bluuebunny/crossref_metadata_2025 huggingface dataset:
Formation of ω‐NG–monomethylarginine as the sole product of human protein arginine methyltransferase 7 (PRMT7): a true type III methyltransferase?
Authors: Lopez Cecilia I , Sandberg Troy , Clarke Steven G.
Published: June 2021

Methylaton of arginine residues is an important posttranslational modification involved in transcriptional regulation, mRNA splicing, and signal transduction. Mammalian protein arginine methyltransferases (PRMTs) have been found to generate three different products. Type I PRMTs form both ω‐NG–monomethylarginine (ω‐MMA) and asymmetric ω‐NG,NG ‐dimethylarginine (ADMA), type II enzymes form ω‐MMA and symmetric ω‐NG,N'G–dimethylarginine (SDMA), and type III enzymes form solely ω‐MMA. Although the first six PRMTs discovered were clearly either type I or type II enzymes, the seventh has proven difficult to characterize. Initially, full‐length human PRMT7 expressed in E. coli was found to generate only ω‐MMA residues in small peptides, suggesting this is a type III enzyme. Other researchers, however, later reported that PRMT7 is also capable of forming SDMA in both peptides and protein substrates, characterizing it as a type II enzyme. We have now analyzed a more active PRMT7 preparation with both peptides and proteins and see only type III activity, forming ω‐MMA. It is possible that the substrate specificity of PRMT7 may depend upon additional co‐factors or binding to additional polypeptides. This work was supported by an NRSA award to CZL (GM078761) and an NIGMS grant to SGC (GM026020).
Characterization of bovine 20a‐HSD gene promoter
Authors: Min Kwan‐Sik , Purevjargal Naidansuren , Kang Myeong‐Hwa
Published: June 2021

The 20¥á‐hydroxysteroid dehydrogenase (20¥á‐HSD) enzyme catalyzes the NADPH‐dependent reduction of progesterone to 20¥á‐hydroxypregn‐4‐ene‐3‐ one (20¥á‐hydroxyprogesterone), which is biologically inactive. Thus, it is very important enzyme to maintenance of pregnancy in mammlian. We cloned the promoter (−927bp) of bovine 20a‐HSD gene to charaterize the functional role and constructed five deleted mutants into the pGL3‐basic vector. And we also constructed the EGFP gene vector under bovine 20a‐HSD promoter. Bovine 20a‐HSD gene promoter contains TATA box, two putative CCAAT boxes and several transcriptional factor sites (GATA‐1, CRE‐BP, Oct‐1, Ap‐1 and SRY). The promoter activity was remarkably decreased in all deleted vector using the rat corpus luteum cells.

But, it was recovery by the co‐transfection of the Sp‐1 factor. The constructed vector (promoter+EGFP) was expressed EGFP signal into the CHO K1 cell at 2 days after transfection.

(This work was supported by a grant (20080401034074) from BioGreen 21 Program, Rural Development Administration, Republic of Korea)
Allosteric Modulators of a Protein Kinase via Disulfide‐Trapping
Authors: Sadowsky Jack David , Wells James
Published: June 2021

Given the central roles kinases play in disease, much effort in academia and industry has focused on identifying small molecule kinase inhibitors. Relative to the ATP‐competitive inhibitors that usually result from these efforts, allosteric ligands often have higher selectivity and may regulate kinase activity in novel ways (e.g., via kinase activation or blocking protein‐protein interactions). We are using a site‐directed ligand discovery tool, disulfide‐trapping, to identify compounds that modulate kinase activity by binding to a specific allosteric site, the hydrophobic motif (HM) pocket. This site is functionally important in many kinases, but is not well conserved, suggesting that a large proportion of the kinome will be susceptible to modulation by HM pocket ligands and that such ligands will be selective for their targets. In exploring this concept, we have discovered both allosteric activators and inhibitors of PDK1, a kinase involved in cancer and diabetes. We have characterized the mechanism of action of these compounds using X‐ray crystallography and protein stability measurements; efforts to assess selectivity and apply these modulators in cells are ongoing.
Shape‐shifting proteins with a potential link to drug side effects
Authors: Lawrence Sarah H , Selwood Trevor , Ramirez Ursula D. , Stith Linda , Jaffe Eileen K
Published: June 2021

Human porphobilinogen synthase (huPBGS, EC 4.2.1.24) exists as an equilibrium of high activity octamers, low activity hexamers, and alternate dimer configurations that dictate the stoichiometry and architecture of further assembly. Reduced huPBGS activity factors in the disease states lead poisoning and ALAD porphyria, the latter of which involves hexamer‐favoring huPBGS variants. The Jaffe lab has established that small molecules can allosterically inhibit huPBGS activity by stabilizing the hexamer and putatively potentiate disease states. Two such inhibitors were identified by in silico docking to a hexamer‐specific surface cavity of huPBGS. The compounds were confirmed to inhibit wild‐type huPBGS activity by stabilizing the hexamer in vitro, and showed increased potency against two porphyria‐associated variants. One of the compounds is a structural analog to two amebicidal drugs, which have porphyria‐like side effects, and were found to stabilize the huPBGS hexamer in vitro. If the drugs can inhibit huPBGS by inducing hexamer formation in vivo, this would constitute an unprecedented explanation for a drug side effect. It is possible that other known drugs, toxins, or environmental contaminants could have a hexamer‐stabilizing inhibitory effect on huPBGS. An ongoing screen of ~1500 compounds approved for pharmaceutical use has so far revealed 6 compounds that stabilize the huPBGS hexamer in vitro.
Imaging the Distribution of Carbon‐11 Labeled Rifampicin, Isoniazid and Pyrazinamide in Baboons using PET
Authors: LIU LI , Hooker Jacob M. , Fowler Joanna S. , Tonge Peter J.
Published: June 2021

