bluuebunny/crossref_metadata_2025_split huggingface dataset:
Potential Role For A Phosphoserine Aminotransferase 1 And Pyruvate Kinase M2 (PSAT1:PKM2) Functional Interaction In Lung Cancer Cells
Authors: Clem Brian F , Kruer Traci , Bradley James , Merchant Michael , Trent John O , Sit Rumeysa Biyik 
Published: November 2023

Cellular production of serine provides precursors for numerous anabolic reactions, particularly for proteins and nucleic acid synthesis and for one carbon metabolism, and this pathway has been demonstrated to be elevated in many different cancer types. In addition, metabolites within serine synthesis can stimulate other processes required for neoplastic growth, including glucose metabolism, specifically through the activation of the glycolytic enzyme pyruvate kinase M2 (PKM2). However, the complete mechanism by which serine biosynthesis facilitates metabolic or cellular changes necessary for tumor growth are unclear and is further confounded by the observation that suppression of serine biosynthetic enzymes disrupts tumorigenesis even in the presence of exogenous serine. Phosphoserine aminotransferase 1 (PSAT1), the second enzyme in serine synthesis, catalyzes the conversion of 3‐phosphohydroxypyruvate to phosphoserine and increased expression correlates with poorer overall survival in lung cancer patients. In order to determine whether PSAT1 may contribute to lung cancer progression, PSAT1 expression was stably silenced in A549 and H358 lung cancer cells. Although a modest change in cell proliferation was observed upon PSAT1 knock‐down compared to control, there was a robust decrease in soft‐agar colony formation in both cell types. PSAT1 suppression was accompanied by distinct morphological changes and an increase in E‐cadherin expression, suggesting a potential switch in the epithelial‐mesenchymal transition (EMT), which underlies the metastatic potential of cancer cells. Accordingly, PSAT1 suppression resulted in a decrease in cellular motility as demonstrated through wound healing assays. To ascertain whether specific PSAT1 mediated protein:protein interactions may contribute to the pro‐tumorigenic potential of serine metabolism, we performed GST‐PSAT1 pull‐down assays that revealed a potential direct association with PKM2, which was confirmed by co‐immunoprecipitation. In vitro recombinant enzyme assays found that PSAT1 activates PKM2, but not PKM1 and, addition of serine or fructose‐1,6‐bisphosphate further activated PKM2, suggesting that PSAT1‐dependent stimulation is distinct from other allosteric modulators. Lastly, based on in silico modeling, site‐directed mutagenesis of PKM2 identified amino acid residues required for this protein interaction. Taken together, elucidating the functional role of PSAT1‐PKM2 in lung cancer cells may yield valuable information about the relationship between serine‐glucose metabolism and EMT transition in cancer cells and provide an additional therapeutic target for cancer treatment.Support or Funding InformationRSG 13‐139‐01(American Cancer Society) and CA166327 (NCI) to BFC
Isolation, Identification, and Characterization of Local Wild Yeasts for Use In Fermentation
Authors: DeLong Lauren Nicole , Noone Brenna , Erickson Les 
Published: November 2023

