier function. Vitamin A deficiency can alter intestinal immune and IEC homeostasis.There are many mechanisms in place to maintain retinoic acid homeostasis, including the expression of the main intracellular chaperone protein cellular retinol binding protein, type 1 (CRBP1). CRBP1 binds to retinol and retinal, protects them from non‐specific oxidation, and facilitates their delivery to the appropriate enzymes for RA biosynthesis. CRBP1 has been shown to be decreased in diseases that display dysfunctional proliferation and differentiation, including cancer, inflammatory disorders, fibrosis, and infection. Reduction of CRBP1 directly correlates with reduction in the active metabolite, RA, and restoration of CRBP1 expression has been shown to increase RA levels and positively impact RA‐dependent outcomes. Therefore, monitoring CRBP1 and retinoid metabolite levels in response to cellular stress will provide useful information about the effect of CRBP1 loss on retinoid signaling.While previous studies have explored the effect of cellular stress on RA levels, the effect of stress on the entirety of the retinoid signaling pathway is not well understood. The objective of these studies is to systematically determine the effect of cellular stress on retinoid homeostasis.We have established three models of cellular stress, including hypoxia, oxidative stress, and inflammation, in two human intestinal epithelial cells lines, HIEC‐6 and Caco‐2. To mimic hypoxia, cells are treated with 0‐100 μM CoCl2 for 24 hours. To mimic oxidative stress, cells are treated with 0‐100 μM H2O2 for 24 hours. To mimic inflammation, cells are treated with 0‐1 μg/mL lipopolysaccharide for 48 hours. These conditions were chosen because there was no effect on cell viability. CRBP1 and RA levels will be measured through targeted mass spectrometry methods developed by our lab. Additionally, expression levels of genes associated with retinoid metabolism will be measured. Our current data shows that RA levels decreased in response to hypoxia, oxidative stress, and inflammation in both HIEC‐6 and Caco‐2 cells.
Nigella Sativa Can Reverse Statin‐Induced Renal Injury in Rats: An Ultrastructural Study
Authors: Oules Jon , Thomann Charity , Youakim Magdy , Amin Shaimaa , Hassan Sherif 
Published: May 2022

Introduction and HypothesisAtorvastatin (ATO) and its siblings in the statin drug class are widely prescribed as a lipid lowering agent. However, ATO in high doses can induce renal toxicity via oxidative stress. Thymoquinone (TQ), an extractive of Nigella sativa,is a natural antioxidant that has been shown to protect the kidney via its anti‐inflammatory effects. The aim was to study the anti‐inflammatory effect of posttreatment TQ and whether it could reverse the ATO‐induced oxidative injury to the rat kidney.MethodsWe used 4 groups of adult male Sprague Dawley rats (control; TQ‐treated; ATO‐post treated; ATO‐ post treated plus TQ‐treated). Each group was formed of 10 rats. Ultrastructural, and morphometric analyses of the rat kidneys were performed. Statistical analysis was done using SPSS. One‐way analysis of variance (ANOVA) and post hoc tests (Bonferroni test) were used to compare the means of different groups, expressing data as mean ± standard deviation (SD) and P values of less than 0.01 were expected to be significant.ResultsMorphometric analyses of the ultrastructural examination revealed that the histopathological changes induced by post treatment ATO has improved in the ATO/TQ‐treated group compared to the ATO‐ post treated group.ConclusionPosttreatment TQ appears to reduce and reverse renal injury due to ATO‐induced oxidative stress. TQ can potentially play a role in alleviating side effects in patients with kidney disease who are being treated with high dose of atorvastatin.
