bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of cinnamon metabolites on glucose utilization and glycolysis (1045.43)
Authors: Jiang T. Alan , Moss‐Pierce Tijuana , Tu Zheng 
Published: June 2021

Studies have shown that proanthocyanidins (oligomers/polymers of flavan‐3‐ols) in cinnamon may play a role in hypoglycemic activities. However, bioavailability of the oligomers and polymers is low, and they need to be biotransformed by the colonic microbiota into smaller metabolites in order to be absorbed. Therefore, phenolic metabolites, rather than the original high molecular weight compounds may be responsible for the biological activity derived from cinnamon consumption. We examined the effect of the following metabolites on glucose metabolism in vitro: 3 Hydroxyphenylacetic acid (3‐HPAA), 3,4‐dihydroxyphenylacetic acid (3,4‐DHPAA), 3‐(4‐hydroxyphenyl)propionic acid (3‐(4‐H)PA), 4‐hydroxyphenylacetic acid (4‐HPAA), α‐hydroxyhippuric acid, ferulic acid, and 4‐hydroxybenzoic acid. All were purchased from Sigma‐Aldrich. Glucose consumption in serum starved C2C12 myocytes culture medium was measured using a glucose assay kit (Sigma‐Aldrich). Glycolysis was measured in C2C12 myocytes by the extracellular acidification rate (ECAR) using a Seahorse XF96 analyzer with 10mM glucose. 3‐HPAA, 3,4 DHPAA, and ferulic acid increased glucose utilization by 60%, 118%, and 247%, respectively. In addition, 3,4‐DHPAA and ferulic acid increased glycolysis by 57%, and 62%, respectively. It therefore appears that certain proanthocyanidin metabolites exert biological activity in vitro.Grant Funding Source: This research was supported by McCormick and Company.
Modulation of estrogen metabolism by broccoli: mechanistic investigation of cancer prevention (1045.45)
Authors: Gaikwad Nilesh , Kucera Heidi , Olivera Deborah , Shibamoto Takayuki 
Published: June 2021

Modulation of Estrogen Metabolism by Broccoli: Mechanistic Investigation of Cancer Prevention Nilesh W. Gaikwad, Heidi R. Kucera, Deborah, Olivera, Taka Shibamoto Departments of Nutrition and Environmental Toxicology, University of California, Davis, CA 95616, USA Broccoli is commonly used as a popular vegetable in the human diet. Recently, broccoli sprouts have also received much attention because of their different bioactive constituents from mature broccoli. They are reported to be more effective in the prevention of carcinogenesis by reactive oxygen species than broccoli. However, very little information on their biological activities such as, antioxidant, anti‐inflammatory, anticancer, of broccoli sprouts, are available. Moreover, there are no reports on the antioxidant activity of the volatile compounds from broccoli sprouts. We hypothesize that bioactive compound(s) from broccoli sprouts could modulate estrogen metabolism towards non carcinogenic pathways resulting in a significant preventive effect. Present evidence shows the involvement of estrogens in carcinogenic processes within estrogen‐responsive tissues including the breast, endometrium and ovaries. Epidemiologic studies have indicated that breast cancer risk is higher in women with early menarche and late menopause, who have longer lifetime exposure to estrogens. Obese postmenopausal women have higher serum concentrations of free estrogen and are at greater risk of breast cancer. Gathering data suggests metabolism of estrogens to methides as well as catechol‐o‐quinones play a major role in estrogen carcinogenesis. To test our hypothesis, we have treated the MCF 10A cells with estrogens and broccoli sprout extract and using recently developed mass spectrometry based metabolomic platform we investigated the molecular mechanisms which leads to alteration of estrogen metabolic pathways.Grant Funding Source: NA
Versican is a critical extracellular matrix protein in lung and brain during embryonic development in the mouse (1046.9)
Authors: Snyder Jessica , Washington Ida , Chang Mary , Frevert Charles 
Published: June 2021