Inspired by spontaneously fermented beers, such as Belgian Lambic beers, American breweries have moved beyond using exclusively domesticated strains of the beer‐making yeast, Saccharomyces cerevisiae, to experimenting with a variety of different yeast species to produce unique flavors and aromas. The so‐called “funky,” “wild,” and “sour,” beers are produced using uncommon microorganisms either during or after the primary ferment. The goal of our research is to isolate, identify, and characterize wild yeasts from the local environment and to make them available to both commercial and craft brewers for making novel beers. The use of locally‐sourced yeasts could also be used by commercial brewers as a marketing angle, as is often done with local hops, grains, or water. As a source of wild yeasts, we obtained various wild and cultivated fruits growing in rural Maryland and a sample of Belgium‐style ale being aged in used oak wine barrels at Evolution Craft Brewing Company (Salisbury, MD). Yeast colonies were isolated using yeast extract/peptone/dextrose (YPD) agar plates containing antibiotics and identified by colony morphology as well as PCR amplification and sequencing of the 5.8S rRNA genes. The identified yeasts belong to Saccharomyces, Pichia, Candida, Fusarium, Torulaspora, and, most interestingly, Brettanomyces genera. The Brettanomyces yeast, which was isolated from the wine barrel ale, is normally considered a wine spoilage yeast but increasingly is being employed to create “funky” and sour flavors in beer. The growth and fermentation abilities of eight different yeast isolates were analyzed for growth in different liquid medias (YPD, hopped barley‐malt extract, and merlot grape juice) using 96‐well plates and spectroscopy. Growth rates were calculated using a modified grofit package in RStudio. Ethanol production will be determined using a colorimetric assay. Thus far, the most compelling discovery is a wild strain of Saccharomyces cerevisiae that grows comparable to a commercially‐available brewing yeast and produces pleasant, fruity aromas when fermenting barley malt extract. The complete characterization of these wild yeasts will be reported in order to identify novel wild yeasts that can be domesticated for local craft brewers.Support or Funding InformationSpecial acknowledgments to the Salisbury University Department of Biological Sciences, the Henson Undergraduate Research Award, Evolution Craft Brewery, and the National Science Foundation for funding and equipment, as well as Dr. Philip D. Anderson, Rachel Keuls, and Brittney Lozzi for their support.
Novel Crosstalk between Insulin and TGF‐beta signaling in Vascular Endothelial Cells
Authors: Shah Nirav , Pan Christopher , Kumar Sanjay , Zaman Naveed , Elmasry Fatema , Lee Nam Y 
Published: November 2023

Angiogenesis is the formation of new blood vessels from preexisting vasculature, an essential process during development and tumor growth. As a TGF‐β receptor complex primarily expressed in proliferating endothelial cells (ECs), endoglin and ALK1 activate the canonical Smad1/5/8 signaling to promote normal and tumor‐associated angiogenesis. Here we report an unexpected crosstalk between insulin and endoglin/ALK1‐dependent TGF‐b signaling pathways. We found that insulin rapidly activates the canonical TGF‐β and BMP‐9‐induced Smad1/5/8 pathway in numerous EC types tested. Co‐immunoprecipitation, immunofluorescence and targeted knockdown studies reveal that insulin activates Smad1/5/8 through a novel trimeric interaction comprising the insulin receptor, endoglin and ALK1. Importantly, our PCR gene profiling results indicate that insulin‐induced Smad1/5/8 signaling elicits a transcriptional response quite different than those normally activated by TGF‐b or BMP‐9. Given that type 2 diabetic patients are highly susceptible to a number of vascular‐related diseases including proliferative retinopathy, our results strongly implicate aberrant Smad1/5/8 signaling with hyperinsulinemia and insulin resistance during pre‐diabetic and diabetic disease progression.Support or Funding InformationRO1 NCI
Alcohol and HIV Protease Inhibitor‐induced inflammasome activation and hepatic lipotoxicity
Authors: Hinton Michael , Pandak William , Hylemon Phillip , Zhou Huiping 
Published: November 2023

The development of highly‐active‐antiretroviral therapy (HAART) has allowed management of HIV and extended the lives of those infected. Alcohol abuse, which is very common in HIV‐1 infected patients, is one of the most important co‐morbid risk factors for liver injury and has been associated with the occurrence of serious metabolic syndrome and subsequent discontinuation of HAART in HIV patients. The importance of the NLRP3 inflammasome in liver diseases has been recognized. Kupffer cells (KC) are the major resident macrophages and play a critical role in regulating hepatic inflammatory response. We have shown that HIV protease inhibitors (PI) increase the production of mature IL‐1β production in macrophages suggesting the activation of NLRP3. The aim of this study is to examine the effect of alcohol and HIV PIs on NLRP3 activation in KCs and characterize the impact of inflammasome activation in KCs on alcohol and HIV PI‐induced hepatic lipotoxicity.MethodsPrimary mouse KCs were isolated from rat and treated with individual HIV PIs with or without alcohol. The mRNA and protein levels of inflammasome components including NLRP3, ASC, and pro‐caspase were measured by real‐time RT‐PCR and Western blot analysis, respectively. The protein levels of IL‐1β and IL‐18 in cell culture medium were measured using ELISA analysis.ResultsWe found both alcohol and HIV PI‐induced activation of NLRP3 in KCs. The expression of mature IL‐1β and IL‐18 were increased by alcohol and HIV PIs. The active form of caspase‐1 was also increased.ConclusionsActivation of inflammasome in KCs is a key cellular mechanism underlying alcohol and HIV PI‐induced hepatotoxicity.Support or Funding InformationNational Institutes of Health (R21 AI068432, R01 AT04148, R01 AI057189, P01 DK38030, P30 CA16059), VA Merit Award, Merck research fund and Jeffress GrantNational Institutes of Health (F31 AA024713‐01)
Inflammatory generation and signaling actions of conjugated nitro‐linoleic acid
Authors: Schopfer Francisco Jose , Vitturi Dario , Minarrieta Lucía , Salvatore Sonia Rosana , Khoo Nick , Jobaggy Soma , Li Lihua , Woodcock Steven , Berman Rebecca , Ferreira Ana , Freeman Bruce , Villacorta Luis 
Published: November 2023

