bluuebunny/crossref_metadata_2025_split huggingface dataset:
Arginine metabolism and sensing by mTOR in T cells
Authors: Senkevitch Emilee , Frauwirth Kenneth , Tuchman Mendel , Bequette Brian 
Published: June 2021

Arginine has been shown to play an essential role in T cells, as arginine depletion in the immune microenvironment has a negative effect on T cell function. The goal of this study was to characterize how T cells sense and metabolize arginine.Experiments were carried out using murine T cells stimulated by CD3 and CD28 as well as the lymphoma cell line EL4. Proliferation and IL‐2 ELISA was used to assess T cell function. Western Blots were performed to determine amino acid sensing by the mTOR pathway. To establish pathways of arginine metabolism, a stable arginine isotope was used and metabolic products were analyzed using GC‐MS. Metabolomic analysis by GC‐MS was used to identify any unsuspected signaling and metabolic pathways regulated by arginine. RT‐PCR was used to analyze expression of enzymes known to be involved in arginine metabolism.T cells stimulated in the absence of arginine fail to proliferate or increase production of IL‐2, and activated T cells increase their uptake of arginine. T cells differentially sense arginine, citrulline, and ornithine through mTOR. Urea cycle enzymes are not expressed in T cells. GC‐MS analysis showed that arginine is not a substrate for the Krebs cycle and potentially plays a role in fatty acid synthesis. This research was supported by a Howard Temin Award from NCI/NIH and a New Investigator Grant from the Leukemia Research Foundation.
Sequence Independent Phylogenetic Footprinting Analysis Identifies Novel Transcription Regulation for Orexin Receptors
Authors: Perez‐Leighton Claudio Esteban , Buterick Tammy Angeline , Billington Charles , Kotz Catherine 
Published: June 2021

The orexin receptors (OX1/2R) are G‐protein coupled receptors involved in the regulation of energy metabolism (Tsujino et al. 2009). Current evidence suggests that changes in expression of the OX1/2R are implicated in the development of obesity (Teske et al. 2006). However, there is limited information about the transcriptional regulation of these receptors. A sequence‐independent phylogenetic footprinting bioinformatic analysis of the OXR promoters from six species (human, rat, mouse, chimpanzee, dog and cow) identified several transcription factor binding sites (TFBS). Our preliminary results indicate the transcription factors (TFs) SP1, NFATc2, EGR1 and PPARγ are involved in the transcriptional regulation of OX1/2R. These TFs have been implicated in several rodent models of obesity, yet their specific role in the brain is undefined (Plagemann et al. 2009, van Beekum et al. 2009, T. C. Yang et al. 2006). This study is ongoing and we are currently analyzing TFBS co‐occurrences and OXR promoter's enhancer structures. Simultaneously, we have started working towards validation of the TFBS predictions in set of new immortalized rat hypothalamic cell lines, which either contain or do not contain both OXR. These findings will provide novel insight into the transcriptional regulation of the OX1/2R.Funding: The Department of Veterans Affairs and the National Institute of Health, NIDDK, DK078985‐02S2.
Improved purification scheme for Riboflavin Binding Protein
Authors: Milligan Michael , Turner Geraldine , Smith Sheila Rose , Benore Marilee Ann 
Published: June 2021

Riboflavin Binding Protein (RBP) is a glycophosphoprotein found in the eggs of oviparous species. Responsible for transporting riboflavin for the developing embryo,RBP also binds copper, suggesting other potential functions for this protein. Because of its stability and functional assets, RBP has been employed for use in structure‐activity studies and other uses. RBP is abundant in chicken egg albumen and yolk and can be isolated from eggs as well as from recombinant sources; several purification methods have been reported in the literature. Most procedures require one or more steps requiring high salt, low pH, or organic solvent extraction. Improved purification procedures that minimize stringent steps would allow for faster isolation as well as retention of all structural features. We have made improvements to the procedure for isolation of RBP, including modifications to the fractionation and chromatographic steps in the protocol.
The role of the alpha‐tocopherol transfer protein during zebrafish embryogenesis
Authors: Miller Galen William , Labut Edwin M , Lebold Katie M , Barton Carrie , Tanguay Robert L , Traber Maret G 
Published: June 2021