HH pathway orchestrates the epithelial‐mesenchymal crosstalk during human lung branching morphogenesis
Authors: Belgacemi Randa , Danopoulos Soula , Deutsch Gail , Dormoy Valerian , Al Alam Denise 
Published: May 2022

RationaleHedgehog (HH) pathway plays an essential role in lung branching morphogenesis in rodents. However, little is known about the role of this pathway in human lung development. We hypothesize that HH pathway is necessary for branching during human lung development and hence in pulmonary hypoplasia.MethodsExpression of HH pathway components was analyzed in single cell and bulk RNA‐sequencing data from publicly available lung datasets. Fresh human fetal lung tissues between gestational ages 10‐18 weeks were collected. RT‐qPCR, fluorescent in situ hybridization (FISH) and immunofluorescence staining (IF) were performed to determine spatiotemporal localization of HH pathway actors. In addition, distal lung segments were cultured in air‐liquid interface for up to 48 hours in the presence or absence of HH inhibitor (5E1). RT‐qPCR, western blot and IF were performed to investigate the effect of HH pathway inhibition on epithelial‐mesenchymal markers and associated signaling pathways.ResultsBulk and scRNA‐seq analysis showed an early expression of HH pathway components during human lung development. We noted an inverse expression between activators and inhibitors receptors (PTCH1/HHIP) over time. PTCH1 had a higher expression at 20 weeks compared to 14 weeks while the opposite was observed for HHIP. HHIP FISH staining was restricted to ACTA2+ airway smooth muscle cells while PTCH1 was primarily localized in mesenchymal populations adjacent to the tip of airway epithelial buds where SHH ligand is produced. Inhibiting HH signaling ex vivo reduced branching during development (2.5‐fold vs control, p<0.01, n=8). Consequently, SOX9expression, an epithelial progenitor marker, was increased whereas ACTA2 was decreased (respectively 1.5‐ and 2‐fold vs control, p<0.05, n=8). These results were confirmed by western blot and IF staining. No change was observed for SOX2 gene expression while a significant increase in protein expression was observed. FGF10 increased following HH inhibition (1.5‐fold vs control, p<0.01, n=8), but the expression of ETV4and 5 decreased (1.5‐fold vs control, p<0.05, n=8). Finally, we demonstrated that HH signaling elements (including SHH, PTCH1, GLI2) were downregulated in lung tissues from congenital diaphragmatic hernia patients.ConclusionWe have determined for the first‐time gene expression and localization of HH pathway during human lung morphogenesis. Moreover, we showed that HH signaling inhibition alters important genes and proteins expression, such as epithelial‐mesenchymal progenitors and associated pathways, required for proper branching processes. Understanding the role of HH pathway on human lung development could lead to the identification of novel therapeutic targets for childhood pulmonary diseases.
Aerobic Fitness as a Predictor of Conduit, Resistance and, Microvascular Arterial Function
Authors: Hibner Brooks A. , Lefferts Elizabeth C. , Lima Natalia S. , Sherman Sara R. , DeJonge Sydney R. , Baynard Tracy , Fernhall Bo 
Published: May 2022

IntroductionVascular dysfunction increases cardiovascular disease (CVD) risk, whereas aerobic fitness (VO2peak) improves arterial function and decreases CVD risk. However, it is unclear if VO2peak has the same degree of importance across each segment of the arterial tree (e.g., conduit, resistance, and microvasculature). This is relevant given that research has not assessed these three segments in the same cohort.PurposeTo examine the relationship of VO2peak and conduit, resistance, and microvasculature arterial function.MethodsGraded exercise testing assessed VO2peak in 33 males (41±19 years, 25.6±2.6 kg/m‐2, 41.1±13.6 ml/kg‐1/min‐1) and 28 females (30±12 years, 23.3±4.3 kg/m‐2, 33.6±8.5 ml/kg‐1/min‐1). Concurrent brachial artery flow‐mediated dilation (FMD) and forearm near‐infrared spectroscopy (NIRS) examined conduit, resistance, and microvascular arterial function. FMD measured with ultrasonography to continuously record arterial diameters and Doppler velocities for 1 min baseline, 5 min forearm occlusion (250 mmHg), and 3 min recovery. The maximum percent change in diameter compared with baseline defined FMD (%) and was used to evaluate conduit arterial function. Reactive hyperemia (RH) of forearm resistance arteries was calculated as peak brachial blood flow following cuff release. Forearm tissue saturation index using NIRS during FMD was used to evaluate microvasculature function (reperfusion slope and reperfusion magnitude). Multivariate regression analysis was performed using VO2peak, age, sex, and BMI as independent variables.ResultsAge was the only predictor of FMD (β ‐0.48 [‐0.02, ‐0.01], p=0.001), sex was the only predictor of RH, females exhibited lower RH, (β ‐0.66 [‐0.30, ‐0.15], p<0.001), and VO2peak only predicted reperfusion slope (β 0.56 [1.46, 4.52], p< 0.001) and reperfusion magnitude (β 0.36 [3.15, 41.66], p=0.02).ConclusionIn our cohort, aerobic fitness was associated with microvascular function when age, BMI, and sex were included in the model. Only age was associated with conduit artery function, and only sex was associated with resistance arterial function. Each segment of the arterial tree was affected differently by VO2peak, age, and sex. These data show the importance of including several potentially important variables when investigating the association between VO2peak and arterial function.