PET imaging studies of carbon‐11 labeled rifampicin (RIF), isoniazid (INH) and pyrazinamide (PZA) in baboons have been performed to provide more direct insight into pharmacokinetics and biodistribution of drugs for tuberculosis (TB) treatment and these baboon studies is a prelude to human imaging studies. These investigations would help us to determine better regime to treatment TB especially CNS TB because of its limited accessibility. It is also hypothesized that such high specific activity radiotracers may also useful for determining the location of bacterial population in vivo since these drug are expected to accumulated with in the bacteria. We find that all drugs cleared rapidly from lungs and injected [11C]RIF accumulates in gall bladder while injected [11C]INH and [11C]PZA accumulate in bladder. This is consistent with mechanism of excretion. We also find organ distributions of each drug are different from plasma PK, which is from several fold to hundreds fold. All of three injected drugs demonstrated higher concentration in lung than plasma over the study frame. Estimation based on the weight of baboon and normal dose and assumption of no metabolite suggested: concentration of RIF in lung is at least 5 times higher than MIC90, INH is more than 10 times higher than MIC90 and PZA is 1–3 times higher than MIC90 against MTB. The ability to enter the brain decreases in the order PZA > INH > RIF. Estimation of brain concentration based on same assumption as lung suggested that concentration of RIF is 3–4 times over MIC90 and INH is more than 10 times over MIC90, while PZA is quite close to MIC90 against MTB. These data have important implications for treatment of TB and CNS TB and set the scene for additional studies in humans.
Novel transformations in the biosynthesis of the marine‐derived antibiotic marinopyrrole
Authors: Ryan Katherine S , Gulder Tobias A.M. , Moore Bradley S
Published: June 2021

The marine‐derived natural product marinopyrrole has strong promise as an antibiotic, possessing activity against methicillin‐resistant Staphylococcus aureus (MRSA). Further, the chemical structure of marinopyrrole is unprecedented among natural products, consisting of two richly halogenated pyrrolic “monomers” dimerized to give an axially chiral bispyrrole structure [Org. Lett. (2008) 10: 629–631]. We are investigating the biosynthetic origins of this molecule with two purposes in mind. First, an elucidation of the biosynthesis of marinopyrrole will enable us to embark on bioengineering experiments to generate marinopyrrole derivatives with improved antibiotic properties. Second, an understanding of how the key chemical features of the molecule, including the axis of chirality and the halogenation pattern, are enzymatically generated will provide fundamental insights into the enzymology of this particular pathway and insights into antibiotic biosynthesis more generally. To isolate the biosynthetic gene cluster for marinopyrrole, we generated and screened a cosmid library of genomic DNA from the producing organism, and sequenced two cosmids containing overlapping portions of the marinopyrrole biosynthetic cluster. The assembled cluster of ~40 kbp consists largely of enzymes with high sequence identity with those found in the pyrrolomycin biosynthetic gene cluster [Antimicrob. Agents Chemother. (2007) 51: 946–957], in addition to candidates for catalyzing the unique coupling reaction.
Biophysical basis for TetX mediated antibiotic resistance
Authors: Walkiewicz Katarzyna , Lau Kelsey , Shamoo Yousif
Published: June 2021

We are studying the biophysical origins of drug resistance in order to define how changes at the molecular level lay the foundation for the overall success of an organism undergoing adaptation. Recently, a novel mechanism of tetracycline resistance was identified that inactivates tetracycline antibiotics via the bacterial enzyme TetX. To understand the biophysical basis for TetX mediated resistance, we propose to study the three dimensional structure of TetX using X‐ray crystallography. Full length crystals of TetX were obtained in space group P21 that diffracted to 2.8 Å resolution with unit cell dimensions 87.5 × 67.2 × 152.0 Å and β= 101.7°. Crystals of Seleno‐methionine containing TetX for use in Multi‐wavelength Anomalous Diffraction (MAD) are under optimization. In addition to biophysical studies, we have used experimental evolution to study how TetX facilitates resistance within a bacterial population. E. coli population containing TetX was adapted to high minocycline levels over the course of two weeks showing a >150 fold increase in the minimum inhibitory concentration (MIC). Our experiments indicate that TetX along with changes at the genomic level confer an increase in resistance that is highly synergistic and suggests a novel mechanism for resistance that extends beyond simple inactivation of tetracyclines. This work was supported by Robert A. Welch Foundation (C1584).
Associations among high prepregnancy body mass index, use of a breast pump and breastfeeding outcomes: Data from the Infant Feeding Practices Study II
Authors: Leonard Stephanie A , Labiner‐Wolfe Judy , Rasmussen Kathleen M
Published: June 2021

Although high prepregnancy body mass index (PPBMI, > 25 kg/m2) is associated with less successful breastfeeding (BF), it is unknown whether this extends to the use of a breast pump (BP). The associations among high PPBMI, use of a BP and BF outcomes were studied among the 2279 women in the Infant Feeding Practices Study II who ever breastfed; 1874 of these women used a BP. Data were analyzed by regression analysis in models adjusted for confounding factors. There was no difference between those with high or a normal PPBMI in whether a mother used a BP, was successful at expressing her milk if she did use a BP, why she chose to use a BP or when she started to use a BP. However, high PPBMI was associated (P < 0.03) with an earlier last use of a BP. Among women with a high PPBMI, those who used a BP breastfed for a longer period (P < 0.001) and ceased BF for different reasons than those who did not use a BP. Among women with a high PPBMI who used a BP, those who were unsuccessful at expressing their milk experienced several difficulties with BF and stopped BF sooner (P = 0.0003) than those who were successful at expressing their milk. Thus, high PPBMI was not a barrier to use of a BP and those who chose to use one were able to maintain dyadic BF for a longer period. (Funded in part by USDA Hatch grant NYC399427 and FDA.)
Protein quaternary structure focus for small molecule allosteric control
Authors: Jaffe Eileen K , Lawrence Sarah H , Selwood Trevor
Published: June 2021