The alpha‐tocopherol transfer protein (TTP) is required for vitamin E bioavailability and transfer to the blood stream. The finding that TTP is expressed in mammalian uterine and placental cells emphasizes the significance of TTP during embryogenesis. To study TTP's role in fetal development, a non‐placental model is essential. The zebrafish meets these requirements. Expression of TTP mRNA in zebrafish embryos increases 10‐fold within the first 10 h of zebrafish development and increases again between 24 and 36 h post fertilization (hpf). TTP was knocked down using morpholinos (MO) designed to inhibit TTP mRNA processing by interfering with proper splicing or translational initiation. Attenuating TTP by blocking proper mRNA splicing in the zebrafish embryos caused a distinct “pinched” phenotype in >50% of the embryos, observed at 8 hpf, resulting from an irregular yolk sac engulfment, suggesting membrane instability. Corresponding with the second increase in TTP mRNA, both MOs at 24 hpf caused severe cell death in the developing brain (>50% and >95%, splice‐blocking and translation inhibition respectively) while concentration matched controls caused less (<5%). To control for non‐specific effects we injected a MO against p53, which resulted in less mortality by 8 hpf while preserving the TTP knockdown effects. Thus, TTP is vital for embryonic survival.Funding: NICHD 5R01HD062109‐02
Effects of Fasting on Food Choices
Authors: Tal Aner , Wansink Brian C. , Shimizu Mitsuru 
Published: June 2021

How does food‐deprivation effect food choices? The study reported here suggests that people who have suffered mild food deprivation lean more towards higher calorie foods and consequently consume more of them. Participants (N = 81) were instructed to avoid eating for 18 hours prior to participating in the study during lunch. Importantly, only participants who reported not normally skipping breakfast were chosen for the study, as it was hypothesized that the manipulation would be more efficacious for them. Since not all participants obeyed instructions we only counted those who skipped breakfast as our fasting condition. We examined the order in which they chose to eat 6 different foods belonging to 3 different food groups: carbohydrates (dinner rolls and fries), protein (chicken fingers and cheese), and vegetables (green beans and carrots). Analyses revealed that fasting participants started with carbohydrates first (41.03%) to considerably greater extent than did non‐fasting participants (20%): χ2= 4.37, p = .04. Carbs replaced vegetables as the first chosen food. Choice of starting with vegetables went down from 52.5% for control participants to 23.08% for fasting participants. Importantly, eating a food first led to consuming more of a food, r = .24 (p < .0001). Effects were particularly marked for females. No fundingGrant Funding Source: None
Cellular reprogramming by bacterial lipoprotein rescues ST2‐deficient mice from lethal sepsis via acceleration of phagosome maturation
Authors: Wang Jiang Huai , Buckley Julliette M. , Liu Jinghua , Blankson Siobhan , Di Wu Qiong , Redmond H. Paul 
Published: June 2021

BackgroundST2, a member of the Toll‐IL‐1 receptor (TIR) superfamily, negatively regulates both TLR2 and TLR4 signaling. However, it remains undefined whether or not ST2 influences phagocyte‐mediated antimicrobial responses.MethodsPolymicrobial sepsis was induced by cecal ligation and puncture in wild‐type and ST2‐deficient mice. Phagosome maturation was assessed by detecting phagosome acidification and phagosome/lysosome fusion.ResultsST2‐deficient mice were more susceptible to polymicrobial sepsis than their wild‐type littermates, with an increased production of proinflammatory cytokines. Bacterial clearance from the circulation and visceral organs following polymicrobial infection was significantly delayed in ST2‐deficient mice. This was associated with a reduced antimicrobial response and impaired phagosome maturation characterized in ST2‐deficient phagocytes. Cellular reprogramming by bacterial lipoprotein (BLP) tolerisation protected ST2‐deficient mice against polymicrobial sepsis‐related lethality and abrogated the proinflammatory effect of ST2‐deficiency. Moreover, bacterial clearance was accelerated in BLP‐tolerised, ST2‐deficient mice, with substantially enhanced bacterial uptake, phagocytosis and intracellular killing of either gram‐positive S. aureus or gram‐negative S. typhimurium, and accelerated phagosome maturation.ConclusionST2‐deficient mice exhibit an increased susceptibility to polymicrobial infection, and cellular reprogramming by BLP rescues ST2‐deficient mice from lethal sepsis by attenuating the overwhelming proinflammatory response, and simultaneously accelerating bacterial clearance via enhanced phagosome maturation and bactericidal activity.
Interleukin‐1‐induced cyclooxygenase‐2 and inducible nitric oxide synthase expression in human OA chondrocytes is associated with histone H3K4 methylation
Authors: El mansouri Fatima Ezzahra , Chabane Nadir , Zayed Nadia , Martel‐Pelletier Johanne , Pelletier Jean‐Pierre , Fahmi Hassan 
Published: June 2021