Spatial Whole Transcriptome Profiling of the Tumor Microenvironment in Prostate Carcinomas
Authors: Walker Dan , Giangarra Valeria , Anaparthy Naishitha , Williams Stephen , Sheldon Jyoti , Turkekul Mesruh , Mielinis Paulius , Gallant Caroline , Chell James , Taylor Sarah 
Published: May 2022

BackgroundDespite years of studies and effort, the best strategies for treating prostate cancer and minimizing the complications of treatment remain unanswered questions. This gap in knowledge is partially due to the inability to dissect the complex heterogeneous tumor microenvironment (TME) and immune compartment. Spatially resolved molecular profiling of tumor sections will enhance our understanding of these complexities; However, it has been particularly challenging to do spatial molecular profiling in formalin‐fixed paraffin‐embedded (FFPE) tissues due to RNA degradation associated with this tissue‐embedding method, which is routinely used in oncology workflows.The 10x Genomics Visium Spatial Gene Expression Solution for FFPE tissue overcomes these limitations, enabling spatial gene expression analysis of FFPE tissues combined with classical histology staining techniques such as Hematoxylin & Eosin (H&E) staining and immunofluorescence.MethodsWe used the 10x Genomics Visium Spatial Gene Expression Solution for FFPE tissue to analyze and resolve tumorigenic profiles in sections of normal and adenocarcinoma prostate samples. This assay incorporates ~5,000 molecularly barcoded, spatially encoded capture probes in spots over which the tissue is placed, imaged, and permeabilized. Imaging and sequencing data are processed together, resulting in a spatially resolved transcriptional readout.ResultsWe profiled the whole transcriptome in normal, invasive adenocarcinoma, and acinar cell carcinoma FFPE human prostate tissues.Unsupervised clustering of the whole transcriptome data from normal, invasive adenocarcinoma, and acinar cell prostate carcinoma FFPE sections enabled the identification of 2 different regions, which had a well defined spatial distribution within the tissues. Well known prostate gland and prostate‐cancer markers were over‐expressed in the corresponding healthy and cancerous portions of the tissue, validating the performance of this method. We found that, while in healthy tissues basal cells and luminal cells are spatially organized, this pattern is lost in tumor samples, where luminal cells are greatly expanded in the invasive carcinoma region and do not colocalize with basal cells. Moreover, T lymphocytes are dispersed throughout the whole tissue section in the adenocarcinoma, while plasma B cells are located in the peritumoral region which could impact prognosis.ConclusionsSpatial whole transcriptome analysis opens new opportunities for better understanding the TME which can not only help discover novel predictive tumor biomarkers, but also enable identifying cell type and tumor region specific drug targets.
