bluuebunny/crossref_metadata_2025 huggingface dataset:
Gut Commensal Immunomodulatory Factors: Identification and Structure‐Function Studies
Authors: Oh Sungwhan , Erturk‐Hasdemir Deniz , Kasper Dennis
Published: June 2021

Contribution of commensal microbiota to the host immune maturation gathers significant interests, however, their underlying mechanisms are remained to be investigated. We have focused on Bacteroides fragilis, one of the most ubiquitous residents of human gut. Establishing diverse, high‐sensitivity analysis methodologies, we have identified two commensal immunomodulatory factors, α‐galactosylceramides (BfαGCs) and a lipid anchor of polysaccharide A (PSA).

Previously, we have shown that glycosphingolipids from B. fragilis can regulate host natural killer T cell (NKT) number and administration of glycosphingolipids in early stage of life can protect animals from NKT‐mediated colitis. Among vastly diverse gut microbiota, ceramides, which includes glycosphingolipids, are almost exclusively produced by order Bacteroidales; LC‐MS/MS based metabolomic screening identified that α‐galactosylceramides are even more restricted in only a few commensal species, including B. fragilis. B.fragilis produces several classes of sphingolipids with diverse acyl chain length and branching. Among them, purified BfαGCs antagonize binding of prototypic ligands to CD1d and subsequent activation of NKTs in structure‐specific manner.

In addition to the previously reported immunomodulatory functions of PSA, the acid‐labile lipid anchor of PSA was critical to TLR2‐mediated IL‐10 production. MALDI‐TOF‐TOF analysis has identified that hydrophobic moiety resembles the structure of B. fragilis lipid A, with distinct difference in molecular composition, such as a degree of acylation. These results imply that PSA might bear a multiple functional moieties with distinct cellular signaling pathways.
Identification of Glutathione Adducts of the Myeloperoxidase Oxidation Product of Plasmalogen, α‐Chlorofatty Aldehyde
Authors: Duerr Mark , Ford David
Published: June 2021

α‐Chlorofatty aldehydes (α‐ClFALD) are produced by hypochlorous acid attack on plasmalogens during neutrophil activation. In this study the reactivity of the α‐chlorinated carbon of α‐ClFALD with the nucleophile, glutathione (GSH) was characterized, and the product of this reaction was examined in activated human neutrophils. Utilizing electrospray ionization mass spectroscopy, the reaction product of GSH and the α‐ClFALD, 2‐chlorohexadecanal, was characterized. The resulting conjugate, hexadecanal‐GSH retains an intact free aldehyde while the α‐chlorinated carbon is covalently bound to the sulfhydryl of GSH resulting in chlorine ejection from this adduct. Stable isotope labeled [d4]‐ hexadecanal‐GSH was synthesized. [d4]‐hexadecanal‐GSH was used to further confirm the structure, and was used to quantify natural α‐ClFALD conjugates of GSH using reversed phase liquid chromatography with detection by ESI‐MS and selective reaction monitoring. The 2‐chlorohexadecanal conjugate of GSH, hexadecanal‐GSH, is elevated in RAW 264.7 cells treated with physiologically relevant concentrations of exogenous 2‐chlorohexadecanal. Furthermore, phorbol myristate acetate (PMA) treated primary human neutrophils have elevated levels of hexadecanal‐GSH, octadecanal‐GSH, 2‐chlorohexadecanal and 2‐chlorooctadecanal levels, which are reduced in PMA treated neutrophils pretreated with aminotriazole. Together, these studies demonstrate for the first time that chlorinated fatty aldehydes are covalently modified by glutathione forming novel peptidoaldehydes, which may play an important role in inflammatory signaling.
Determination of Height Using Femur Length in Adult Population of Oguta Local Government Area of Imo State Nigeria
Authors: Obialor Ambrose , Ihentuge Churchill , Akpuaka Frank
Published: June 2021