ObjectiveIn the present study, we investigated the role of H3K4 methylation in interleukin‐1β (IL‐1)‐induced COX‐2 and iNOS expression in human OA chondrocytes.MethodsChondrocytes were stimulated with IL‐1 for various time periods and the expression of iNOS and COX‐2 mRNAs and proteins were evaluated using RT‐PCR and Western blotting, respectively. H3K4 methylation at the iNOS and COX‐2 promoters was evaluated using ChIP assays. The role of histone methylation was further evaluated using the methyltransferase inhibitor, 5′‐deoxy‐5′(methylthio) adenosine (MTA).ResultsIL‐1 induced iNOS and COX‐2 mRNA and protein in a dose‐ and time‐dependent manner. The induction of iNOS and COX‐2 expression by IL‐1 was associated with H3K4 di‐ and trimethylation at the iNOS and COX‐2 promoters, whereas the levels of H3K4 monomethylation remained unchanged. Treatment with MTA inhibited IL‐1‐induced H3K4 methylation as well as IL‐1‐induced iNOS and COX‐2 expression.ConclusionThese results indicate that H3K4 methylation contributes to IL‐1‐induced iNOS and COX‐2 expression and suggest that this pathway could be a potential target for pharmacological intervention in the treatment of OA disease.
Absorption of zinc from mixed diets containing conventional or zinc‐biofortified Bangladeshi rice, or conventional Bangladeshi rice with added zinc, among young children in a peri‐urban community
Authors: Islam M Munirul , Woodhouse Leslie R , Hossain Mohammad Bakhtiar , Ahmed Tahmeed , Huda Nazmul , Ahmed Tanveer , Hotz Christine , Brown Kenneth H 
Published: June 2021

Rice cultivars with higher Zinc (Zn) contents have been produced through selective breeding, and in vivo Zn bioavailability information is needed.We measured fractional (FAZ) and total Zn absorption (TAZ) from mixed diets containing zinc‐biofortified rice (ZnBfR) compared with diets containing conventional rice (CR), with or without added Zn in children using a triple stable isotope technique. Healthy children (n=42; 36–59 mo) were recruited and divided equally into 2 groups. In a randomized 2 day cross‐over design, Group A received 150 g of ZnBfR (67Zn) for 1 day (4.97 mg total Zn intake), and 150 g CR (70Zn) on the other day (3.93 mg total Zn intake). Group B received 150 g of ZnBfR (67Zn) for 1 day, and 150 g CR (70Zn) with added ZnCl2 (CR+Zn) on the other day (5.85 mg total Zn intake). Mixed diets were served 3 times daily. 68Zn was infused intravenously at the end of day 2. Urine samples were collected on day 6–10, urinary Zn isolated and isotope ratios determined.Preliminary data indicate that FAZ (%) was 20.9a ± 4.1, 26.4b ± 4.2 and 20.2a ± 4.2 for ZnBfR, CR and CR+Zn (p <0.05). TAZ (mg) was 1.04a ± 0.21, 1.04ab ± 0.17 and 1.18b ± 0.24 for ZnBf, CR and CR+Zn diets (p <0.05). TAZ from a ZnBfR diet is as good as a diet based on CR but the amount of increased zinc intake from the ZnBfR rice may be insufficient to enhance TAZ compared with CR at the levels of consumption studied.
Trypanosoma cruzi regulates the defensin alpha‐1 interactome network and defensin alpha‐1 causes trypanosome membrane pore formation to control the early process of infection
Authors: Furtak Vyacheslav , Kleshchenko Yuliya , Nde Pius , Pratap Siddarth , Johnson Candice , Madison Nia , Lima Maria , Villalta Fernando 
Published: June 2021

The interactome networks of innate immune molecules operating in the early process of Trypanosoma cruzi infection of human cells are unknown. Here we show that T. cruzi regulates the defensin alpha‐1 interactome network and that human defensin alpha‐1 plays a fundamental role in the initiation of cellular responses to T. cruzi. We demonstrate that human epithelial cells respond to early T. cruzi infection by up‐regulating the expression and secretion of defensin alpha‐1, which causes trypanosome membrane pore formation and depolarization to inhibit T. cruzi motility which prevents cellular infection. The pores formed in the flagellar membrane of the invasive trypanosomes by defensin alpha‐1 cause the detachment of the flagellum from the trypanosome body to abolish trypanosome motility, which is required for trypanosome cell entry. This is the first report elucidating the defensin alpha‐1 interactome network and showing that defensin alpha‐1 disrupts the T. cruzi flagellum to compromise trypanosome motility and prevent cellular infection. Thus, defensin alpha‐1 possesses remarkably distinct modes of activity against protozoan parasites than bacteria and viruses, and its action may provide insights for the development of new anti‐trypanosomal strategies.This work was supported by NIH grants AI AI080580 and AI007281.
Food preparation methods among African American churchgoers in Baltimore, MD and their associations with body mass index (BMI) and food purchasing behaviors
Authors: Budd Nadine , Surkan Pamela , Martins Paula , Rowan Megan , Flamm Laura , Gittelsohn Joel 
Published: June 2021

