bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Heat Shock Protein 70 is a Novel Target for Nobiletin in Human Colon Cancer Cells
Authors: Gao Zili , Zheng Jinkai , Wu Xian , Dimova Kalina , Scordilis Stylianos , Xiao Hang 
Published: June 2021

Nobiletin is a unique flavonoid mainly found in citrus fruits, and has been reported to inhibit colon carcinogenesis in multiple rodent models. However, the direct molecular targets of nobiletin are unknown. Heat shock protein 70 (HSP70) contributes to cancer cell survival and resistance to chemotherapies, thereby inhibition of HSP70 is a promising strategy in cancer chemoprevention. Using affinity chromatography, we demonstrated that nobiletin bound to HSP70 at its ATP‐binding domains in human colon cancer cells and specifically inhibited its ATPase activity. Down regulation of HSP70 by siRNAs significantly increased cancer cells’ sensitivity to nobiletin treatments, which were evidenced by increased apoptosis and decreased colony formation capacity of cancer cells. The association between HSP70 and HSP90 is critical for the stability of their client proteins, such as EGFR, AKT, and STAT3, which contributes to carcinogenesis. We found that nobiletin disrupted the association between HSP70 and HSP90, and combined treatment with nobiletin and HSP90 inhibitors promoted the degradation of those client proteins. Moreover, combined treatment with nobiletin and HSP90 inhibitors resulted in enhanced inhibition on cancer cell growth by inducing apoptosis and cell cycle arrest. Most interestingly, our results also demonstrated that the major colonic metabolites of nobiletin (generated in the colon of mice fed with nobiletin) showed similar inhibitory effects against HSP70‐mediated signaling pathways. Overall, our results demonstrated, for the first time, that HSP70 is a promising target of nobiletin and its colonic metabolites in inhibiting human colon cancer cell growth, which warrants further investigation on their role in colon cancer chemoprevention.Support or Funding InformationThis study was supported by funding from NIH, AICR and USDA.
Dietary quercetin inhibits methylglyoxal‐induced advanced glycation end products formation in mice
Authors: Zhao Yantao , Wang Pei , Chen Huadong , Sang Shengmin 
Published: June 2021

Increasing evidence has identified the formation of advanced glycation end products (AGEs) as a major pathogenic link between hyperglycemia and diabetes‐related complications. As the precursor of AGEs, methylglyoxal (MGO) is observed at relative higher plasma levels (2–6 fold) in diabetic patients than in healthy subjects and are also found in many food products, beverages and cigarette smoke. Chronic intake of exogenous MGO has been demonstrated as one of the cause factors for the development of diabetes. Therefore, decreasing the levels of MGO will be an effective approach to reduce the formation of AGEs and the development of diabetic complications. In this study, we investigated whether long‐term treatment of MGO in drinking water can cause protein glycation in mice and whether dietary flavonoids, such as quercetin, can prevent the formation of AGEs via trapping MGO. Our results indicate that chronic MGO administration (0.06% in drinking water for 20 weeks) significantly elevates the levels of MGO and AGEs in mouse plasma, liver and kidney, and cause liver injury. Dietary quercetin treatment (0.067% and 0.2% in diet) ameliorates MGO‐induced liver injury and prevents the formation of AGEs at least in Kidney. Further mechanistic studies demonstrate that both quercein and its major metabolite isorhamnetin could trap MGO in mice to form mono‐ and di‐MGO adducts. In addition, quercetin could increase glyoxalase I and II expression and GSH/GSSG ratio to promote the detoxification of MGO.Support or Funding InformationUSDA‐AFRI grant 2012‐67017‐30175 to S. Sang
Using Synthetic Biology methods to construct a rapid and efficient estrogen biosensor
Authors: Lynott Michaela , Huttelmaier Stefanie , Tobiason Deborah , Canfield Jennifer 
Published: June 2021