The use of body parts in the estimation of stature is a vital tool in forensic anthropology. This research was carried out to determine the height of adult population of Oguta inhabitants using the femur length. The femur length of 200 adults (male 123, female 77) aged 22 and 70 was taken using a non stretchable tape to measure the distance between the greater trochanter as palpated and lateral condyle of the femur. Height was taken from the subject by a calibrated meter rule. The mean of femur length and height was 41.18cm ± 0.03 and 164.47cm ± 0.65 respectively for the sample population in the male, the female length and height was 41.79cm ± 0.16 and 167.50cm ± 0.78 respectively, while in the female population the femur length was 40.82cm ± 0.26 and the height in 161.90cm ± 0.95. The pair of variables was found to be significant in each case at the level of p> 0.05. The value of the correlation factor is 0.9866. In this study the femur/length ratio for the sample population is 25. The mean femur/height ratio for male is 25.11 and that of female is 25.25. These values vary from reports from other parts of the world. This variation could be from the nutritional difference, occupation and the geographical location. The regression coefficient for the sample population is 0.9867. In the male population the value of r is 0.9927 while in the female population it is 0.9126. The values of r in this study showed that there is high correlation between the femur length and height of an individual. This study revealed that the height of an individual is four times the femur length of the individual, since the femur length is proportional to the height, therefore the taller the individual the longer femur length, while the shorter the individual the shorter the femur length.
Translational and Morphological Effects of Signalling Alcohols on C. albicans
Authors: Egbe Nkechi , Ashe Mark
Published: June 2021

Candida albicans is a polymorphic yeast that can cause life threatening systemic infections in immunocompromised individuals. One key attribute of C. albicans that enhances pathogenicity is the capacity to switch morphology from vegetative to filamentous modes in response to specific environmental conditions. Stressful changes in such cellular conditions commonly cause a rapid inhibition of global protein synthesis leading to altered programmes of gene expression.

Fusel alcohols induce hyphal/pseudohyphal growth, while farnesol, a quorum sensing alcohol, prevents this distinct form of growth. A variety of cell biological and genetic assays suggest that fusel alcohols and ethanol inhibit protein synthesis in C. albicans by targeting the translation initiation factor eIF2B, whereas farnesol inhibits protein synthesis in a manner that does not impact upon eIF2B activity. Rather biochemical and mass spectrometric analysis suggest farnesol affects the interaction of the mRNA with the small ribosomal subunit during translation initiation. Further analysis of the effect of farnesol on C. albicans transcript levels and ribosome association by next generation sequencing provide insight into genes that are differentially expressed following farnesol treatment. While genes involved in morphological differentiation, biofilm formation, cytokinesis and cell cycle were generally repressed, those involved in protein synthesis, oxidative stress and cellular respiration were upregulated. Intriguingly, the regulation of these functional categories of genes occurs in a co‐ordinated manner; at either the transcript level or at the level of ribosome association, but rarely at both transcriptional and post‐transcriptional levels for the same gene.
ApoA‐IV Regulates Cytokine Responsiveness in LPS‐challenged Macrophage Cells
Authors: Li Xiaoming , Li Zongfang , Yan Jianqun , Tso Patrick
Published: June 2021

ApoA‐IV has an anti‐inflammatory role but it is unknown whether apoA‐IV affects inflammation cytokines induced by LPS in macrophage cells.We determined if apoA‐IV influences the secretion of cytokines stimulated by LPS in the cells.

The macrophage RAW264.7 cells were pre‐treated with apoA‐IV (20µg/ml) for 1h, followed with LPS (500ng/ml) for 20 h. The culture medium was collected and RayBio® Mouse Inflammation Antibody Array G Series I was used to measure the cytokines.

Both apoA‐IV and LPS or the two of them have no effect on the cytokines: POS, NEG, Lymphotactin, MCSF, MIP‐1α, MIP‐1γ, IL‐13, IL‐1β, BLC, CD30L, Eotaxin, IFN‐γ, and MIG, in 42 cytokines detected. ApoA‐IV had the trend to reduce Fas ligand, Fractalkine and Eotaxin‐2. Both apoA‐IV and LPS increased the cytokines GCSF, IL‐2, IL‐3, IL‐4, IL‐6, IL‐9, IL‐12P40/P70, GCSF, TIMP‐1, LIX, SDP‐1, TCA‐3, TECK, TIMP‐2, sTNF‐RI, sTNF‐RII, GM‐CSF, RANTES, I‐TAC, KC, MCP1,TNF‐α, TECK, IL‐17 and Leptin. Notably, TNF‐α and TECK induced by LPS were significantly inhibited by apoA‐IV. Interestingly,apoA‐IV enhanced IL‐10 and IL‐1α by itself and that induced by LPS.