A subset of homo‐oligomeric proteins can reversibly come apart, change conformation in the dissociated state, and reassemble into a structurally and functionally distinct oligomer. Work with the enzyme porphobilinogen synthase has established that this sort of quaternary structure dynamic is physiologically relevant. Proteins that can rearrange in this way are called morpheeins and their quaternary structure dynamic impacts our understanding of protein function and small molecule protein interactions in the following ways. 1) The quaternary structure dynamic of morpheeins can form a structural basis for allostery. 2) Small molecule stabilization of one of the alternate quaternary assemblies defines a new approach to allosteric drug discovery. 3) In addition to functioning as drugs, small molecules that bind to oligomer specific surface cavities will alter the quaternary structure equilibrium and can cause off‐target drug side effects. 4) Naturally occurring mutations can alter the normal quaternary structure equilibrium. This alters normal function and can contribute to disease states. We describe how each of these four impacts has been demonstrated with the morpheein porphobilinogen synthase. We propose additional medically relevant proteins whose behavior suggests that they may function as morpheeins.
Exploring Hydra Biology Using Chemical Genetics
Authors: Glauber Kristine Marie , Dana Catherine E. , Park Steve S. , Li Tao , Chamberlin A. Richard , Steele Robert E.
Published: June 2021

Studies using chemical genetics allow the researcher to perform the equivalent of classical genetics in real time using molecules that can produce a phenotype when applied to either cultured cells or a whole organism. We have initiated a chemical genetic screen using the AEP strain of Hydra vulgaris to identify bioactive molecules that modulate signaling pathways involved in development and regeneration. One compound, DAC‐2–25, induced the growth of ectopic tentacles in regenerating heads and buds. Chronic exposure to DAC‐2–25 induces ectopic tentacles in H. vulgaris AEP, H. vulgaris 950f, and H. viridissima, but not H. vulgaris Zurich. In chronically treated animals, the first ectopic tentacles appear just below the tentacle ring. Subsequent tentacles emerge in a wave down the body column. We are using AEP/Zurich chimeras to identify which cell lineage is targeted by DAC‐2–25. Structure‐activity relationship (SAR) studies have been initiated to determine what features of the molecule are required for activity. Ultimately we will use affinity chromatography coupled with mass spectrometry to identify the protein target of DAC‐2–25. Crosses of AEP and Zurich have been produced with the intention of doing bulk segregant analysis as an alternative approach to identifying the gene encoding the DAC‐2–25 target protein.
Acculturation factors and usual folate intakes among Mexican American women: National Health and Nutrition Examination Survey (NHANES), 2003–2006
Authors: Hamner Heather Carter , Cogswell Mary E , Johnson Mary Ann
Published: June 2021

MA women are at a higher risk for having a pregnancy affected by a neural tube defect which could be prevented by adequate folic acid consumption; however, research has suggested the acculturation could impact dietary intakes. We compared usual folate intakes in non‐Hispanic white (NHW) (n = 982) and Mexican American (MA) (n = 651) women aged 15–44 years by acculturation factors. We estimated usual folate intakes accounting for within‐individual variance using two 24 hour diet recalls and average daily supplement intake from NHANES 2003–2006. Median usual folic acid intakes from fortified foods alone did not differ between NHW and MA women, 173 and 183 ug/d, respectively. Median usual intake of natural food folate did not differ between NHW and MA women, 170 and 182 ug/d, respectively. Compared to NHW women, MA women had lower median usual total folic acid intakes (275 vs. 229 ug/d) and were less likely to report use of supplements with folic acid, 37% (95% confidence interval [CI]: 31%, 57%) vs. 20% (95% CI:15%, 25%). The reported use of supplements with folic acid was 34% (95% CI: 23%, 48%), 12% (95% CI: 4%, 28%), and 13% (95% CI: 7%, 23%) among MA women who reported speaking English, speaking English and Spanish, or speaking Spanish, respectively. Programs aimed at increasing folate intake should focus on reaching women who are less likely to use supplements with folic acid such as MA women who speak Spanish. Research funded by CDC.
CHARACTERIZATION OF SELENOPROTEIN V
Authors: Moustafa Mohamed El‐Sayed , El‐Srouji Rowan
Published: June 2021

Selenoprotein V (SelV) is a mammalian selenoprotein encoded on chromosome 19. SelV is a globular protein that occurs in the seminiferous tubules of the testes. Its function is yet unknown. We used bioinformatics programs to study structure‐function relationship of SelV. Protein calculator program revealed that the molecular weight of SelV in human is about 37 kDa. Human SelV is a basic protein with isoelectric point of 9.65. Using bioinformatics programs, we predicted that SelV contains 5 exons, where the selenocysteine (sec) residue was predicted to be in exon 2. SECISearch 2.19 program revealed that the selenocysteine insertion sequence (SECIS) element belongs to type I structure. We also predicted that SelV contains three domains: The N‐terminal domain is proline‐rich; the second domain is the hydrophobic domain containing 20 amino acid residues as well as the selenocysteine residue; and the C‐terminal domain which is rich in glutamate and lysine. Further studies are required to understand why mammalian SelV is rich in proline residues and why selenocysteine residue resides in hydrophobic domain in this selenoprotein.
Analysis of selenium in meats and meat products using INAA‐method
Authors: Lee Okhee , Chung Yongsam , Moon Jongwha
Published: June 2021

There are little reports on the Se nutritional status of Koreans because of deficient Se database on foods. Furthermore, accuracy of existing data is questionable because they were analysed by AAS or ICP methods which have large losses of Se during pre‐treatment process. This study was aimed to evaluate the Se contents in meat and meat products using INAA‐method. The Korean preferred meats and meat products were selected. To minimize contamination, all the instruments and storage tools were used after washing with HNO3. The homogenized food samples of 10~20g were freeze‐dried at temperature of − 60 C for 48~96 h. Subsequently, instrumental neutron activation analysis was employed to determine the contents of Se in the freeze‐dried food samples. Analytical quality control was conducted using NIST SRMs. The analysis limit of Se in foods by INAA was 0.01ppm. Se contents in meat and meat products were in the range of 100ug~0.01ug. These results showed that selenium contents in meat and meat products are varied. These results could be used to accumulate basic data for Se contents in Korean foods and evaluate the nutritional status of Korean
Urine potassium is a valid biomarker for diet potassium and a surrogate measure of compliance to a high dairy intervention in teenage girls
Authors: Rafferty Karen , Lappe Joan
Published: June 2021