Synthetic Biology demonstrates the integral relationship between biology, chemistry, mathematics and computer science. The field provides innovative approaches to a wide range of applications, such as bioremediation, sustainable energy production and biomedical therapies. Synthetic biology involves construction of genetic circuits when DNA sequences are modified to include standard restriction enzyme sites making the DNA sequences interchangeable and allowing for construction of a variety of circuits. New tools and devices are constantly needed to enhance the already extensive list of BioBricks housed at the Registry of Standard Biological Parts. As proof of concept we constructed a new BioBricks based on the toluene sensor built by Stiner and Haverson (AEM, 2002) in Pseudomonas. Toluene is an environmental contaminant that is the byproduct of oil contamination. Biosensors generated from BioBrick parts yield more affordable, transportable, and faster methods of pollutant detection. The tbuT cassette from Stiner and Haverson's work was digested with XbaI and SpeI sites then ligated with the GFP reporter gene and cloned into iGEM BioBrick vectors in order to create a biosensor in Escherichia coli. These BioBricks can be readily used with a variety of reporter genes to create new versions of the biosensor. We expanded our biosensor construction to create a dual functional estrogen sensitive BioBrick. The presence of estrogenic compounds (endocrine‐disruptors, EDCs) in the water supply raises concerns about human and aquatic health. Current methods for detecting estrogen contamination require expensive, time‐consuming techniques such as liquid chromatography‐mass spectrometry and high performance liquid chromatography. Previously reported estrogen biosensors required multiple cloning and transformation steps for successful detection in bacteria. Our estrogen responsive plasmid contains parts that allow for both the constitutive expression of the estrogen receptor followed by an estrogen response element toggled to a reporter gene in a single vector. When the estrogen response element is activated the reporter gene, red fluorescent protein, will produce a color change in E.coli. We will present data demonstrating the efficiency of this dual‐functional biosensor and it effectiveness for estrogenic compounds in contaminated water.Support or Funding InformationNSF grant #NSF‐GPG‐1444406, GPG: Research Collaboration: REU Pilot: Synthetic Biology Boot camp
Structural and Biochemical Characterization of AidC, a Quorum‐Quenching Lactonase With Atypical Selectivity
Authors: Mascarenhas Romila Nina 
Published: June 2021

Quorum‐quenching catalysts are of interest for potential application as biochemical tools for interrogating interbacterial communication pathways, as antibiofouling agents, and as anti‐infective agents in plants and animals. Herein, the structure and function of AidC, an N‐acyl‐l‐homoserine lactone (AHL) lactonase from Chryseobacterium, is characterized. Steady‐state kinetics show that zinc‐supplemented AidC is the most efficient wild‐type quorum‐quenching enzymes characterized to date, with a kcat/KM value of approximately 2 × 106 M−1 s−1 for N‐heptanoyl‐l‐homoserine lactone. The enzyme has stricter substrate selectivity and significantly lower KM values (ca. 50 μM for preferred substrates) compared to those of typical AHL lactonases (ca. >1 mM). X‐ray crystal structures of AidC alone and with the product N‐hexanoyl‐l‐homoserine were determined at resolutions of 1.09 and 1.67 Å, respectively. Each structure displays as a dimer, and dimeric oligiomerization was also observed in solution by size‐exclusion chromatography coupled with multiangle light scattering. The structures reveal two atypical features as compared to previously characterized AHL lactonases: a “kinked” α‐helix that forms part of a closed binding pocket that provides affinity and enforces selectivity for AHL substrates and an active‐site His substitution that is usually found in a homologous family of phosphodiesterases. Mutations on this His substitution are being studied along with other suspected catalytic residues. Implications of the catalytic mechanism of AHL lactonases will be discussed based on these data and prior knowledge of other AHL lactonases.Support or Funding InformationNational Science Foundation, Grant No. CHE‐1308672
Dynamic Stiffness Reveals New Cardiac Phenotypes in Patient‐Derived iPSCs
Authors: Engler Adam J. 
Published: June 2021