In conclusions, both LPS and apoA‐IV can induce many inflammation cytokines from macrophage cells, but the patterns are different. ApoA‐IV negatively or positively regulates a few cytokines induced by LPS. The mechanism involved remains to be elucidated. Supported by Program for Changjiang Scholars and Innovative Research Team (PCSIRT:1171).
Ras Isoforms Conformational Clustering and Community Networks Studies: Simulating Ras with Accelerated Molecular Dynamics
Authors: Guterres Hugo , Ma Buyong , Nussinov Ruth , Mattos Carla
Published: June 2021

Ras is a small GTPase that acts as a molecular switch and is involved in multiple cellular signaling pathways. The catalytic domains of the three major isoforms, H‐, K‐, and N‐Ras share an identical effector lobe, and 90% sequence identity in their allosteric lobe. Ras isoforms have been shown experimentally to exhibit functional specificity yet they display very similar three‐dimensional structures. In this study, we used accelerated Molecular Dynamics (aMD) to characterize the isoform‐dependent conformations of Ras. The aMD method enhances conformational sampling by reducing the energetic barriers between states of a system. Our results demonstrate a unique cluster of conformations between Ras isoforms. Additionally, we used community network analysis to probe the communication network between the effector and allosteric lobes across the three isoforms. We found that H‐ and K‐Ras share similar sets of community networks, whereas N‐Ras exhibits a unique set of community networks. These data on conformational clustering and community networks in Ras isoforms advance our understanding of the structural biology behind their functional specificity.
Defining Starch Binding by Glucan Phosphatases
Authors: Auger Kyle , Raththagala Madushi , Wilkens Casper , Svensson Birte , Gentry Matthew
Published: June 2021

Starch is a vital energy molecule in plants that has a wide variety of uses in industry, such as feedstock for biomaterial processing and biofuel production. Plants employ a three enzyme cyclic process utilizing kinases, amylases, and phosphatases to degrade starch in a diurnal manner. Starch is comprised of the branched glucan amylopectin and the more linear glucan amylose. Our lab has determined the first structures of these glucan phosphatases and we have defined their enzymatic action. Despite this progress, we lacked a means to quickly and efficiently quantify starch binding to glucan phosphatases. The main objective of this study was to quantify the binding affinity of different enzymes that are involved in this cyclic process. We established a protocol to quickly, reproducibly, and quantitatively measure the binding of the enzymes to glucans utilizing Affinity Gel Electrophoresis (AGE) and Surface Plasmon Resonance (SPR). The results show that the glucan phosphatases possess differing abilities to bind to different glucan substrates. One glucan phosphatase possesses a 50 fold higher affinity for amylopectin than the other, while it only possessed a 3 fold higher affinity for amylose. Both glucan phosphatases showed similar affinities for the short oligosaccharide β‐cyclodextrin. We performed structure‐guided mutagenesis to define the mechanism of these differences. We found that the carbohydrate binding module (CBM) domain provided a stronger binding affinity compared to surface binding sites (SBSs).
Nuclear Export of Spliceosome‐discarded Intermediates
Authors: Zeng Yi , Staley Jonathan
Published: June 2021

The precise removal of introns is essential for cell survival. To promote fidelity in nuclear pre‐mRNA splicing, the spliceosome rejects and discards suboptimal splicing substrates that have engaged the spliceosome. Although intron‐containing transcripts can be retained and degraded in the nucleus, various spliceosome‐discarded intermediates, such as mutated lariat intermediates, can be degraded in the cytoplasm. However, it is unknown why or how these suboptimal intermediates are exported for turnover.

Here, we used RNA‐FISH to provide direct evidence that mutated lariat intermediates are exported into the cytoplasm and disruption of DEAH‐box ATPase Prp43p‐mediated discard of suboptimal intermediates blocks the export of mutated lariat intermediates. Further, we demonstrated that nucleoporins (Nup2p and Nup60p) and the general mRNA export receptor, Mex67p, play an essential role in the export of mutated lariat intermediates. Surprisingly, we found that Mlp1p, the nuclear retention factor, is important for the export of mutated lariat intermediates. Together, our findings establish the importance of the mRNA export pathway in transporting discarded intermediates for degradation.
User Variance in Defining the Trabecular‐Cortical Interface Using MicroCT Images
Authors: Bryce Amy , Terhune Claire , Sylvester Adam
Published: June 2021

Micro‐computed tomography (µCT) imaging is an effective tool for quantifying bone microarchitecture. µCT images allow trabecular and cortical bone to be separated, a task critical to understanding bone mechanics. Automated algorithms are often used to facilitate this process, but are typically evaluated by comparing algorithm results to those of a single person. Such analyses do not take into account the possibility of variability among observers in defining this boundary.

This study quantified the amount of inter‐observer variation in defining the cortical‐trabecular bone boundary. Ten µCT images of human femora and primate mandibles were sent to 19 researchers with interests in bone biology. Participants were asked to trace the cortical‐trabecular boundary on each image as well as to complete a questionnaire about education, training, and specific areas of interest in bone biology. For each image, boundary lines were converted to semilandmarks for use in geometric morphometric analyses. Shape variation was summarized using principle components analysis and variation among users was quantified.

