bluuebunny/crossref_metadata_2025_split huggingface dataset:
JAM‐A regulates the progression of Non‐alcoholic Fatty Liver Disease (NAFLD)
Authors: Kissig Megan Elizabeth , Naik Ulhas P. 
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD) is characterized by an abnormal amount of fat accumulation in the liver, specifically more than 5% fat by weight. Little is known about how the fat accumulates in the liver, but it has been found that intestinal permeability due to leaky tight junctions may be a contributing factor. Our lab studies junctional adhesion molecule‐A (JAM‐A), a protein located at the tight junctions of epithelial and endothelial cells. Through its ability to homodimerize at the apical part of the lateral membrane, JAM‐A helps regulate the permeability and stability of the junction.This study's aim was to find what effect JAM‐A has on the development of NAFLD. To analyze this relationship, groups of Jam‐A (+/+) and Jam‐A (−/−) mice were put on either a high‐fat or low‐fat diet for 20 weeks. During this time the mice were weighed every two weeks and blood samples were taken every four weeks. At the end of the 20 weeks, the mice were sacrificed and the livers and fat pads were removed.We found that the variation in weight between the Jam‐A (−/−) groups was greater than that between the Jam‐A (+/+) groups. Also, the average area of the adipocytes in the high‐fat Jam‐A (−/−) group was greater than the average area in the high‐fat Jam‐A (+/+) group. Since the liver sends fat to tissues in the form of cholesterol, we compared the levels in the plasma. The high‐fat Jam‐A (−/−) mice had significantly higher LDL‐cholesterol and total cholesterol levels in the plasma than the other groups as well. After examining the stained sections of the livers, it was found that the livers of the high‐fat Jam‐A (−/−) mice showed more fat droplet accumulation than the high‐fat Jam‐A (+/+) mice. This suggests that ultimately the absence of JAM‐A from the tight junctions promotes the development of NAFLD. This project was funded by the Arnold and Mabel Beckman Foundation.
Developmental regulation of caspases during death and regeneration events in an invertebrate chordate
Authors: Robison Leah Danielle , Lauzon Robert 
Published: June 2021

Programmed cell death (PCD) by apoptosis is an essential process in the development and maintenance of multicellular animals. We study the colonial marine urochordate Botryllus schlosseri which undergoes synchronized apoptosis throughout its entire adult generation every week. During a colony's death phase, the old adult cells are engulfed and recycled by circulating phagocytes simultaneously as asexually‐derived buds reach functional maturity. Thus, Botryllus is a unique model organism to study the cyclical regulation and function of cysteinyl aspartases called “caspases” during organismal apoptosis. It is well documented that caspases ‐8 and ‐9 play a role in apoptosis initiation, while caspases ‐3 and ‐7 are used in its execution. We have investigated whether caspase expression is developmentally regulated during asexual development in Botryllus. An expressed sequence tag (EST) sequencing project has uncovered several candidate sequences of the apoptotic death program, including several caspase orthologs. Using standard, semi‐quantitative PCR, we have previously demonstrated that initiator and executioner caspases are developmentally regulated in Botryllus, and exhibit maximal expression at the onset of a colony's death phase. However, as these experiments made use of genotypically distinct colonies for comparison, the effects of genetic background on caspase expression could not be excluded. In the current study, we sought to investigate the regulation of caspase expression using clonal replicates and quantitative polymerase chain reaction (Q‐PCR). Experiments are currently underway to determine whether caspase expression is associated with specific death and regeneration events in the lifecycle of Botryllus.Source of research support: Union College Internal Education Fund.
Insertion/Deletion Polymorphism of the Angiotensin Converting Enzyme Gene and Breast Cancer Risk
Authors: Javaid Naila Khatija , Fathima Husna , Abdulla Gulnaz , Al‐Ammari Abeer Saleh , Abbasi Fatma Ahmad 
Published: June 2021

BackgroundRenin‐angiotensin converting enzyme (ACE) polymorphism has recently been linked to the pathogenesis and prognosis of human cancers. This study aims to evaluate the association of ACE insertion or deletion (I/D) polymorphism with the risk of breast cancer incidence and the effect of genotype prognostic criteria of the tumor.MethodThe study included 36 breast cancer female patients and 46 healthy control females. DNA was extracted from tissue and blood samples, and polymorphism was detected by polymerase chain reaction followed by gel electrophoresis.ResultsThe distribution of the ACE genotype in the malignant group was 55.6% DD, 44.4% ID and 0% II. The DD genotype showed a significantly higher risk for the incidence of breast cancer (OR=3.9, 95% CI=1.5–10.2, p<0.05). Moreover, there was an association between the DD genotype and the risk for tumor metastasis (OR=1.7, 95% CI=0.3–11), poor differentiation (OR=1.5, 95% CI=0.4–5.8) and larger tumor size (OR=1.3, 95% CI=0.3–5.6).ConclusionOur findings suggest a probable prognostic value of the DD genotype in breast carcinoma.Research Supporter: Dubai Medical College for Girls, Dubai
Pokeweed antiviral protein (PAP) exhibits variable binding affinity to uncapped mRNA with differing structural elements
Authors: Friedland Diana E , DeGrazia Jeannine , Toney Alexandra 
Published: June 2021