Versican, a chondroitin sulfate proteoglycan and a component of the extracellular matrix, is associated with fatal cardiac malformations if absent during embryonic development and is increased with injury in adult tissues. Changes in versican gene expression and accumulation have not been examined comprehensively in the lung and brain during mid to late gestation in the mouse. The objective of this work was to determine if versican accumulation and degradation change during lung and brain embryogenesis. Quantitative PCR and immunohistochemistry were used to measure versican expression, accumulation and degradation in the lung, brain, heart and liver of C57Bl/6 mice for five consecutive days during embryonic development. Consistent with previous reports, we found that total versican gene expression was higher in embryonic tissues when compared to adults and the highest total versican gene expression in embryos occurred at embryonic day 13.5 (E13.5). At E15.5, versican gene expression was lowest in the liver, highest in the brain, and intermediate in the heart and lung. There was differential expression of versican isoforms, with V0, V1, and V2 isoforms increased in the brain but only V0 and V1 isoforms increased in the lung. Versican expression in the lung peaked at E13.5, at which time there was strong versican immunoreactivity in the interstitium and airway basement membrane but little accumulation in airway epithelial cells. In the brain, immunoreactivity was strongest in the roof of the hind brain and neopallial cortex and in the deeper white matter tracts, and at later gestational ages there was little accumulation in the developing choroid plexus and periventricular gray matter. During embryonic development of the lung and brain, we found areas of versican expression that colocalized with the expression of DPEAAE, a versican degradation product. These results suggest that versican plays an important role in embryogenesis of the lung and brain.Grant Funding Source: Supported by HL098067, RR030249
Immunoexpression of the human kallikrein‐10 in surgically removed human pituitary tumors (1048.12)
Authors: Rotondo Fabio , Di Ieva Antonio , Kovacs Kalman , Syro Luis , Cusimano Michael , Yousef George 
Published: June 2021

Human kallikrein 10 (hK10), a serine protease, plays an important role in the regulation of cell proliferation and tumor growth. Its presence was demonstrated in lactotrophs and corticotrophs of the nontumorous human pituitary. In this work, we investigated hK10 immunoexpression in 59 surgically removed human pituitary tumors. Tissues were fixed in formalin, embedded in paraffin. Immunostaining was performed by the streptavidin‐biotin‐peroxidase complex protocol using the LSAB+ Kit (DAKO, Carpenteria, CA) and an hK10‐specific rabbit polyclonal antibody (1:150).Results showed that lactotroph adenomas removed from patients receiving no dopamine agonist medication were conclusively immunopositive for hK10. Immunopositivity was localized in the cytoplasm and was clearly visible in many adenoma cells. In the various tumor types (silent corticotroph subtype 1 & 2, oncocytic and gonadotrophic adenomas, somatotroph adenomas, and carcinomas), immunopositivity was very mild, seen only in few unevenly distributed tumor cells. Previous studies showed that hK10 possesses tumor suppressing properties and is decreased in rapidly growing tumor cells. Future studies should focus on hK10 immunoexpression in lactotroph adenomas exposed to dopamine agonist therapy which inhibits tumor growth.This study was supported by the Jarislowsky and Lloyd Carr‐Harris Foundations.
Ameloblastoma driver mutations revealed by next‐generation sequencing of formalin‐fixed paraffin‐embedded specimens (1048.18)
Authors: Pollack Jonathan , McClary Andrew , Sweeney Robert , Biscocho Jewison , Myers Benjamin , Neahring Lila , Kwei Kevin , Qu Kunbin , Gong Xu , Ng Tony , Jones Carol , Varma Sushama , Odegaard Justin , Rubin Brian , Troxell Megan , Pelham Robert , Zehnder James , Beachy Philip , West Robert 
Published: June 2021

Rare cancer types are not only understudied, but are typically represented by formalin‐fixed paraffin‐embedded (FFPE) (rather than freshly‐frozen) specimens that are suboptimal for genomic analysis. Ameloblastoma is one such rare tumor type, thought to arise from ameloblasts, the cells that deposit enamel during tooth development. Though typically benign, ameloblastomas are locally destructive to the jaw and face, and new non‐surgical interventions are needed. To discover novel driver mutations and therapeutic targets, we optimized methods and performed whole‐transcriptome sequencing and/or targeted exon sequencing (TruSeq Cancer Panel) of 8 FFPE cases. Identified mutations were verified, and then evaluated on a larger, independent set of 22 FFPE cases by PCR and Sanger sequencing. From the analysis, we identified recurrent somatic mutations in three key developmental or signaling pathways, including Hedgehog, fibroblast growth factor, and MAP kinase pathways. Functional interrogation of a novel Hedgehog pathway mutation confirmed increased basal pathway activity, and defined the response profile to various pharmacologic Hedgehog inhibitors. Together, our results define new ameloblastoma drivers and nominate new molecularly‐directed therapies for this rare but disfiguring disease. More generally, our findings validate a robust approach for discovering driver mutations in rare cancers.Grant Funding Source: Supported by Stanford University Department of Pathology
The effect of CBD, CBG and a combination of CBD plus CBG on Haccat human skin carcinoma cells (1048.19)
Authors: Javid Farideh , Afshinjavid Saeed 
Published: June 2021