Inflammatory responses result in the expression and activation of pro‐oxidant enzymes such as the inducible isoform of nitric oxide synthase (iNOS) and NAPDH oxidases, leading to the generation of the nitrating species nitrogen dioxide (•NO2). Conjugated linoleic acid is a primary substrate for biological lipid nitration in the gastric compartment giving rise to the formation of conjugated nitro‐linoleic acid (NO2‐CLA). NO2‐CLA exhibit reversible electrophilic reactivity and interact covalently with nucleophilic cysteines in proteins and low molecular weight compounds. This reactivity is pivotal for their biological actions including Nrf2 induction, partial PPARγ agonism, heat shock response activation and TLR4/NF‐κB/STAT1 inhibition. Herein we show that macrophage activation leads to NO2‐CLA formation both in vitro and in vivo, as characterized by high resolution mass spectrometry and quantified using isotopic dilution HPLC‐MSMS. CLA nitration by activated RAW264.7 cells results not only in the dose‐ and time‐dependent formation of the parent NO2‐CLA compound but also in the generation of secondary electrophilic and non‐electrophilic b‐oxidation metabolites. The formation of two positional isomers 9‐NO2‐CLA and 12‐NO2‐CLA with different metabolization rates but similar signaling potency was also observed. Addition of either mixed or pure NO2‐CLA positional isomers to cells in culture led to antioxidant response element (ARE) activation and to dose‐dependent expression of Nrf2‐regulated genes. Similarly, NO2‐CLA attenuated TNF‐a mediated NF‐kB response element activation and decreased iNOS expression and pro‐inflammatory cytokine release in LPS/IFN‐g activated RAW264.7 cells. To assess the physiological relevance of these observations, a mouse zymosan A peritonitis model that promotes CLA nitration was utilized. Animals were administered vehicle, or synthetic NO2‐CLA and then inflammatory cell recruitment and clearance from the peritoneal cavity was monitored. NO2‐CLA administration led to decreased recruitment as assessed by flow cytometry. Our work demonstrates that NO2‐CLA generation might constitute an endogenous adaptive mechanism capable of modulating the severity of the inflammatory response by favoring resolution and inducing the expression of both cytoprotective and antioxidant genes.Support or Funding InformationThis study was supported by NIH grants R01‐HL123333 (LV), R01‐AT006822 (FJS) and K01‐HL133331 (DAV), and AHA #14GRNT20170024 (FJS).
Sterol O‐acyltransferase 1 enhances cholesterol esterification via cyclic AMP‐dependent pathway in the yolk sac membrane endodermal epithelial cells
Authors: Lin Han‐Jen , Wang Siou‐Huei , Chen Yu‐Jen , Mersmann Harry John , Ding Shih‐Torng 
Published: November 2023