Ribosome‐inactivating proteins (RIPs) exhibit N‐glycosidase activity by depurinating a surface exposed adenine of the α‐sarcin/ricin loop (SRL) of the large rRNA, ultimately causing cell death. They are found in numerous plant, fungi, and bacterial species and are powerful antiviral agents. PAP is a single‐chain RIP and also possesses mRNA recognition, cap binding, and depurination activities that are novel to RIPs. PAP's recognition of certain secondary structural elements of uncapped mRNA are of key interest in elucidating its enzymatic mechanism.Here, constructs of tobacco etch virus (TEV) mRNA, each containing specific structural elements, were used to study equilibrium binding. Steady state fluorescence spectroscopy was used to study binding between PAP and three distinct mRNA constructs. The Kd values for the TEV mRNA constructs are given here: Full length 5′‐leader (3 pseudoknots) (FL) = 63nM, PK1 (pseudoknot 1) = 183nM and S2‐3 (a mutation in PK1) = 85nM. These results suggest that PAP recognizes specific structural elements within the mRNA for binding. The data here will be compared to equilibrium binding between PAP and FL, PK1, and S2‐3 capped TEV mRNA. This will show whether the cap structure recruits PAP at a higher affinity than the uncapped mRNA in order to place PAP within the molecular reading frame for scanning and and possibly subsequent depurination.
Kinetic Analysis of Iron Responsive Element (IRE) mRNA with Iron Regulatory Protein (IRP1)
Authors: KHAN MATEEN A , Walden William E , Theil Elizabeth C , Goss Dixie J 
Published: June 2021

In mRNA a family of 5′‐noncoding structures, iron response elements (IRE), control mRNA translation. When iron concentrations decrease, IRP protein binds to the IRE‐RNA and inhibits formation of polysomes. To gain insight into the dynamic process of IRE‐mRNA/IRP regulation and the mechanism of binding, the kinetics of IRE mRNA binding to IRP1 protein were examined. The association rate constant for Ferritin IRE‐RNA was about 6‐fold faster as compared to mitochondrial IRE‐RNA (kon = 400 μM−1s−1 for Fer‐IRE and kon = 50 μM−1s−1 for mt‐IRE). However, the dissociation rate constant was similar for the two IRE‐RNAs binding to IRP1 protein. Addition of 50 μM magnesium ions (a model for air sensitive Fe2+) decreased the rate of association 5‐fold and increased the rate of dissociation 3‐fold for Ferritin IRE‐RNA and for mitochondrial IRE‐RNA the rate of association decreased <3‐fold and the rate of dissociation increased ~2‐fold. Metal ion selective destabilization of Ferritin and mitochondrial aconitase RNA/protein complexes as reported here explain in part quantitative differences in signal response to iron in vivo and indicate possible new regulatory interactions. These results demonstrate the first direct kinetic measurements of IRP1 protein binding to IRE‐RNA.NSF: MCB‐ 0814051 (to, D.J.G.)NIH: DK20251 (to E.C.T.)
Quantitative interactome analysis of RGS9‐2 degradation mechanisms in mammalian brain.
Authors: Posokhova Ekaterina , Uversky Vladimir N , Martemyanov Kirill A 
Published: June 2021