The effect of CBD, CBG and a combination of CBD plus CBG on Haccat human skin carcinoma cells Cannabinoid pharmacology has received attention in cancer research for their potential in combating cancer (Alexander et al., 2009). The present study investigated the potential anti‐tumour activity of cannabidiol and cannabigerol on skin tumour cells.Haccat cells (American Type Culture Collection) were grown and maintained in DBEM medium plus10% fetal bovine serum at 37oC, 5% CO2 .The cells were plated in 96‐well culture plates at a density of 1x104 cells/well and allowed to adhere at 37oC for 24 hours. The following day, various doses of the compounds or were added to the cells and further incubated for 24, 48, 72 and 96 hours. Then the supernatant was removed and MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide) was added for 4 hours. The ability of cells to form formazan crystals by active mitochondrial respiration was determined after dissolving the crystals in DMSO. Cytotoxicity was expressed as a relative percentage of the absorbance measured at 540 nm.CBD induced dose‐dependent cytotoxic effect (IC50 of 4.83+1.4, 4.75+0.06; 5.22+0.3 and 5.19+1.27 micromolar for 24, 48, 72 and 96 h contact time respectively). Addition of CBG also induced cell toxicity with IC50 of 1.96+0.3, 5.5+0.34, 8.32+0.19 and 7.85+1.47 for 24, 48, 72 and 96 h respectively. A combination of CBD plus CBG induced cell toxicity with an IC50 of 2.4 + 0.05 over 96 h contact time.The data suggest a significant (p<0.001) cytotoxicity when compounds are applied in combination.We thank GW Pharmaceuticals for providing cannabinoid extracts.Grant Funding Source: University of Huddersfield
An upregulated long noncoding RNA RP3‐337D23.3 in lung adenocarcinoma in never‐smokers promotes metastasis (1049.1)
Authors: Yin Rong , Qiu Mantang , Jiang Feng , Xu Lin 
Published: June 2021

Objective: The present study was designed to screen and validate lung adenocarcinoma in never smokers (LACINS) specific long noncoding RNAs (lncRNAs).Methods: A custom‐designed human lncRNA expression microarray was utilized to determine lncRNA expression profiles of LACINS and indentify LACINS specific lncRNA. Five LACINS and paired adjacent normal lung tissues were used for microarray analysis and another 10 LACINS were used for further searching of LACINS specific lncRNAs by quantitative real‐time polymerase chain reaction. Function of RP3‐337D23.3 was investigated by cell proliferation, transwell, and wound healing assay in vitro.Results: A total of 3353 lncRNAs were differentially expressed in LACINS (fold change >2), 1509 upregulated and 1849 downregulated. Eleven candidate LACINS specific lncRNAs were identified and further validated in 10 pairs of LACINS and corresponding lung tissues. We identified RP3‐337D23.3 as a LACINS specific lncRNAs which is significantly up‐regulated in tumors as compared with adjacent normal tissues. Additionally, RP3‐337D23.3 is significantly overexpressed in LACINS with metastasis compared with those without metastasis (p=0.0006). In vitro siRNA mediated silencing of RP3‐337D23.3 led to remarkable decreased invasion, migration, and proliferation abilities of H1975 cell line.Conclusion: We identified the first LACINS specific lncRNA, RP3‐337D23.3, and provided preliminary evidence that RP3‐337D23.3 could promote LACINS metastasis.Grant Funding Source: National Natural Science Foundation of China (81372321)
Glypican‐5 regulates lung cancer cell metastasis through Wnt signaling pathway (1049.3)
Authors: Xu Lin , Yin Rong , Jiang Feng , Yang Xin 
Published: June 2021