Dietary intake records are used in nutrition intervention studies not only as a tool to assess reported intakes, but as a measure of compliance to a planned intervention. In this crossover study we assessed the validity of urinary potassium as a recovery biomarker and surrogate indicator of compliance to a dairy milk intervention. Ten healthy teenage girls 13–18 years old (mean 15.2) were assigned to consume a nutritionally adequate diet controlled for potassium and calcium, for four days. The study included one neutral control (water) and a test substance (milk and/or yogurt.). Each subject was studied twice and served as her own control. The control and test substances were consumed as part of pre‐selected meals for 2 consecutive 24‐hours periods one week apart. The control meal (with water) provided approximately 1500 mg potassium. The test meal (with dairy) provided approximately 3000 mg potassium. Consecutive 24‐hour urines were collected during the dietary tests. Previously we have shown correlations between diet potassium and urinary potassium in adult women studied under metabolic conditions, with a urineK:dietK ratio of 0.92. In the present study the mean 24‐hr. urine K was 1071 mg/d on the low potassium (control) diet, with a urineK:dietK ratio diet of 0.776. On the high potassium (dairy) diet the mean 24‐hr. urine K was 1742 and the urineK:dietK ratio was 0.664.
Vitamin D Status in Saudi Arabian Women: Is there an Association with Breast Cancer Risk?
Authors: Yousef Fatimah Mohammadali , Thomson Cynthia , Kumosani Taha , Yousef Jehad
Published: June 2021

The role of vitamin D in breast cancer prevention is an area of active research and to date the limited evidence suggests an inverse association. Saudi women are thought to be at greater risk for vitamin D deficiency because of their skin type and reduced UV exposure. Yet, there is a lack of data regarding the vitamin D status of Saudi women. The purpose of this research is to describe vitamin D status in a de‐identified sample of women in Saudia Arabia, a data set being collected in the context of a case‐control study to compare vitamin D status in women with and without a diagnosis of breast cancer.
Differential Expression of Carotenoid Metabolism Related Genes in Human Lung Epithelial Cells
Authors: Gong Xiaoming , Marisiddaiah Raju , Wiener Doris , Rubin Lewis
Published: June 2021

Epidemiologic studies have demonstrated that dietary beta‐carotene and total carotenoids are inversely associated with non‐small cell lung cancer risk. While high‐dose beta‐carotene supplementation for chronic smokers has been linked to increased lung cancer risk in clinical trials, little is known about the carotenoid metabolomic alterations that characterize neoplastic progression. Using two human pulmonary cell lines, we determined the expression of genes regulating carotenoid metabolism in normal bronchial epithelial (BEAS‐2B) and lung adenocarcinoma pulmonary epithelial (A549) cells. We assayed gene pathways including ALDH1A2, ALDH8A1, CMO1, CMO2, Cyp26A1, Cyp26B1, Cyp26C1, LRAT, RHD5, RHD8, RDH11 ~ 14, RHD16 and SDR16C5 by qRT‐PCR. Our results indicate that carotenoid metabolism‐related genes are expressed as different and distinct patterns in BEAS‐2B and A549 cells. BEAS‐2B cells express high levels of CMO2 and LRAT but not CMO1, Cyp26A1 or Cyp26B1, indicating an active lycopene metabolic apparatus. In contrast, A549 cells highly express CMO1, ALDH1A2, Cyp26A1 and Cyp26B1. In A549 cells, we could detect only low levels of CMO2 and no LRAT expression, implying presence of active beta‐carotene metabolism. We further tested this hypothesis by treating BEAS‐2B and A549 cells with lycopene. CMO2 and LRAT expression were increased but there were no effects on CMO1 expression. Finally, immunohistochemistry of NSCLC tissues shows profuse CMO1 staining in the tumors but little in adjacent normal lung. These findings suggest that specific beneficial effects of carotenoid on lung health may be linked to CMO2‐dependence in tissue. A shift to decreased expression of CMO2 and LRAT and increased CMO1 and Cyp26 family genes can alter carotene metabolism may contribute to lung carcinogenesis. (NIH HD42174, Muma Family Endowment)
Racial differences in correlations between reported dietary carotenoid intakes and plasma carotenoid levels
Authors: Arab Lenore , Cambou Mary Catherine , Craft Neal , Wessling‐Perry Katherine , Jardack Patricia
Published: June 2021

Because of cost and practicality, a dietary assessment tool that can accurately predict circulating plasma carotenoid levels is of great interest to epidemiologists. However, validity across racial groups needs to be assured. We studied dietary and plasma levels of oxycarotenoid levels (lutein & zeaxanthin, α‐, β‐cryptoxanthin) and hydrocarbons (lycopene, α‐, β‐carotene) among 123 healthy, non‐smoking adult African‐American (AA) and 128 Caucasian (CA) participants from Los Angeles. Age and gender adjusted correlations between diet and plasma differed substantially by race as seen in the Table. Despite similar plasma carotenoid levels, the correlations with dietary intakes were 0.09 to 0.40 lower among AAs. The 6‐day mean intakes from 24‐hr recalls for all carotenoids were lower among the AAs, consistent with similar validation studies. These results suggest that either this dietary assessment tool was less valid among African‐Americans or that other factors influencing circulating carotenoid levels differ between these groups. Supported by NIH R01CA105048.