RGS9‐2 is a striatal‐specific protein involved into regulation of movement and reward through negative regulation of muopioid and D2 dopamine receptor signaling. Changing RGS9‐2 expression is an important way to achieve the plasticity of signaling regulation and behavioral outcomes. R7 binding protein (R7BP) is an interaction partner of RGS9‐2 that sets the expression of RGS9‐2 by controlling proteolytic stability of the complex. Uncoupling of RGS9‐2 from R7BP is thought to be a mechanism of RGS9‐2 levels regulation.The aim of the study was to elucidate the molecular mechanism of RGS9‐2 degradation upon loss of R7BP.We developed a proteomic approach for quantitative analysis of interactome changes in mouse knockout models and used it to identify interactions of RGS9‐2 upregulated in R7BP knockout mice. The screen resulted in the identification of 21 proteins among which chaperone Hsc70 was found to exhibit one of the biggest changes in binding to RGS9‐2. The interaction was validated both in vivo and in transfected 293 cells and was found to be mediated by the intrinsically disordered C‐terminal domain of RGS9‐2. Down‐regulation of Hsc70 expression using siRNA and a dominant negative approach increased levels of RGS9‐2.We conclude that Hsc70 is a negative regulator of RGS9‐2 levels in the striatum.NIH NIDA DA021743
DNA Analysis Using Polymerase Chain Reactions and Micro‐Electrophoresis
Authors: Hunley Ryan Timothy , Harwood John , Boles Jeffrey O. , Mancuso Matt 
Published: June 2021

Polymerase Chain Reaction (PCR) is used in a variety of applications in forensics, medicine, and research. PCR relies on the ability to thermally break apart a DNA strand, and replicate selected portions of the DNA (if present) through addition of “primer” coding for the selected DNA segment. Following amplification through 30–40 precisely controlled thermal cycles, samples are analyzed for the DNA segments by electrophoresis. We use a bioanalyzer “lab‐on‐a‐chip” instrument in the analysis, which represents an example of a micro‐total analysis system (u‐TAS).We are developing experiments to help chemistry students in the second semester quantitative analysis course become more familiar with PCR and with micro‐total analysis analyzers. We use a commercially available kit to provide needed PCR primers and reagents, and standard reference corn and soya powder samples containing the gene modifications. Components of “Round‐up Ready” and Bt‐producing crops, the 35S promoter of the cauliflower mosaic virus and the terminator of the nopaline synthase gene of Agrobacterium tumefaciens, are amplified. Students are assigned to bring in food samples of their own choosing for analysis. We are investigating the feasibility of performing analyses for other gene modifications in foods as experiments in undergraduate analytical and biochemistry courses.
Zebrafish as a model to determine the mechanisms of vitamin E function
Authors: Miller Galen W , Labut Edwin M , Lebold Katie , Floeter Abby , Tanguay Robert L , Traber Maret G 
Published: June 2021

Vitamin E (alpha‐tocopherol, AT) is required to prevent fetal resorption in rodents. To study its role in fetal development, a non‐placental model is required. Therefore, we have developed an AT‐deficient zebrafish model by feeding a defined diet (with or without AT) that we created. Zebrafish (age: 1 m) were fed the deficient (E−), sufficient (E+), or Lab diet up to 1 y. All groups showed similar growth and normal development. When compared to E+, the E− fish had lower (p<0.05) AT levels within 3 w; by 112 d the E− fish (3±1 nmol/g) contained 100 fold less AT than the E+ fish (375±100, p<0.001). The rate of depletion was 0.029 ± 0.006 pools/d. At age ~4 m, E‐depleted adults were spawned and produced viable embryos with similar AT concentrations to those of their parents. The E− embryos exhibited a higher mortality at 24 h post fertilization (hpf) and a higher combination of malformations and mortality at 120 hpf than the control diets. This study documents for the first time that vitamin E is essential for normal embryonic development. It also shows that the zebrafish is an exceptional model to define the molecular functions of vitamin E.Grant Funding Source: NIH #HD062109 and #ES00210
Endothelial protective effects of erythritol
Authors: den Hartog Gertjan , Berger Alvin , Bast Aalt 
Published: June 2021

Vascular health is strongly dependent on endothelial state, which regulates vascular tone and inflammatory responses. High glucose‐induced endothelial cell apoptosis, likely via reactive oxygen species (ROS), has a pivotal role in diabetes‐associated vascular diseases, including atherosclerosis. We evaluated the anti‐oxidative and endothelium‐protective properties of erythritol (ERT), a non‐caloric sweetener which is well tolerated, and whose chemical structure resembles mannitol, a hydroxyl radical scavenger. In competition assays, ERT was an excellent hydroxyl radical scavenger (inert towards superoxide radicals); and inhibited radical–induced hemolysis. ERT reacted with hydroxyl radicals forming erythrose and erythrulose by carbon‐bound hydrogen atom abstraction. In streptozotocin‐induced diabetic rats, ERT had endothelium‐protective effects and erythrose was detected in urine. The clear endothelial protective effects of ERT cannot be explained by hydroxyl radical scavenging only. We are currently investigating mechanisms underlying protective effects of ERT on endothelium dysfunction in various model systems.This research project was financially supported by Cargill Inc.
Press Juices from Hamour and Kingfish exhibit Antioxidant Properties
Authors: Waly Mostafa I , Al‐Sabahi Jamal N , Al‐Bulishi Mohamed , Soussi Bassam 
Published: June 2021