Glypican‐5 (GPC5) may be a potential tumor suppressor gene in non‐small cell lung cancer (NSCLC). Previously, we determined that GPC5 over‐expression in lung cancer cells could significantly suppress their migration, invasion, and proliferation activities and also induced G1/S phase arrest of the cells in vitro. However, the signaling mechanism of GPC5 is unclear. We reported here that GPC5 inhibited the metastasis of lung cancer cells in vitro and in vivo by blocking canonical WNT signals. Wnt signaling in vitro and in vivo was assessed in A549 cells transfected with GPC5 recombinant lentivirus. The interaction between GPC5, E‐Cadherin andβ‐Catenin was assessed by coimmunoprecipitation. β‐catenin‐dependent transcriptional activity was assessed by the TOPFLASH luciferase assay. Expression of GPC5, Wnt signaling molecules and target genes was analyzed by real‐time PCR and Western blot. We found that GPC5 is able to inhibit canonical Wnt signals involved in cell migration, invasion, and proliferation. Our results are consistent with the potential role of GPC5 as a metastasis suppressor in lung cancer.Grant Funding Source: National Natural Science Foundation of China (81201830)
Evaluation of tissue section cryostorage on immunohistochemistry (1050.1)
Authors: Estrada Larissa , Amaral Ana , Giannaris Eustathia , Heyworth Nadine , Mortazavi Farzad , Ngwenya Laura , Roberts Debra , Robinson Amy , Killiany Ronald , Rosene Douglas 
Published: June 2021

Storage of tissue sections for significant periods is often required to allow tissue from multiple samples, acquired over months to years, to be processed together, in the same reagents, for quantitative studies using immunohistochemistry. Several protocols for storage in sucrose with different additives have been reported that assert there is no effect on immunogenicity. However, there have been no quantitative assessments of how long‐term frozen storage in any cryoprotectant affects immunogenicity. The present study analyzed existing datasets, derived from monkey brain sections stored at ‐80°C in 15% phosphate buffered glycerol to determine whether time frozen in this cryoprotectant affected stereology counts of cells immunostained for NeuN, Parvalbumin, Orexin‐A, Doublecortin, BrdU or pro‐BDNF. Multiple regression analyses controlling for age and sex of the subjects in the different data sets showed that time in 15% glycerol did not affect stereological counts for any marker. Experiments are underway to compare our 15% glycerol cryoprotectant to four 30% sucrose based cryoprotectants for preservation of immunogenicity after frozen storage.Grant Funding Source: Supported by NIH grants R01‐AG043640 and R01‐AG042512
Laminitis‐specific microRNAs in equine plasma (1050.6)
Authors: Kim Myung‐Chul , Lee Seung‐Woo , Cui Feng‐Ji , Ryu Doug‐Young , Kim Yongbaek 
Published: June 2021

Purpose: To identify laminitis‐specific plasma microRNAs in a horse Method and materials: In order to induce laminitis experimentally, oligofructose was administered to a horse through nasogastric tube. Peripheral blood was collected from the jugular vein at the time of pre/post induction of laminitis. After euthanasia, various tissues including laminae were collected from the horse. Total RNA was extracted and subjected to the reverse transcription to cDNA. The cDNA products of the small RNA fragments ranging from 40bp to 60bp were sequenced directly using Illumina HiSeq 2000 sequencer.Results: A total of 288 miRNAs were identified in laminar tissues and 262 miRNAs were identified in the plasma of horse with laminitis. Fifteen miRNAs were identified as laminitis‐specific plasma miRNAs, based on the following criteria: laminar tissue‐specific expression compared to other tissues and significant increase of the plasma miRNA level after the induction of laminitis.Conclusions: The present study revealed a subset of plasma miRNAs specific to horse laminitis, suggesting the potential value of the miRNAs as a biomarker for the diagnosis of equine laminitis.Grant Funding Source: Supported by Bio‐industry Technology Development Program of iPET, Ministry of Agriculture, Korea
Low molecular weight fucoidan against diabetic nephropathy (1051.17)
Authors: Zhou Hong 
Published: June 2021