After nearly a decade of recognition that extracellular matrix (ECM) properties can influence cell behavior to similar degree as growth factors, in particular ECM composition, topography, porosity, and stiffness, biologists have come to recognize its importance. However matrix is highly dynamic, changing how much of it is secreted and assembled during development and disease. Most synthetic ECM mimics made initially to study this phenomenon were static but there is growing interest in making matrices that have tunable properties with time. To accomplish this, my lab has created hyaluronic acid (HA)‐based materials that crosslink themselves spontaneously or via UV‐sensitive reactions to provide slow or “on‐demand” changes in stiffness. By knowing the dynamics of matrix assembly during heart development as well as heart remodeling post‐heart attack, we have used these HA materials to model how the heart tube stiffens or the onset of fibrosis (i.e. “heart attack‐in‐a‐dish”), respectively, to understand how temporal changes in stiffness better instruct differentiation and function. Using an embryonic heart model, we have found that developmentally appropriate stiffening can drive maturation and sarcomere assembly in pre‐cardiac mesoderm. Conversely using patient‐derived induced pluripotent stem cells (iPSCs) in HA that models a “heart attack‐in‐a‐dish,” we have identified new cardiac phenotypes in diseased cardiomyocytes that were previous masked by other conditions in patients. We have also used the materials to study cardiac disease mechanisms, since often phenotypes do not appear until cells are stressed by the “heart attack‐in‐a‐dish” conditions. Based on these exciting results, we believe that any in vitro culture system should employ dynamic materials that change as the niche does in vivo.Support or Funding InformationNIA/NIH, NHLBI/NIH, NSF, and AHA
Using Skin Carotenoids to Assess Potential Dietary Changes After One Academic Year in the Shaping Healthy Choices Program
Authors: Nguyen Lori M , Scherr Rachel E , Dharmar Madan , Ermakov Igor V , Gellermann Werner , Jahns Lisa , Linnell Jessica D , Keen Carl L , Miyamoto Sheridan , Steinberg Francene M , Young Heather M , Zidenberg‐Cherr Sheri 
Published: June 2021

Reported dietary intake is often used in community interventions to assess intake of fruits and vegetables (F/V); however, dietary assessment methods are inaccurate, and time and labor intensive. Skin carotenoids are a potential biomarker to assess F/V intake given that carotenoids are predominately found in F/V and their concentration in the skin can reflect dietary intakes of F/V. The Resonance Raman Spectroscopy (RRS) method is a quick and validated method to assess skin carotenoids. The objective of this study was to determine whether skin carotenoids measured by RRS can be used to confirm changes in dietary intake of F/V in 4th‐grade students participating in the Shaping Healthy Choices Program (SHCP). The SHCP is a multi‐component nutrition intervention and one of its aims is to increase F/V intake. Dietary intake was assessed using the Block Food Frequency Questionnaire in a subset of students (n=30) and skin carotenoids were measured using RRS (n=82) before and after the SHCP intervention. Pre‐post changes in F/V intake measured by each method were tested using Student's t‐tests. After 9 months, reported intake of carotenoids decreased by 19% (1469mcg, P=0.05) and skin carotenoids decreased by 9% (2882 RRS intensity units, P<0.001). Change in reported intake correlated with change in skin carotenoids (r=0.43, P=0.02). While the reported decrease in F/V intake in this population was unanticipated given that the intervention aimed to increase intake, the RRS measurements confirmed this change. In conclusion, optically measured skin carotenoids can be used to help evaluate changes after 9 months of F/V intake in 4th‐grade students.Support or Funding InformationFunding was provided by UCANR competitive grant #11‐1018, USDA 2011‐38420‐20082, and USDA NIFA, Hatch project 221082.
Vitamin B12 and Folate Status in Pregnant Adolescents and Their Infants
Authors: Finkelstein Julia L. , Guillet Ronnie , Pressman Eva K. , Kent Tera R. , O'Brien Kimberly O. 
Published: June 2021

ObjectiveTo determine the prevalence of vitamin B12 deficiency and examine the associations between maternal and infant vitamin B12 and folate status in a cohort of healthy pregnant adolescents.DesignSerum vitamin B12 and folate concentrations were measured in pregnant adolescents at mid‐gestation, delivery, and in cord blood. Linear and binomial regression models were used to examine the associations between maternal and infant vitamin B12 and folate status.ResultsSerum vitamin B12 concentrations decreased significantly during pregnancy (p<0.0001) resulting in an increased prevalence of vitamin B12 deficiency (<148 pmol/L) from mid‐gestation (1.6%) to delivery (15.3%). At delivery, an additional 46.6% of mothers were vitamin B12 insufficient (<200 pmol/L). Maternal vitamin B12 concentrations (p<0.001) and vitamin B12 deficiency (p<0.001) at delivery were associated with infant B12 concentrations, with 1% of infants being vitamin B12 deficient at birth. Similarly, maternal folate concentrations at delivery (p<0.001) were associated with infant folate concentrations.ConclusionsMaternal vitamin B12 and folate status at delivery predicted infant vitamin B12 and folate status in healthy pregnant adolescents.Support or Funding InformationSupported by grants from the United States Department of Agriculture (USDA): USDA 2005‐35200‐15218 and USDA 2010‐34324‐20769; and the Division of Nutritional Sciences, Cornell University
A High Flavonoid Diet Does Not Improve Markers of Insulin Resistance and Subclinical Inflammation in Overweight and Obese Men and Women
Authors: Lefevre Michael , Ward Robert , Bergeson Janet , Hergert Nancie , Hergert John , Aguilar Sheryl 
Published: June 2021