Pearson's Correlations: Diet and Plasma Carotenoids

Carotenoid
AA N = 123
CA N = 128
Difference

Oxy Carotenoids

Lutein and Zeaxanathin
0.11
0.51
0.40

Total‐Cryptoxanthin
0.29
0.50
0.21

β‐Cryptoxanthin
0.30
0.50
0.20

α‐Cryptoxanthin
0.12
0.31
0.19

Hydrocarbons

Lycopene
0.00
0.09
0.09

β‐Carotene
0.22
0.41
0.19

α‐Carotene
0.18
0.38
0.20
Effect on Anti‐Diabetic Effect of the Mushrooms Grown in Germinated Brown Rice Extracts in Rin and 3T3‐L1 cells
Authors: Cha Youn‐Soo , Park Jeong‐Eun , Kang Hyun‐Ju , Soh Ju‐Ryoun , Beak Sung‐Hyun
Published: June 2021

This study investigated the anti‐diabetic activity of functional mushrooms grown in germinated brown rice (FMBL). FMBL extracts were obtained by using methanol extraction and concentration processes. First, insulin secretion‐stimulatory activity of FMBL was examined using the RIN cell line. Insulin and glucose secretion was enhanced in the FMBL extract treatment groups as compared to the non‐treated cells. In addition, the anti‐diabetic effects of FMBL extracts were confirmed by its effects on insulin levels, triglyceride content and PPAR‐γ mRNA expression, and insulin sensitivity in 3T3‐L1 cells. FMBL extract also reduced the PPAR‐γ mRNA expression and triglyceride accumulation in a dose dependent manner as compared to the control. The present study suggests that FMBL deserves to be evaluated for in vivo anti‐diabetic activity in animal and human studies.
Morinda Citrifolia (Noni) Inhibits Hepatic Gluconeogenesis via Increased Phosphorylation of Forkhead Transcription Factor (FoxO1)
Authors: Nerurkar Pratibha Vivek , Nishioka Adrienne , Eck Philip O , Johns Lisa M , Volper Esther , Nerurkar Vivek R.
Published: June 2021

Renewed interest in traditional medicine prompted us to investigate anti‐diabetic effects of Hawaiian medicinal plant, Morinda Citrifolia (noni) in mice fed high‐fat‐diet (HFD). Preliminary studies indicate that noni juice (NJ) improved glucose metabolism in mice fed HFD. Glucose production is increased in diabetes due to the inability of insulin to suppress gluconeogenesis. Insulin inhibits gluconeogenesis by modulating transcription factors such as forkhead box O (FoxO1). Since recent studies indicate that NJ synergistically augment insulin action in diabetic rats, we tested the hypothesis that NJ improves glucose metabolism by modulating gluconeogenesis via FoxO1 phosphorylation. In our studies hypoglycemic properties of NJ were associated with inhibition of hepatic FoxO1 mRNA expression and protein levels with a concomitant increase in FoxO1 phosphorylation and nuclear expulsion. Gluconeogenic genes, phosphoenolpyruvate C kinase (PEPCK) and glucose‐6‐phosphatase (G6P) were also significantly inhibited in mice fed HFD with NJ. Specificity of NJ‐associated FoxO1 regulation of gluconeogenesis was confirmed by siRNA studies using human hepatoma cells, HepG2. Our data suggests that NJ can offer an economical alternative, specifically for culturally sensitive diabetic individuals or those who cannot afford conventional medicine. [USDA‐CREES, 2004‐34135‐15182; NCCAM, R21AT003719].
Effects of regular black tea consumption on the antioxidant activity
Authors: Fisunoglu Mehmet , Besler Halit Tanju
Published: June 2021

Tea consumption is associated with the reduced risk of coronary heart disease.

In order to evaluate the effects of black tea consumption on the antioxidant activity, we designed a randomized, controlled study that involves 5 servings (200 ml)/d of black tea for 6 weeks in 16 healthy female volunteers who do not normally consume tea in their life. Subjects were randomly divided into 2 groups (study and control). Plasma levels of total antioxidant activity (TOAC), malondialdehyde (MDA), oxidized LDL (ox‐LDL), 8‐isoprostane (8‐epi PGF2α), and some amino acids susceptible to oxidation, namely histidine, proline, lysine, threonine, leucine, isoleucine, tyrosine, phenylalanine, tryptophan, and valine were measured.

TOAC significantly increased in the study group compared to the control group (1.81 ±0.1 vs 1.68 ±0.1). Plasma 8‐epi PGF2α level significantly decreased in the study group compared with the baseline (181.4 ±85.9 vs 164.0 ±94.1) however did not statistically changed between two groups. Although there was a positive trend in the results of oxidative biomarkers (MDA 1.46± 0.14 vs 1.42± 0.50, ox‐LDL 1928.9± 373.9 vs 1892.6± 284.6), we did not find significant difference between two groups. Amino acids susceptible to oxidation, except tryptophan, were significantly increased (p< 0.05) in the study group.

Daily 5 servings of black tea consumption significantly increased antioxidant activity.
Antidiabetic Effect of Color Potatoes in Streptozocin‐Induced Diabetic Rats
Authors: Kang Myung‐Hwa , Choi Mi‐Kyong , Jeon Ye‐Sook
Published: June 2021

The purpose of this study is to investigate the antidiabetic effect of color potatoes in streptozotocin‐induced diabetic rats. Total 49 Sprague‐Dawley rats were divided into 7 groups according to kinds of potatoes and treatment levels, that is, control, STZ, STZ with 10% color potato, STZ with 20% color potato, STZ with 10% potato, STZ with 20% potato, STZ with potato pigment. 10% potato, 20% color potato and potato pigment showed blood glucose lowering effect in glucose tolerance test. In the blood glucose change, 10% or 20% color potatoes significantly decreased blood glucose with dose‐response in 7 weeks and potato pigment significantly decreased blood glucose. While blood creatinine and BUN significantly lower, blood insulin was higher than those of STZ group. In pathological biopsy of kidney and pancreas, 20% color potato and potato pigment relieved indisposition views such as tubular degeneration of cortex, tubular mineralization of medulla in kidney, and islet cell atrophy in pancreas. These results of this study support that the treatment of 20% color potato and potato pigment have antidiabetic effect in improving blood glucose decrement in diabetic rats.
Resistant starch content of rice varies with rice variety but not cooking method
Authors: Stewart Maria L , Manglicmot Leilani
Published: June 2021