Healthy Bodies Healthy Souls is a diabetes prevention program, which aims to increase physical activity, access to healthy foods, and healthy food preparation methods among urban African American (AA) churchgoers. Food purchasing and preparation methods were assessed among AA church members (n=136) from 6 churches in Baltimore City. Pan‐frying in fat was the most frequently used preparation method (17.6%). There was no correlation between BMI and unhealthy cooking methods (deep‐fried, pan‐fried in fat, broiled, baked, or grilled with fat) or healthy cooking methods (pan‐fried without fat, boiled or steamed without added fat, cooked with cooking spray, raw). There was a significant positive association between BMI and total preparation frequency of grouped foods (potatoes, leafy greens, chicken, ground beef, fish, eggs) (r =.173, p =.043) and with total beef preparation frequency (r = .224, p =.008). Total frequency of at‐home food preparation was positively correlated with frequency of unhealthy food purchasing (r =.402, p <0.001). Results suggest that the frequency of home food preparation and types of foods prepared, regardless of preparation methods used, play a role in determining BMI among this population. Efforts to encourage home meal preparation as a means of reducing the obesity epidemic must be accompanied by initiatives to improve cooking methods.Grant Funding Source: The American Diabetes Association
The effects of whey protein supplementation and exercise on bone in overweight/obese middle‐aged adults
Authors: McMorrow Aoibheann M , Weinheimer Eileen M , Conley Travis B , Kobza Vanessa M , Sands Laura P , Lim Eunjung , Janle Elsa M , Campbell Wayne W 
Published: June 2021

PurposeTo investigate the effects of whey protein (WP) supplementation and exercise training on bone mineral density (BMD) and content (BMC).MethodsIn this double‐blind, randomized, placebo‐controlled, community‐based 9‐month trial, 108 females (49±7 y, 30.1±2.9 kg/m2) and 78 males (49±8 y, 30.4±2.7 kg/m2) consumed 200 kcal supplements with 0 (n=68), 10 (n=71), 20 (n=21), or 30g (n=26) WP twice daily. All subjects performed resistance (2 d/wk) and aerobic (1 d/wk) exercise during the 9‐month intervention. DXA measurements were made pre, mid and post intervention.ResultsOver time (i.e. with exercise) total body and total femur BMD were unchanged. Femoral neck BMD decreased (−0.15±1.08%, P=0.044) independent of sex. Spine BMD decreased in women (−1.08±2.60%, P<0.001) and trended higher in men (0.52±2.83%, P=0.053) (sex × time P=0.001). In women, BMC decreased in the total body (−0.29±1.14%, P=0.001) and trended lower in the spine (−0.76±3.94%, P=0.074), while there were no significant changes in men (sex × time P=0.02). WP supplementation, regardless of dose, did not influence these responses (WP × time P>0.05).ConclusionWhey protein supplementation did not influence the bone mass of weight‐stable, middle‐aged overweight/obese men and women who completed a 9‐month exercise program.Support: US Whey Protein Research Consortium; NIH T32AG025671 and UL1 RRO25761, Purdue Ingest. Behav. Res. Ctr.
Role of Advanced Glycation End Products (AGEs) in Adipogenesis in Senescent Preadipocytes
Authors: Chen Chih‐Yu , Moon Yang Soo , Kim Kee‐Hong 
Published: June 2021