Assessing the Antinociceptive Effects and Tolerance Development of Decursinol in Nociceptive, Inflammatory, and Neuropathic Pain
Authors: Crawford LaTaijah , Henderson‐Redmond Angela , Kim Sangyub , Karelia Deepkamal , Sepulveda Diana , Lu Junxuan , Morgan Daniel 
Published: May 2022

Chemotherapy‐induced neuropathic pain is a prevalent side effect of treatment with anti‐neoplastic agents. This debilitating neuropathy is characterized by excessive pain resulting in mechanical and sensory hypersensitivity. There is a critical clinical need to develop non‐opioid alternatives to manage chronic pain. A potential alternative for the management of pain is decursinol, a pyranocoumarin compound isolated from the root of the Angelica gigas Nakai (AGN) plant. Pharmacokinetic studies find that clinically, AGN is first metabolized into decursin (D) and decursinol angelate (DA) which are subsequently metabolized into decursinol (DOH). D and DA possess anti‐inflammatory, neuroprotective, antinociceptive, and antioxidant modulatory properties. Studies have reported acute analgesic properties of DOH, but tolerance development along with its impact on chronic pain remains unknown. Therefore, the goal of this study was to assess the antinociceptive effects and potential tolerance to once‐daily injections of DOH (50 mg/kg) in models of thermal (hot plate and tail‐flick), inflammatory (formalin test), and chronic (cisplatin‐induced neuropathic; CINP) pain. We hypothesized that DOH has prolonged antinociceptive and anti‐inflammatory effects. Antinociception and tolerance were assessed in male mice treated once‐daily with either vehicle, DOH (50 mg/kg), or morphine (10 mg/kg; formalin only) intraperitoneally (IP) 30 minutes prior to testing for 14 consecutive days. In the formalin assay, following pretreatment, mice received an intraplanar injection of 2.5% formalin into the right hind paw and nocifensive behaviors were recorded and scored. CINP was induced following four weeks of once‐weekly IP injections of cisplatin (5 mg/kg). Following pretreatment, mice with CINP were assessed for mechanical allodynia via an electro‐von‐Frey anesthesiometer. Daily treatment with 50 mg/kg DOH induced prolonged antinociceptive and anti‐inflammatory responses in all nociceptive tests.Evidence of tolerance to DOH in acute thermal nociceptive tests occurred around day 10 for both hot plate and tail‐flick. Acute administration of DOH eliminated inflammatory pain to an undetectable level in both the acute and inflammatory phases of the formalin test. Further, there is evidence of tolerance by day 14 in the acute (but not the inflammatory) phase of the formalin test. Likewise, tolerance was slower to develop to the antinociceptive effects of DOH compared to morphine in the formalin assay. In mice with CINP, DOH (50 mg/kg) caused a full reversal of mechanical allodynia with evidence of partial tolerance by day 3 with complete tolerance not occurring until day 14. These data indicate that DOH may serve as a potential non‐opioid alternative for the management of pain. Future directions for this study include elucidating the mechanism of action for the antinociceptive effects of DOH.
The Role of PER‐1, ET‐1 Axis in Regulating Endothelial Cell Function
Authors: Jakkidi Nishka R. , Gomes Joshua , Sharma Ravindra K. , Li Shiyu , Gumz Michelle L. , Mohandas Rajesh 
Published: May 2022

ObjectiveResults from major epidemiological studies give evidence to the essential role that dietary salt plays in the development of hypertension. Contrastingly, the mechanisms that mediate individual susceptibility to salt‐sensitive hypertension are unknown. Blood pressure (BP) and sodium reabsorption by the kidneys are regulated by the circadian clock. Consistent with these observations, when mice with global knockout (KO) of the circadian protein period 1 (PER1) are exposed to a high salt diet, they demonstrate increased endothelin‐1 (ET‐1) levels, impaired natriuresis, and hypertension. However, the cumulative sodium balance of PER1 KO mice is not different from control mice. Thus, we hypothesize that PER1 might mediate endothelial cell dysfunction and susceptibility to salt sensitive hypertension.Materials and MethodsHuman Umbilical Vein Endothelial Cells (HUVEC) were transfected with siRNA targeting PER1 or non‐targeting siRNA controls. Endothelial cell function was assessed by measurement of nitric oxide bioavailability, proliferation, and migration. Nitric Oxide (NO) was quantified by DAF fluorescence, proliferation by EZQuant™, and migration by measuring velocity and distance travelled by single cells using time lapse imaging.ResultsNitric oxide bioavailability was increased with PER1 knockdown (Mean fluorescence 14433 ± 1344 vs. 7423 ± 472, p<0.005). Migration velocity and distance were decreased with PER1 siRNA compared to non‐target controls, though not statistically significant (Mean distance 118.9 uM ± 11.39 vs. 138 uM ± 10.88, p=0.2) and (Mean velocity 42.3 uM/sec ± 3.98 vs. 46 uM/sec ± 3.6, p=0.14).There were no differences in proliferation at 24, 48, or 72 hours (Fold change in cell numbers of PER1 siRNA treated cells compared to non‐target controls were 0.87 ± .05 at 24 hours, 0.87 ± .09 at 48 hours, and 0.90 ± .06 at 72 hours, p=NS).ConclusionsOur results suggest that knock down of PER1 increases NO bioavailability and decreases migration of endothelial cells. ET‐1 can increase NO bioavailability via ETB receptors. Experiments are ongoing to delineate the role of ET‐1/ETA and ET‐B in mediating the effect of PER1 on endothelial cell function.