Populations consuming healthy diets high in flavonoids have lower levels of subclinical inflammation, better insulin sensitivity and reduced risk for both cardiovascular disease and type 2 diabetes. We hypothesized that providing a diet high in flavonoids would reduce insulin resistance and subclinical inflammation relative to a low flavonoid diet. To test this hypothesis, 29 overweight/obese (BMI; 25.2–34.5 kg/m2) men (N=16) and women (N=13) were provided two well‐controlled diets that were identical in macronutrient content (Protein, 17% en; Fat, 30% en; Carbohydrate, 53% en), but differed markedly in flavonoid content (Low Flavonoid Diet, 10 mg/1000 Kcals; High Flavonoid Diet, 340 mg/1000 Kcals). All meals for both diets were prepared and fed for 6 weeks each in a randomized cross‐over design with endpoints determined in duplicate during the last week of each diet period. All meals were well tolerated by the participants and overall compliance as assessed by questionnaires and direct observations was excellent. Relative to the Low Flavonoid Diet (LFD), the High Flavonoid Diet (HFD) did not significantly affect serum chemistry (blood urea nitrogen, creatinine, estimated glomerular filtration rate, sodium, potassium, chloride, CO2, calcium, total protein, albumin, globulin, A/G ratio, total bilirubin, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase). Surprisingly, the HFD had no significant effects on blood pressure and plasma lipids (triglycerides, LDL‐C, HDL‐C) and tended to increase (all non‐significant) markers of inflammation (C‐reactive protein, soluble TNFα receptor‐1, soluble TNFα receptor‐2, interleukin‐6). Contrary to expectations, markers of insulin resistance (fasting insulin, glucose, HOMA‐IR) were modestly, but significantly higher on the High Flavonoid Diet relative to the Low Flavonoid Diet. Under the conditions of this study, diets high in flavonoids failed to improve plasma lipids, insulin sensitivity or subclinical inflammation. Our results are at odds with the reported beneficial effects of high flavonoid diets on CVD risk factors, insulin sensitivity and systemic inflammation observed in both preclinical and epidemiological studies.Support or Funding InformationThis work was supported by USDA/NIFA grant 2014‐67017‐21749.
Dietary Glycomacropeptide Increases Fat‐Free Mass, Bone Mineral Content and Fatty Acid Oxidation Compared With Casein In Female Mice
Authors: Sawin Emily Ann , Stroup Bridget Marie , Murali Sangita G , Ney Denise Marie 
Published: June 2021

BackgroundGlycomacropeptide (GMP) is a 64 amino acid (AA) glycophosphopeptide released from κ‐casein during cheese making that promotes satiety, reduces body fat, increases bone mass and infers prebiotic and anti‐inflammatory effects.ObjectiveTo determine if GMP, when fed as the primary protein source, modulates feed efficiency, weight gain and body composition and increases fat oxidation compared with casein.MethodsWeanling female C57Bl6 mice were fed high and low fat diets (HF 60% en; LF 13% en) with casein or GMP for 28 wks with assessment of body weight and food intake. Body composition was assessed by DXA. Fatty acid oxidation was measured in liver, muscle, and fat tissues using 14C‐palmitate. Plasma concentrations of amino acids, hormones and cytokines were determined.ResultsDespite similar food intake, mice fed the HF GMP diet had significantly lower final body weight than mice fed the HF casein diet. Fat free mass was increased in mice fed the LF GMP diet relative to the LF casein diet with similar final body weight but greater food intake. Regardless of dietary fat content, GMP significantly increased bone mineral content (BMC) and fatty acid oxidation in subcutaneous fat compared to casein.ConclusionsGMP reduces feed efficiency, modulates body composition to increase fat free mass and BMC, and increases fat oxidation compared to casein in female mice. GMP is a bioactive food ingredient with application to obesity and osteopenia in females.Support or Funding InformationSupported by the Wisconsin Alumni Research Foundation, NIH T32 DK 007665, and USDA Hatch WIS01790.
Relationship between concentrations of lutein and StARD3 among pediatric and geriatric human brain tissue
Authors: Johnson Elizabeth J , Tanprasertsuk Jirayu , Li Binxing , Bernstein Paul S , Vishwanathan Rohini , Johnson Mary Ann , Poon Leonard W 
Published: June 2021