Obesity contributes to the development of metabolic syndromes and accelerates to onset of multiple aging‐related disorders. Adipogenesis is the main process to increase not only lipid accumulation, but also elevate inflammation and oxidative stress. Within aging, preadipocyte capacity to replicate and differentiate declines. Advanced glycation end products (AGEs), through its receptor (RAGE), have been shown to be implicated to aging‐associated chronic diseases. The inhibition of the AGE‐RAGE interaction has been proposed as a target to treat or prevent these diseases; however the role of AGE‐RAGE axis in obesity has not been delineated. The objective of this study was to characterize the function of AGE‐RAGE axis in adipogenic capacity of senescent 3T3‐L1 preadipocytes. We found that the high passage preadipocytes (HPP) (more than 15 passages) exhibited the elevated levels of replicative senescent markers, including upregulation of tumor suppressor protein 53, interleukin‐6 levels and beta‐galactosidase activity compared to low passage cells (equals to or less than 5 passages). Additionally, HPP had reduction of insulin sensitivity and adipogenic ability. Interestingly, HPP treated with AGEs restored adipogenic capacity assessed by Oil Red O staining and increased adipogenic gene expression such as peroxisome proliferator‐activated receptor gamma, CAATT/enhancer binding protein alpha, and fatty acid synthase. Moreover, AGE‐stimulated adipogenesis in HPP is related to increased RAGE protein expression. We further demonstrated that the blockage of RAGE by a neutralizing antibody blunted the pro‐adipogenic function of AGEs. Taken together, our results suggest that AGEs induce adipogenesis in a RAGE‐dependent manner in senescent preadipocytes.Grant Funding Source: Scholarship from the Ministry of Education of Taiwan
Tacrolimus and Rapamycin do not elevate cholesterol synthesis in post‐transplant patients, but may impair response to multi‐nutrient dietary intervention
Authors: Lambert Jennifer E , Ryan Edmond A , Thomson Alan BR , Clandinin Michael T 
Published: June 2021

Patients frequently experience hypercholesterolemia after transplant due to immunosuppressive drugs. Mechanisms responsible for this rise in plasma cholesterol are not clear. Dietary therapy is usually not sufficient to reduce plasma cholesterol post‐transplant, however these interventions may be limited in scope. The purpose of this study was to investigate cholesterol synthesis as a possible mechanism for hypercholesterolemia in post‐transplant patients, and test the efficacy of a multi‐faceted dietary intervention. Islet (ITx; n=8) and liver (LTx; n=7) post‐transplant patients on Tacrolimus (Tac) or Rapamycin (Rapa) were recruited with normal control subjects (Ctl; n=9) to measure 24h cholesterol fractional synthesis (FSR‐C) using deuterium. A subset of patients underwent a 4‐week dietary intervention using fish oil, phytosterols, soy, and fibers. FSR‐C was not different between Ctl, ITx and LTx groups. Diet intervention significantly lowered plasma cholesterol and triglyceride in Ctl, however was not effective in ITx or LTx. Diet produced mixed results in changes in FSR‐C across groups. In conclusion, hyperlipidemia caused by Tac or Rapa may not be due to elevated cholesterol synthesis. However, these drugs may interfere with normal responses to hypolipidemic dietary therapy effective in non‐transplant populations.Grant Funding Source: Canadian Institutes of Health Research
Impairment of endothelial function despite improvements in traditional lipid markers in low‐carbohydrate vs. high‐carbohydrate, high‐fiber diet in adults with metabolic syndrome
Authors: Gaesser Glenn , Angadi Siddhartha , Weltman Nathan , Rodriguez Jessica , Patrie James , Barrett Eugene , Brock David , Davis Christopher , Irving Brian , Weltman Arthur 
Published: June 2021

Twenty‐three adults (17 females) with the metabolic syndrome (IDF Criteria) participated in this randomized, single‐blind, crossover study to determine the effects of a low‐carbohydrate diet (LC: 15–20% carbohydrate (fiber ≤ 10 g/day), 55–60% fat, 25–30% protein) and a high‐carbohydrate, high‐fiber diet (HCHF: 55–60% carbohydrate (fiber ~45g/day), 20–25% fat,15–20% protein) on serum lipids and vascular endothelial function (brachial artery flow‐mediated dilation; BAFMD). Diets were ad libitum and lasted 4 weeks, with 4‐week washout. Significant decreases (p<0.05) in VLDL (−7.5 ± 10.6 mg/dl), triglycerides (−59.7 ± 73.2 mg/dl) and non‐HDL (−10.5 ± 22.7 mg/dl) were observed after LC while HCHF resulted in significant decreases in total Cholesterol (−13.2 ± 21.9 mg/dl), HDL (−3.9 ± 5.5 mg/dl), and non‐HDL (−9.2 ± 19.9 mg/dl). A strong trend for reduced BAFMD was observed after LC (10.1 ± 1.4% vs. 7.2 ± 1.1%; p=0.06), while BAFMD remained unchanged after HCHF (9.5 ± 1.3% vs. 10.4 ± 1.5%; p=0.61). A significant diet interaction was observed for changes in BAFMD (p=0.01). Despite improvements in fasting lipids following LC, endothelial function was impaired in comparison to the HCHF diet. Traditional fasting lipid markers may not accurately reflect the dysmetabolic setting that results in endothelial dysfunction following a LC diet in adults with metabolic syndrome.Funding: NIH RR MO100847; Wheat Foods Council