Rice is a staple carbohydrate in much of the world, including Hawaii. Previous work indicated that resistant starch (RS) content of rice consumed in India varied with rice variety and cooking method. This study aimed to quantify RS in 4 rice varieties commonly consumed in Hawaii (jasmine, long grain, medium grain, and short grain) cooked in three manners (baked, conventional rice cooker, and pressure cooker). Resistant starch in fresh and freeze‐dried rice was measured using AOAC method 2002.02 (n=2). ANOVA and paired t‐test were used to determine significant differences. Within each rice variety, cooking method did not affect RS content. However, within the baked method, jasmine rice had significantly more RS than medium and short grain rice. When cooked in a conventional rice cooker, jasmine rice and long grain rice had significantly more RS than short grain rice. Pressure cooked samples did not differ. Freeze‐dried samples reported significantly lower RS than fresh samples when corrected for original moisture content. In the state of Hawaii, diabetes is a major public health concern. This data will be used to evaluate glycemic response of selected rice varieties in healthy humans.
Regulation of ATP Binding Cassette Transporter A1 (ABCA1) and ABCG1 by Fatty Acids in RAW 264.7 Macrophages
Authors: Ku Chai Siah , Park Young‐Ki , Coleman Sara L , Seo Jeong Min , Lee Ji‐Young
Published: June 2021

As major transporters for cholesterol efflux in macrophages, ATP‐binding cassette transporter A1 (ABCA1) and ABCG1 play a pivotal role in reverse cholesterol transport, a major athero‐protective property of HDL. We investigated effect of fatty acids (FA) on ABCA1 and ABCG1 expression in RAW264.7 macrophages. Cells were treated with various FA, e.g., 16:0, 18:1, 18:2, 20:5 and 22:6 (100 μM) and T0901317, a liver X receptor (LXR) agonist. Realtime PCR revealed that unsaturated FA (UFA), not saturated FA, reduced basal expression of ABCA1 and ABCG1, which was abolished by T0901317. Trichostatin A, a histone deacetylase (HDAC) inhibitor, increased basal expression of ABC transporters and abrogated UFA‐induced repression. HDAC3 siRNA increased transporter mRNA by 3‐fold. Although repression of ABCA1 and ABCG1 mRNA by UFA was abolished by T0901317, protein levels remained diminished. When cells treated with 16:0 and 18:2 after knockdown of protein kinase C δ (PKCδ), reduction in ABCA1 and ABCG1 protein by 18:2 was further accentuated. Calpeptin or lactacystin, inhibitors for calpain and proteosome inhibitor, could not counteract 18:2‐induced reduction in protein. In conclusion, UFA repress ABCA1 and ABCG1 expression in macrophages by two mechanisms: LXR‐dependent transcriptional repression involving co‐repressor complex with HDAC3; calpain‐ and proteosome‐independent post‐transcriptional inhibition.

Grant Funding Source: Faculty Seed Grant from University of Nebraska‐Lincoln Research Council and Layman Award from University of Nebraska Foundation
Inhibitory Effect of a Mori Cortex Water Extract on Th2 Cytokine Expression and β‐Hexosaminidase Release by IgE Antigen Complex‐Stimulated RBL2H3 Cells
Authors: Kim Jeong Mi , Chung Mi Ja , Kim Dae Jung , Kim Tae Hyuk , Baek Jong Mi , Lee Sang cheol , Kim Cheon An , Choe Myeon
Published: June 2021

Mori Cortex is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of bronchial asthma but the underlying mechanisms involved are not fully understood. We investigated the antiasthma effects of a Mori Cortex water extract (MCWE) on the β‐hexosaminidase release and expression of interleukin (IL‐4), IL‐13 and tumor necrosis factor‐alpha (TNF‐α) mRNA and protein by IgE‐antigen complex‐stimulated RBL2H3 cells. Stimulation with the IgE‐antigen‐complex induced the release of β‐hexosaminidase and expression of IL‐4, IL‐13 and TNF‐α mRNA and protein. MCWE inhibited IL‐4 and IL‐13 cytokine transcription and translation against stimulation with the IgE‐antigen‐complex and MCWE also inhibited the β‐hexosaminidase release. Thus, MCWE might suppress the Th2‐mediated response through the down regulation of the production of Th2 cytokine and inhibition of degranulation.(This work is Supported for Corporate‐affiliated Research Institute of Academic‐Industrial‐Institutional Cooperation Establishment Business.)
Water soluble extracts from multiple mushroom varieties inhibit activation of Jurkat cells
Authors: Metzger Brandon , Williams Melissa G , Barnes David M , Hanlon Paul R
Published: June 2021

Mushrooms have traditionally been used for the modulation of immune function. Hot water, ethyl acetate and hexane extracts from Coriolus versicolor (turkey tail), Hericium erinaceus (bearded tooth), Ganoderma lucidum (reishi), Grifola frondosa (maitake), Lentinula edodes (shiitake), and Cordyceps sinensis (caterpillar fungus) were compared for their ability to reduce T lymphocyte activation. Jurkat immortalized T lymphocytes were pretreated with 1, 5 and 7.5 mg of mushroom extract per mL of media for 1 hour prior to activation by phytohemagglutinin (PHA) and phorbol 12‐myristate 13‐acetate (PMA). IL‐2 secretion by Jurkat cells was measured 24 hours after stimulation. Hot water extracts from all mushroom varieties reduced IL‐2 concentrations in a dose‐dependent manner, and extracts from Cordyceps sinensis and Grifola frondosa were the most potent. The ethyl acetate extracts from the mushrooms had similar effects as the hot water extracts, although in general they were less potent. Hexane extracts from all mushroom varieties had the least effect on IL‐2 production. Together these data demonstrate that water soluble compounds in the mushrooms tested have the greatest anti‐inflammatory bioactivity.
Role of mTOR/p70S6K in amino acid induced cardiac protection
Authors: Katayama Erika , Tsutsumi Rie , Nakaya Yutaka , Hirose Kayo , Tanaka Katsuya , Oshita Shuzo , Tsutsumi Yasuo M.
Published: June 2021

Leucine, as an essential amino acids and activator of mammalian target of rapamycin (mTOR), promotes protein synthesis and suppresses protein catabolism. Recent studies have showed that the mTOR/p70S6K complex may mediate the beneficial effects to ischemia‐reperfusion injury. We therefore proposed that amino acids may produce cardiac protection against myocardial ischemia‐reperfusion mediated by mTOR/p70S6K.