Many recent studies have suggested that regular consumption of antioxidant‐rich foods improve the cellular antioxidant capacity and hence reduce oxidative stress and related chronic diseases occurrence.The main aim of this study was to evaluate the antioxidants contents of the press juices extracted from Hamour and King Fish, locally available fishes in Oman.Press juices (PJs) filtrate was prepared by centrifuging the fish's light muscle portion at 18368 g for 2 hrs at 4 °C. For the measurements, 10 filtrates were prepared. Using HPLC, vitamin C was measured for Hamour PJ (89.22 μM ± 1.2) and for kingfish PJ (85.13 μM ± 1.2), p >0.05.Total antioxidants content was quantified using a commercial kit (RANDOX, UK) and the measurements were significantly higher for Hamour PJ (1.32 mmol/Liter ± 0.6) as compared to kingfish PJ (0.55 mmol/liter ± 0.8), p <0.05. Glutathione (GSH) was measured using fluorescent ELISA (APO‐GSH, USA) assay kit and the GSH content of Hamour fish PJ; 412.5 μg ± 9.3 was significantly higher than that of kingfish 269.43 μg ± 13.7, p <0.05. All results were the mean ± SD and student t‐test, unpaired test, was used for the statistical analysis.This study highlights the potential of aqueous of PJs filtrates from Hamour and kingfish as natural powerful antioxidants. Human cell culture studies are on‐going to shed further light on the protective health aspects of Press Juice.This study was supported by HM Trust Fund for Strategic Research, SR/AGR/FOOD/05/01.
Zip5: a candidate protein for zinc transport from maternal circulation into the mammary gland
Authors: Velasquez Vanessa Renee , Kelleher Shannon L 
Published: June 2021

An extraordinary amount of zinc (Zn) moves through the mammary gland (MG) and into milk during lactation, but the mechanism by which the MG accumulates Zn from maternal circulation is not yet understood. We postulate that a Zip protein is responsible for importing Zn into the MG. Zn accumulation in the MG in vivo was visualized using X‐ray fluorescence microscopy. Candidate Zip proteins (Zip1, 3, 5, 6, 8, 10) were identified by real‐time PCR in secreting mouse mammary epithelial cells (HC11). This screen identified Zip5 by a 10‐fold increase in mRNA expression relative to non‐secreting cells. To confirm Zip5 protein expression during lactation, we assessed expression by immunblotting. Zip5 protein expression was ~3 fold higher in lactating compared with non‐lactating MG, further supporting the hypothesis that Zip5 is important for Zn accumulation in the MG during lactation. To confirm that Zip5 was appropriately localized to transport Zn from maternal circulation, we successfully localized Zip5 to the serosal membrane of epithelial cells in mouse MG using immunohistochemistry, and by cell surface biotinylation and confocal microscopy in HC11 cells. In conclusion, our data supports the postulate that Zip5 is a candidate for Zn accumulation from maternal circulation as it is localized appropriately and protein expression increases during lactation in the MG.Supported by intramural funds to SLKGrant Funding Source: intramural funds to SLK
The effect of soy diets on obesity‐related pathologies in lean and obese Zucker rats.
Authors: Cain James , Davis Jeremy , Strader April , Butteiger Dustie 
Published: June 2021

The purpose of this study was to compare the effects of soy versus dairy whey and caseinate diets on obesity‐related pathologies such as non‐alcoholic fatty liver and impaired glucose homeostasis in an insulin resistant rodent model. Male lean (L) and obese (O) Zucker rats were randomly assigned to three diet groups for 17 weeks: sodium caseinate control (C), dairy whey control (W) and soy protein (S). Physiological data were collected throughout the study and at week 14 animals were subjected to an oral glucose tolerance test (OGTT). As previously shown, OS animals had greater final body weights and body fat percentages, and exhibited an increased food intake vs the caseinate group. OS rats had a significantly lower liver weight and percent liver lipid content versus OC and OW groups. There was no significant difference in non‐fasted blood glucose levels or OGTT between diet groups. Nonetheless, OS rats had significantly lower OGTT plasma insulin concentrations at 15 and 30 min compared with OC. OS and OW groups had significantly lower insulin:glucose ratios than OC at all OGTT time points and for AUC, indicating higher insulin sensitivity in these groups. No statistically significant differences were identified in the lean groups. Further molecular and biochemical analyses are currently under examination. Preliminary results demonstrate benefits of a soy protein diet in amelioration of obesity‐related pathologies such as non‐alcoholic fatty liver and impaired insulin sensitivity.Grant Funding Source: ISA
Comparison for bioavailability of 1‐deoxynojirimycin with mulberry water extract in rat
Authors: Kim Ji Yeon , Kwon Hye Jin , Jung Ji Young , Kim Do Ram , Kim Young Suk , Kwon Oran 
Published: June 2021