Lutein, a dietary carotenoid, selectively accumulates in human retina and brain. While many epidemiological studies show evidence of a relationship between lutein status and cognitive health, lutein's selective uptake in human brain tissue and its potential function in early neural development and cognitive health have been poorly evaluated at a molecular level. The objective of this study was to evaluate the cross‐sectional relationship between concentrations of brain lutein and StARD3 (identified as its binding protein in retinal tissue) among three age groups: infants (1–4 months, n=10), older adults (55–86 years, n=8), and centenarians (98–105 years, n=10). Brain lutein concentrations were analyzed by high‐performance liquid chromatography and StARD3 levels were analyzed by Western Blot analysis. The strong relationship in infant brains (r=0.75, P <0.001) suggests that lutein has a role in neural development. The relationship remained significant but weaker in older adults (r=0.51, P <0.05) and insignificant in centenarians (r=0.08, P >0.05), eight of whom had dementia. Although the sample size for each group was small, these results suggest an age‐related decrease or abnormality of StARD3 activity in human brain. Given that StARD3 is also involved in cholesterol transportation, a process that is aberrant in neurodegenerative diseases, the potential protective function of lutein against these diseases remains to be explored.Support or Funding InformationThis work was supported by : USDA agreement 58‐1950‐4‐003, NIH 1PO1‐AG17553, Abbott Nutrition, DSM Nutritionals, NIH EY‐11600 (PSB), NIH EY‐14800 (PSB) and an unrestricted grant to the Moran Eye Center of the University of Utah from Research to Prevent Blindness
Differential Expression of Genes Involved in Inflammatory Immune Response and Protein Ubiquitination in the Prefrontal Cortex of Rhesus Macaque with High and Low Lutein Content
Authors: Mohn Emily S , Erdman John W. , Neuringer Martha , Kuchan Matthew J , Johnson Elizabeth J 
Published: June 2021

Lutein, a carotenoid with antioxidant and anti‐inflammatory functions, accumulates in the primate brain. Studies have shown that brain levels of lutein are more consistently related to better cognitive test scores in humans than other antioxidants found in higher concentrations in the brain, such as vitamin E. Supplementation with lutein can improve verbal fluency, memory scores, and rates of learning in older adults. However, the mechanism by which lutein functions in cognition is unclear. The aim of this study was to determine the relationship between lutein concentration and gene expression profiles in rhesus macaque brain. Brain tissue was dissected from monkeys fed a chow diet containing lutein (~16 μmol/kg diet). Next generation RNA sequencing was used to determine differentially expressed genes in the prefrontal cortex of monkeys (11–13y) with high lutein content (mean=34.5 pg/mg wet weight, n=3) versus those with low lutein content (mean=11.8 pg/mg wet weight, n=3). Lutein concentrations were determined in these brain samples using HPLC. A total of 415 genes were differentially expressed between the two groups. Ingenuity Pathway Analysis (IPA) revealed the up‐regulation of pro‐inflammatory immune response pathways, such as chemokine signaling, antigen presentation, leukocyte extravasation, and neuropathic pain signaling, in brains with low lutein content. A total of 16 genes were upregulated in this pro‐inflammatory network, with specific genes of interest including beta‐2‐microglobulin and p38 MAPK (1.5 fold change for each), which are both associated with age‐related cognitive impairment and neurodegeneration. Additionally, up‐regulation of the protein ubiquitination pathway (9 genes), a hallmark of Alzheimer's disease, was observed in brains with low lutein content. These results indicate that lutein may play a role in suppressing neuroinflammation at the transcription level, and provide the first steps towards understanding the mechanisms underlying the beneficial effect of lutein on cognition.Support or Funding InformationThis work was supported by grants from Abbott Nutrition through the Center for Nutrition, Learning, and Memory at the University of Illinois, USDA (58‐1950‐4‐003), and NIH grant P51OD011092 to the Oregon National Primate Research Center