Proteome profiling of a S‐Nitrosoglutathione reductase (GSNOR) null mutant reveals that aldo‐keto reductases form a new class of enzymes involved in nitric oxide homeostasis
Authors: Treffon Patrick , Rossi Jacopo , Gabellini Giuseppe , Trost Paolo , Zaffagnini Mirko , Vierling Elizabeth 
Published: May 2022

Nitric oxide (NO) is a short‐lived gas that acts as a signaling molecule in all higher organisms, including plants. Despite the involvement of NO in multiple plant processes, including germination, root growth and fertility, a basic understanding of the mechanisms by which NO exerts its effects is lacking. NO and its derivatives impact these physiological processes through reversible S‐nitrosation of critical protein cysteines. In cells, regulation of NO‐levels is predominantly achieved by reaction of reactive nitrogen species (RNS) with glutathione (GSH), thereby forming S‐nitrosoglutathione (GSNO), a principal NO reservoir. Mutation of Arabidopsis thaliana S‐nitrosoglutathione reductase (GSNOR) leads to higher intracellular concentrations of S‐nitrosothiols, confirming that the GSNOR reduction of GSNO is a major route of GSNO catabolism in plants and other eukaryotes1. We demonstrate in Arabidopsisthat absence of GSNOR results in differential regulation of proteins involved in chlorophyll metabolism, the general stress response and photosynthesis2. In addition, our proteomic analysis identified a significant increase in proteins that belong to the aldo‐keto reductase (AKR) protein superfamily, AKR4C8 and 9. Since specific AKRs have been linked to NO metabolism in mammals, we expressed and purified ArabidopsisAKR4C8 and 9 and close homologues AKR4C10 and 11 and determined that they have NADPH‐dependent activity in GSNO and S‐nitroso‐coenzyme A (SNO‐CoA) reduction. Plants lacking GSNOR also show increased activity of NADPH‐dependent GSNO reduction, consistent with increased AKR activity2. To address the in vivo role of these AKRs, CRISPR/Cas9‐mediated mutant lines will be generated, and their capacity to metabolize RNS assessed. In addition, by subjecting AKR mutant plants to different abiotic and nitrosative stresses, results will uncover the importance of AKR proteins in NO homeostasis during both optimal growth and stress conditions. Taken together, these data define a new, NADPH‐dependent component of NO metabolism that may be integrated with NADH‐dependent GSNOR activity to control NO homeostasis in plants and other organisms.1. Lee, U., Wie, C., Fernandez, B. O., Feelisch, M. & Vierling, E. Modulation of nitrosative stress by S‐nitrosoglutathione reductase is critical for thermotolerance and plant growth in Arabidopsis. Plant Cell 20, 786–802 (2008).2. Treffon, P., Rossi, J., Gabellini, G., Trost, P., Zaffagnini, M., Vierling, E. Quantitative proteome profiling of a S‐nitrosoglutathione reductase (GSNOR) null mutant reveals a new class of enzymes involved in nitric oxide homeostasis in plants. Accepted. Front. Plant Sci. (2021).