Mice were subjected to 30 min of myocardial ischemia followed by 120 min of reperfusion. Control mice were exposed to ischemia and reperfusion only. Other mice were randomly assigned to receive amino acids, administered 15 min before ischemia and/or rapamycin, a mTOR inhibitor, were given 15 min prior to amino acids preconditioned.

Amino acids preconditioned mice were significantly reduced infarct size compared to control group (34 ± 4% vs. 45 ± 3% [n = 3] respectively, p<0.05) and rapamycin eliminated the protection produced by amino acids (41 ± 3% [n = 3]). Additionally, the hearts treated with amino acids had a 2‐fold increase in phosphorylation of both mTOR and p70S6K, and pretreated with rapamycin completely abolished mTOR/p70S6K phosphorylations in response to amino acids treatment.

These results indicate that amino acids induced myocardial protection involves activation of the p70S6K that is mediated by the mTOR.
Using whole‐body phenylalanine kinetics to measure the effects of amino acid supplementation in mature horses
Authors: Urschel Kristine L. , Geor Raymond J. , Wagner Ashley L. , Williamson Lindsey M. , Smith Tracy L. , Hanigan Mark D. , Harris Pat A.
Published: June 2021

Stable isotopes have not been used in horses to measure whole‐body protein kinetics or to assess the adequacy of a diet's amino acid profile. The objective of this study was to determine whether supplementing a control diet (CON) with equal (0.38 mmol/kg/d) amounts of either glutamate (+GLU), leucine (+LEU), lysine (+LYS) or phenylalanine (+PHE) would affect whole‐body PHE kinetics in mature horses. 12 mature male horses were each studied 3 times: once while receiving the CON diet (n=12) and while receiving 2 of the 4 supplemented diets (n=6/diet). Following 7d of adaptation, horses received two 6h primed, constant infusions each period: [1‐13C]PHE (d8) and [13C]bicarbonate (d9). Breath samples were collected to measure the rate of 13CO2 appearance, to calculate PHE oxidation (d8) and total CO2 production (d9). Blood was sampled (d8) to calculate PHE flux and kinetics using a stochastic model. Horses receiving the +LEU, +LYS and +PHE diets had higher plasma concentrations of their supplemented amino acid than when they received the CON diet (P < 0.05). Relative to CON, horses receiving +PHE had a greater PHE flux (+20%) and rate of PHE oxidation (+111%) (P < 0.05). There were no differences in PHE use for protein synthesis relative to CON for any of the supplemented diets (P > 0.05). The isotope methodology was sensitive enough to detect differences in PHE oxidation between diets. Amino acid supplementation of the CON diet did not appear to increase whole‐body protein synthesis in mature horses. Funded by WALTHAM Centre for Pet Nutrition.
Phosphatidylinositol 3‐Kinase mediates the ability of retinol to inhibit colon cancer cell invasion in vitro
Authors: Griffin Jennifer N , Lane Michelle A
Published: June 2021

Vitamin A supplementation decreases colorectal cancer metastasis and retinol inhibits phosphatidylinositol 3‐kinase (PI3K) activity. Our objective was to determine if PI3K mediates the effects of retinol on colon cancer invasion in vitro. HCT‐116 cells expressing wild‐type or constitutively active (ca) PI3K were treated with 0 (vehicle control) or 10 μM retinol for 24 hours. Matrigel‐coated Boyden chambers measured invasion. Western blot analyses determined β‐catenin, matrixmetalloproteinase (MMP)‐9 and TIMP (tissue inhibitor of MMP)‐1 levels. Zymography detected MMP‐9 activity. Our results show caPI3K blocked the ability of retinol to decrease invasion, β‐catenin and MMP‐9 protein, MMP‐9 activity, and increase TIMP‐1. Specifically, 10 μM of retinol resulted in cell invasion of 103.1 ± 6.7 and 65.9 ± 5.5, β‐catenin levels of 126.8 ± 15.1 and 63.5 ± 17.4, MMP‐9 levels of 96.2 ± 2.9 and 80.8 ± 6.2, TIMP‐1 levels of 81.7 ± 29.9 and 121.7 ± 8.9, and MMP‐9 activity of 81.5 ± 19.7 and 73.3 ± 4.7% vehicle control ± SEM for the caPI3K and wild‐type PI3K cell lines, respectively. Thus, PI3K mediates the ability of retinol to decrease colon cancer invasion. Sponsor: NIH grant #5 R21 CA120414‐02.

Grant Funding Source: NIH grant #5 R21 CA120414‐02
Justicidin A inhibits AKT/mTOR and activates type III PI3K/beclin 1 signaling pathways leading to autophagy of human colorectal cancer cells
Authors: Su Chun‐Li , Yen Cheng‐Hsin , Lin Chun‐Nan , Won Shen‐Jeu
Published: June 2021

Justicidin A (JA), a pure compound isolated from Justicia procumbens, is a traditional herbal remedy for treatment of fever, pain, and cancer in Taiwan. Treatment of JA not only increased the expression of autophagic marker microtubule‐associated protein 1 light chain‐II by immunoblotting and confocal microscopy, but also augmented the formation of acidic vesicular organelles by flow cytometry in HT‐29 cells. Further study indicated that the expression of p‐AKT, p‐mTOR, p‐p70S6k was decreased whereas that of type III PI3K, becline 1, and Atg5‐Arg12 was increased in JA‐treated cells. Annexin V‐FITC and propidium iodine staining followed by flow cytometry revealed that induction of early apoptosis was displayed at 12 h that was later than the induction of autophagy. Of note, pre‐treatment of the cells with autophagy inhibitor 3‐methyladenine reduced JA‐induced early apoptotic rate. Oral administration of JA (6.2 mg/kg) once a day for 56 consecutive days was shown to suppress the growth and induce apoptosis and autophagy of HT‐29 cells transplanted to NOD‐SCID mice, suggesting chemotherapeutic potential of JA on human colorectal cancer cells.