The bioavailability of dietary phytochemicals may be influenced by the food matrix in which they are prepared. In this study we compared the intestinal absorption and excretion of orally administered 1‐deoxynojirimycin, which is well known as a potent glucosidase inhibitor, as isolated DNJ and mulberry water extract. When the rats sacrificed after administered mulberry water extract or isolated DNJ at 0, 15, 30, 45, 60, 120, 240, and 360 min, plasma DNJ levels reached a maximum at 30 min and then rapidly decreased. The DNJ levels of plasma 30 min after administration of mulberry leaf extract were lower than isolated DNJ with statistical significance (p<0.05). Moreover, the DNJ was dose‐dependently absorbed from the digestive tract into rat plasma (p<0.0001). We analyzed the concentration of DNJ in plasma, feces, and urine when administered as isolated DNJ as well as mulberry water extract. The absorption and excretion pattern was similar between them. These findings indicate that orally administered mulberry water extract and isolated DNJ are absorbed and excreted as similar pattern and the absorption rate for isolated DNJ is higher than water extract.Grant Funding Source: ASN
Type 2 Diabetes‐associated changes in the plasma lipidome in obese women
Authors: Grapov Dmitry , Adams Sean H , Garvey W. Timothy , Lok Kerry H , Newman John W 
Published: June 2021

Type 2 diabetes (T2D) is associated with elevated plasma fasting glucose, glycosylated hemoglobin (HbA1c), non‐esterified fatty acids (NEFA) and endocannabinoids (EC). An expanded assessment of lipidomic changes in diabetes is hypothesized to provide novel insights into metabolic changes associated with this malady. Targeted profiling of plasma lipids was performed in 43 diabetic and 12 non‐diabetic obese women. Plasma ECs (n=35), oxylipids (OxL; n=80), and NEFA (n=54) concentrations were measured. Multivariate statistics and regression analyses were used to mine the data matrix. Multivariate models showed a diverse array of NEFA, ECs, EC‐like compounds and OxLs to be significantly altered in the diabetic state. As opposed to the independent variables, an algorithm using subject age, BMI, HbA1c and fasting glucose was normally distributed and also segregated diabetic and non‐diabetic subjects. A linear model using measured lipids was developed that predicts the discriminating metric (adjR2=0.82) driven by OxL and EC subcomponents. Finally, this model showed that diabetic subjects had shifts in plasma levels of P450‐mediated arachidonic acid metabolites known to influence tissue plasminogen activator expression, which may parallel known shifts in vascular homeostasis.This work was supported by USDA‐ARS Project 5306‐51530‐016‐00D, NIH‐NIDDK R01DK078328‐01 and T32‐GM08799.
Validated biomarkers of caloric restriction in rats: Markers of disease risk in humans?
Authors: Kristal Bruce , Matson Wayne , Willett Walter , Hankinson Susan , Shurubor Yevgeniya , Sheldon Diane , Krasnikov Boris , Russell Neil , Sniatynski Matt , Marur Vasant 
Published: June 2021

Caloric restriction (CR) robustly increases longevity/reduces morbidity in mammals, eg., breast cancer risk is generally decreased >90% in CR rodents. CR is usually dominant to other risk factors and are directly analogous to human data linking obesity with poor health outcomes, including cancer. We tested if biomarkers of diet in rats predicts future disease in humans.MethodsMetabolomics measurements in sera/plasma were conducted w/HPLC/Coularray detectors (N~600 rats, ~1700 humans). Classification/predictive power were tested, optimized, and validated using a series of megavariate data analysis approaches in sequential blinded cohorts.ResultsModels distinguish diet groups with >95% accuracy. Diets varying in extent/duration of CR were used to develop models for intermediate caloric intakes. Markers were adapted for human study, analytically validated at instrumentation (N=30; 100% blinded split accuracy) and sample collection levels (N=34; most stable under worst case conditions), then biologically validated (N~70 in triplicate, metabolites/profiles had intraclass correlation coefficients ~0.65–0.85). Marker profile scores reflects energy intake/risk of future breast cancer in 210 case/control pairs nested within the Nurses’ Health Study.ConclusionMetabolomics profiles offer a potential biochemical approach to validate nutritive status and contribute to risk analysis. NIH