During avian embryonic development, absorptions of nutrients is mainly responsible by endodermal epithelial cells (EECs) in the yolk sac membrane (YSM). Yolk lipids are actively absorbed in late stages of incubation. Sterol O‐acyltransferase (SOAT), regulating the cholesterol esterification with long‐chain fatty acid, is the key for lipid utilization during embryonic development. Expressions of SOAT1 are higher than SOAT2 in YSM during embryonic development of Japanese quail, we hypothesized that SOAT1 is greatly involved to mediate uptake of free fatty acid and yolk cholesterol into EECs, and also responsible for absorption and transportation of lipids into developing embryos. Treatments of glucagon, isobutylmethylxanthine (IBMX), a phosphodiesterase inhibitor for increasing cyclic AMP (cAMP) concentration, and low glucose increased SOAT1 mRNA expression in EECs and hepatocytes, suggesting that SOAT1 is regulated by a cAMP‐dependent pathway. Expression of SOAT1 was increased by adenylate cyclase activator (forskolin) and cAMP analog (dibutyryl‐cAMP) treatments in EECs and hepatocytes. Furthermore, the protein kinase A (PKA) activity was increased by IBMX treatment, whereas co‐treatment of PKA inhibitor, H89, negated the increase in PKA activity. Cyclic AMP‐induced EECs had higher activity of cholesterol esterification and thereby more intracellular lipids than untreated EECs. In conclusion, expressions of SOAT1 is regulated via cAMP‐dependent pathway, therefore, factors that regulate PKA will increase the expression of SOAT1 to improve the utilization of lipids in the EECs and modify embryonic growth.Support or Funding InformationSterol O‐acyltransferase 1, cholesteryl ester, cAMP‐dependent pathway, yolk sac membrane, avian embryos
Glycolipid Storage and Phenotypes in a New Rat Model of Fabry Disease
Authors: Miller James J , Aoki Kazuhiro , Murphy Carly A , Stucky Cheryl L , Kassem Iris S , Tiemeyer Michael , Dahms Nancy M 
Published: November 2023

Fabry disease is an X‐linked lysosomal storage disease caused by mutations in the GLA gene, which encodes for the acid hydrolase, α‐galactosidase A (α‐Gal A). When α‐Gal A activity is deficient, glycolipids with terminal α‐galactosyl residues accumulate and lead to cellular dysfunction and ultimately organ pathophysiology. Patients with Fabry disease experience severe pain starting in childhood, develop corneal and lenticular opacities, and eventually suffer from end stage renal disease. In order to study the development of these phenotypes on a molecular level, we generated an α‐Gal A deficient (Gla KO) rat using CRISPR/Cas9 technology. Gla KO rats have undetectable serum α‐Gal A activity, but have similar levels of serum α‐mannosidase activity as wild type (WT) rats. As measured by mass spectrometry in serum and tissues (kidney and heart), Gla KO rats have dramatic increases in molecules containing terminal α‐galactosyl residues, including globotriaosylceramide (>100‐fold), globotriaosylsphingosine (>30‐fold), and blood group B glycolipids (>10‐fold), which are established biomarkers for patients with Fabry disease. Non‐α‐galactosyl glycolipids, such as ganglioside GM3, exist in similar concentrations in both Gla KO and WT rat serum and tissues. Electron microscopy of Gla KO kidneys and dorsal root ganglia reveal the presence of electron dense lamellar structures, which are classically seen in patients with Fabry disease. On a macroscopic level, male Gla KO rats weigh ~10% less on average and have enlarged tongues (P < 0.001) compared to WT controls. Behavioral studies show that Gla KO rats have increased sensitivity to touch and sharp punctate force (P < 0.001), which are suggestive of a pain phenotype. Slit lamp biomicroscopy shows that male and female Gla KO rats develop corneal and lenticular opacities to a greater degree than WT controls (P < 0.01) by 13 weeks of age. Urinalysis shows that aging Gla KO rats develop increased urine calcium excretion (P < 0.001), increased urine flow (P < 0.001), and decreased urine osmolarity (P = 0.09), all of which suggest the development of distal tubule pathology. As the Gla KO rat recapitulates phenotypes seen in patients with Fabry disease, this rat model can be used to test new therapies and study the mechanistic basis of disease pathogenesis.Support or Funding InformationNIH funding: F30DK113641, K08EY024645, P30EY001931, R21NS095627, R24HL114474, T32GM080202
Serum Stable Isotope Biomarkers are Moderately Associated with Added Sugar, Fish and Meat Intake in 153 Postmenopausal Women
Authors: Yun Hee Young , Lampe Johanna W , Neuhouser Marian L , Tinker Lesley F , Prentice Ross L , O'Brien Diane M 
Published: November 2023