(Supported in part by National Science Council, Taipei, Taiwan, No. NSC 96‐2313‐B‐309‐001‐MY2 and NSC 98‐2313‐B‐003‐002‐MY3).
Oral amino acids and testosterone therapy to improve muscle mass and quality of life during chemotherapy in recurrent cervical cancer: A case study
Authors: Dillon Edgar Lichar , Randolph Kathleen M , Casperson Shanon L , Durham William J , Urban Randall J , Levine Lyuba , Diaz‐Arrastia Concepcion , Sheffield‐Moore Melinda
Published: June 2021

Muscle wasting is by far the most important phenotypic feature of cancer cachexia, though little success has been achieved toward reversing or preventing this catabolic process. Anabolic interventions including testosterone and amino acid supplements may be beneficial in reducing and/or reversing muscle wasting in these patient populations.

A 48‐year‐old recurrent cervical cancer patient was scheduled to receive three 21‐day cycles of cisplatin and topotecan chemotherapy. She consented to a 10 week blinded interventional pilot study were she received daily amino acid supplements and weekly injections of testosterone enanthate before and during the weeks of chemotherapy treatment. Blood, Dual Energy X‐ray Absorptiometry (DEXA) scans, and Functional Assessment of Cancer Therapy ‐ General (FACT‐G) questionnaire data were collected before and after the 10 week period.

Total serum testosterone concentrations, total and trunk lean body mass, and body weight increased after 10 weeks. Total and trunk fat mass decreased. FACT‐G scores improved.

Concomitant treatment of oral amino acids and testosterone may be a viable therapeutic option for fighting cachexia and improving quality of life during chemotherapeutic treatment of recurrent cervical cancer. A double‐blinded, placebo‐controlled interventional study is in progress (NCT00878995).

Funding through NIH/NCI RO1 CA127971 and T32 CA117834.
Association between Serum Cytokine Concentrations, Flavonol Intake and Colorectal Adenoma Recurrence in the Polyp Prevention Trial (PPT)
Authors: Bobe Gerd , Murphy Gwen , Albert Paul S. , Sansbury Leah B. , Schatzkin Arthur , Lanza Elaine , Colburn Nancy H. , Cross Amanda J.
Published: June 2021

Background

Serum cytokine concentrations are considered to be markers of inflammation and may be used to identify individuals at increased risk or in early stages of colorectal carcinogenesis. There are certain dietary components that may have anti‐inflammatory properties, including flavonols, which are found in fruits and vegetables.

Aims

To determine whether serum concentrations of IL‐1β, IL‐2, IL‐8, IL‐10, IL‐12p70, GMCSF, IFNγ, and TNFα, measured over time, are associated with flavonol intake and colorectal adenoma recurrence.

Methods

We estimated odds ratios (ORs) and 95% confidence intervals (CIs) for cytokine concentrations, flavonol intake, and adenoma recurrence in 872 participants from the intervention arm of the PPT.

Results

Flavonol intake was inversely related with IFNγ concentrations. A decrease in IL‐2 concentrations during the trial was associated with any adenoma recurrence (4th versus 1st quartile, OR = 1.68, 95% CI: 1.13–2.49) and advanced adenoma recurrence was inversely related with a decrease in IL‐1β (OR = 0.37, 95% CI: 0.15–0.94) and IL‐10 (OR = 0.39, 95% CI: 0.15–0.98). Individuals with above median flavonol intake plus equal or below median cytokine concentrations had the lowest risk of advanced adenoma recurrence.

Conclusion

High flavonol intake combined with a decrease in or low cytokine concentrations may be most protective against advanced adenoma recurrence.
Protein supplementation during 6 mo weight loss enhances body composition changes in older women
Authors: Mojtahedi Mina C , Thorpe Matthew P , Richey Abby L , Johnson Curtis L , Karampinos Dimitrios C , Georgiadis John G , Layman Donald K , Evans Ellen M
Published: June 2021

Adverse increases in adiposity and reductions in fat free mass (FFM) are age related changes in body composition and are linked to declines in physical function in older women. Higher protein intake in weight loss may augment fat mass (FM) loss and attenuate FFM loss, which may benefit function. This study assessed effects of 6 mo weight loss supplemented with powdered protein or placebo on body composition and function in older obese women. Women (63.8±3.4 yrs, BMI 32.9±4.1 kg/m2) were prescribed an energy‐restricted diet (1500 kcal/d; 15, 65, 30% of energy from protein, CHO and fat, respectively) and randomly assigned to 2 × 25g/d of whey protein (PRO, n=8) or maltodextrin (CARB, n=7) supplementation. Measures included whole body FM and FFM with DXA; thigh muscle, subcutaneous (SUBQ), intermuscular and intramyocellular fat with MRI; knee extension and flexion strength with isokinetic dynamometry. PRO trended towards greater weight loss (−10.6±6.7%, −5.0±4.0%; p=0.078), had greater reductions in FM (−18.9±10.6%, −8.0±4.2%; p=0.024) than CARB, but no significant difference in FFM (−4.0±4.9%, −2.0±4.1%; p=0.414). %SUBQ thigh fat change (−6.1±3.0%, −1.3±1.7%; p=0.003) was the only significant MRI outcome. Groups did not differ in leg strength changes. Higher protein intake in weight loss in older women enhances FM reductions while preserving FFM; however, these changes do not impact changes in physical function.

Grant Funding Source: National Dairy Council, Carraway Foundation