The objective of the present study was to investigate the therapeutic effect and underlying mechanisms of LMWF on diabetic nephropathy.LMWF was administered to the diabetic rats at a daily dose of 100 mg/kg body weight by intragastric administration. Following 12 weeks treatment, we measured body and kidney weight, blood glucose, blood pressure, creatinine and urea concentration in serum and urine, urine output, urinary osmolality, urinary protein excretion. α‐SMA, E‐cadherin, fibronectin, TGF‐β1, CTGF were measured by Western blot to evaluate the level of epithelial‐to‐mesenchymal transition (EMT). In addition, the phosphorylation of Akt, Smad3, p38 and ERK in renal tissue were evaluated. In virio, HK2 renal tubular epithelial cells pretreated with LMWF were stimulated by the recombinant human TGF‐β1 (10 ng/ml) for 48 h. α‐SMA, vimentin, E‐cadherin, fibronectin, CTGF were measured by Western blot to evaluate the level of EMT. In addition, activity of Akt, Smad and MAPK signal pathway in HK2 cell were evaluated.Renal expression of TGF‐β1, CTGF, fibronectin and α‐SMA protein were increased significantly in diabetic rats compared to those in the controls. E‐cadherin protein expression in diabetic kidneys decreased significantly. LMWF effectively reduced hypertension, proteinuria, renal TGF‐β1, CTGF, fibronectin and α‐SMA up‐regulation, as well as renal E‐cadherin down‐regulation in diabetic rats without a significant effect on blood glucose. LMWF also inhibited the activation of Smad, Akt, MAPK pathways. In vitro, LMWF alleviated TGF‐β1 induced EMT in HK2 cells via inhibition of Smad, Akt, MAPK pathways in a dose‐dependent manner.Our in vivo and in vitro studies show that LMWF ameliorates diabetic nephropathy, possibly through inhibiting the expression of TGF‐β1 and Smad, Akt, MAPK signaling pathways downstream, then subsequent suppressing EMT. The data provide evidence that LMWF may serve as a potential therapeutic agent for diabetic nephropathy.
microRNA‐34a mediates impaired angiogenesis in diabetes: role of SIRT1 and metformin (1051.2)
Authors: Arunachalam Gnanapragasam , Samuel Samson , Ghosh Suparna , Triggle Christopher , Ding Hong 
Published: June 2021

Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in patients with diabetes mellitus. In particularly, hyperglycemia‐associated impaired angiogenesis and vascular dysfunction critically contributes to the pathogenesis of diabtes‐associated CVD. Recent findings demonstrated the importance of microRNAs (miRNAs) in the vasculature and emerged as important regulators of angiogenesis. Our data with microvascular endothelial cells (MMECs) cultured in high glucose (HG, 40 mM) indicate a significant increase in miR‐34a expression and this induction paralleled and correlated with altered expression of SIRT1, eNOS (phospho/acetylation) and makers of angiogenesis (VEGF, angiopoietins and thrombospondins) as well as a significant impairment of tube‐forming activity (angiogenesis) when compared with normal glucose (NG, 11mM). Interestingly, inhibition of miR‐34a (anti‐miR) increased SIRT1 expression and attenuated changes in downstream signaling and impaired angiogenesis in HG‐exposed MMECs. Conversely, treatment with metformin (50μM) also inhibited miR‐34a expression and attenuated HG‐induced impaired angiogenesis in MMECs. miR‐34a, via the regulation of SIRT1 expression, has an anti‐angiogenic action in microvascular endothelial cells that is attenuated by metformin. miR‐34a may represent a new therapeutic target for the prevention/treatment of diabetic vascular disease.Grant Funding Source: This project was funded by grant # UREP 13‐116‐3‐024, NPRP 5‐149‐3‐040
Sodium 4‐phenylbutyrate reverses glucose deprivation‐induced endoplasmic reticulum stress and death signal in cultured mouse microvascular endothelial cells (1051.3)
Authors: Samuel Samson , Arunachalam Gnanapragasam , Ghosh Suparna , Emara Mohamed , Ding Hong , Triggle Chris 
Published: June 2021

In diabetes mellitus, despite higher than normal levels of blood glucose, deficiencies in signaling mechanisms results in a failure to utilize glucose, creating a scenario of variable glucose deprivation (GD). Reports suggest that stringent control of blood glucose in diabetic subjects invariably increases the risk of hypoglycemia, the severity of which is associated with increased cardiovascular risk. However, in such conditions the possible role of endoplasmic reticulum (ER) stress associated cell death during GD in microvascular endothelial cells has not been studied. Mouse Microvascular Endothelial cells (MMECs) subjected to GD for 24h showed significant increase in the levels of GRP78, p‐eIF2α, ATF4 &amp; pro‐apoptotic CHOP, while levels of total PERK decreased &amp; total‐ eIF2α did not change. Autophagic markers showed significant up‐regulation upon GD. 10mM of 4PBA, a chemical chaperone, significantly reversed the GD induced changes in ER stress &amp; autophagic markers. Immunocytochemical analysis revealed that stress granules (SGs) are assembled at early hours (1h‐8h) of GD while very few SGs are observed at 16h‐24h. Our data suggests that GD induced ER stress &amp; associated activation of death signal could play a major role in GD associated endothelial cell loss, which could be reversed by treatment with suitable chaperones.Grant Funding Source: Supported by: QNRF grants # NPRP 4‐910‐3‐244 & JSREP 3‐016‐3‐009