Stable isotope ratios are emerging dietary biomarkers that have not been extensively evaluated in the context of a typical US dietary pattern. Stable isotope ratios vary in the US diet; for example, the carbon isotope ratio (CIR) is elevated both in added sugars and meats, the nitrogen isotope ratio (NIR) is elevated in meats and fish, and the sulfur isotope ratio (SIR) is elevated in fish. In this study we tested whether serum CIR, NIR or SIR could predict intakes of added sugar, meats or fish intake, either alone or in combination. We also evaluated whether the CIR of serum amino acids showed more specific dietary associations. The study population was 153 postmenopausal women in Seattle WA who were participants in the Women's Health Initiative (WHI) Nutrition and Physical Activity Assessment Feeding Study (NPAAS‐FS). For a two week period each participant consumed a controlled diet that was designed to match her usual intake, and blood samples were collected before and after the period of dietary control. Post‐feeding serum stable isotope ratios were measured using continuous flow isotope ratio mass spectrometry and compared to participants' known dietary intake over the two week period using Pearson's correlation. Specifically, we examined intakes of animal protein, meat, poultry, fish, eggs, dairy, added sugar, and corn‐based foods. Animal protein intake (g/d) was associated with both the CIR (0.34) and the NIR (0.39). Among sources of animal protein, meat intake (g/d) was associated with the CIR (r = 0.32), and fish intake (g/d) was associated with the NIR (r = 0.57) and only weakly with the SIR (r = 0.18). None of these associations improved when biomarkers were used in combination. Added sugar intake (g/d) was associated with the CIR of serum alanine (r = 0.31), and meat intake (g/d) was associated with the CIR of serum essential amino acids (valine, leucine, isoleucine, and phenylalanine, r = 0.28 – 0.35). The findings that (1) added sugar intake was associated with serum alanine CIR but not total serum CIR and (2) fish intake was associated with serum NIR are consistent with previous results from an Alaska Native population. In addition, we found that meat intake was associated with total serum CIR as well as serum essential amino acid CIR. These data contribute to ongoing efforts to evaluate these emerging biomarkers of long‐term dietary intake in US populations.Support or Funding InformationThis project was supported by the National Institutes of Health National Cancer Institute: 5R21CA182674, 5R01CA119171.
Vitamin C content in dried and brewed green tea from the US retail market
Authors: Somanchi Meena , Phillips Katherine , Haile Ermias , Pehrsson Pamela 
Published: November 2023

BackgroundTea is the most consumed beverage after water in the world. Of the three main types of teas, namely black, oolong and green tea, the latter has been associated with a variety of health benefits, including cancer prevention, cardiovascular health, oral health and others, attributed to antioxidants activity as well as specific phytochemicals and nutrients it contains. Vitamin C is an essential nutrient, with a recommended daily intake of 60 mg, and higher intakes have been associated with numerous health benefits. Green and black tea are made from the leaves of Camellia Sinensis but green tea is less processed than black tea. Vitamin C concentrations exceeding 250 mg/100g have been reported in the unbrewed leaves, based on scientific literature for mostly Japanese green tea. Different cultivars and processing and brewing methods could significantly impact vitamin C in tea as consumed, and data on vitamin C in green tea available in the U.S. retail market are lacking. The goal of this study was to assess the vitamin C content of retail green tea bags in the U.S. retails market.MethodsSampling was done according to the National Food and Nutrient Analysis Program (NFNAP), which identifies top consumed products and implements a nationwide sampling based on market share. Samples of three brands were collected from each of the 12 locations. Samples from these 12 locations were composited randomly into pairs giving rise to six analytical samples. Six selected local samples of green tea bags were steeped in 230 ml of distilled hot water at 70°C, for two and four minutes and analyzed along with the corresponding unbrewed leaves. Vitamin C was determined as total ascorbic acid and dehydroascorbic acid, HPLC with UV detection. Certified reference materials and in‐house prepared control materials were included in each analytical batch.ResultsThe brewed tea contained <0.5 mg of vitamin C/100g, except for one local sample that contained 1 mg/100g; brewing time did not affect the results. Vitamin C in the unbrewed leaves ranged from < 3 to 178 mg/100g (mean 45.5; median 28.1).ConclusionAlthough this study did not screen all brands of green tea or possible brewing methods, the consistently negligible vitamin C in brewed tea suggests that the high vitamin C content in tea leaves should not be considered to transfer into the beverage as typically consumed in the U.S.