All of the µCT images exhibited variance in where the participants defined the cortical/trabecular interface. Often the variation was small (<10 voxels), but in some images there was considerable variation in boundary location. Analyses did not reveal significant differences (p>0.05) in boundary contours based on different levels of education or areas of expertise. The results do, however, indicate variability among bone biologists in defining the trabecular‐cortical boundary and question the validity of algorithms evaluated against a single user.
Intermittent Fasting of High‐Fat Diet Increases Hypothalamic Norepinephrine and Improves Metabolic Parameters in Obese Mice
Authors: Gotthardt Juliet , Verpeut Jessica , Yeomans Bryn , Roepke Troy , Bello Nicholas
Published: June 2021

Alternate day, intermittent fasting (IMF) can promote weight loss in obese individuals. We investigated the metabolic and neural mechanisms of IMF in diet‐induced obese mice. Twenty‐four C57 male mice at PND 49 were fed a high‐fat diet (HFD; 45% fat, 20% protein, 35% CHO) ad libitum for 8 wks. Following this, for 4 wks, mice (n = 8/group) were maintained on ad libitum HFD, received IMF of HFD (IMF‐HFD), or were switched to ad libitum low‐fat diet (LFD;10% fat, 20% protein, 70% CHO). Body weight and cumulative calorie intake were not different between the IMF‐HFD and LFD. Indeed, both groups had reduced cumulative caloric intake (~13% for both; p < 0.01) and reduced body weight (~10% for both; p < 0.05) compared with the HFD. Metabolic parameters, however, were different between the IMF‐HFD and LFD groups. Insulin tolerance was improved in IMF‐HFD compared with the HFD group (‐ 20% glucose AUC; p < 0.05), but impaired compared with LFD (+ 22% glucose AUC; p<0.05). Following a fast (8 h), ketone bodies were elevated in IMF‐HFD compared with HFD (p < 0.05) and LFD (p = 0.05). Indirect calorimetry revealed lower RER in IMF‐HFD compared with HFD (p<0.001), but higher than LFD (p<0.05). Hypothalamic tissue homogenates indicated higher levels of norepinephrine in the paraventricular nucleus region of IMF‐HFD compared with HFD (+40.6%, p<0.05) and LFD (+31%, p = 0.05) animals. Thus, while IMF‐HFD reduces body weight and calorie intake similar to a LFD, weight loss by IMF produces distinct metabolic and neural alterations.
Establishing Regression Parameters to Simplify Determination of Carbohydrate Intolerance
Authors: Levers K , Rasmussen C , Greenwood M , Earnest C , Kreider R
Published: June 2021

Determining carbohydrate intolerance (CI) and insulin sensitivity is vital when selecting the proper diet to maximize results and improve health. This study examined answers to a CI questionnaire (CIQ) and simple health measures to develop a quick method to determine accurate prediction, non‐prediction models of CI vs. traditional OGTT outcome measures. 108 women (31.6±13 yrs, 34.7±7% body fat, 25.3±4 kg/m2) donated fasting blood samples, completed a CI inventory, had DEXA body composition and health measures determined, and underwent a 75g, 2‐hr OGTT. Considering previous Pearson correlations, numerous linear regression models were run to build the best prediction model of CI questions plus health measures for OGTT glucose at 120 minutes (G120) and glucose AUC (GAUC). Results revealed that 2 models demonstrated positive associations for total yes responses to 7 CI questions + 3 health measures: experience headaches, fatigue/exhaustion, crave starchy/sugary foods, drowsiness, meal time hunger, eat 3 meals/day, maintain constant extra body wt, age, body wt (kg), and BMI. G120 = 2.649(CI total yes)+0.019(age)‐1.399(wt)+7.584(BMI), r2=0.949. GAUC = 7.781(CI total yes)+0.248(age)‐2.16(wt)+14.253(BMI), r2=0.955. The basic 7 question CI inventory alone (total yes responses) also showed a strong association with G120 (r2=0.866) and GAUC (r2=0.880). These findings show promise for simple CI results without an OGTT.
Maternal low protein diet leads to dysregulation of placental iNKT cells and M1/M2 macrophage ratio, body weight loss in male, neonate Sprague‐Dawley rats and increased UCP‐1 mediated thermogenesis
Authors: Vomhof‐DeKrey Emilie , Roemmich James , Claycombe Kate
Published: June 2021

Placental immune cells provide cytokines and growth factors that are necessary for placenta development and function. Invariant natural killer T (iNKT) cells are innate cells specific for glycolipid antigens presented by the CD1d molecule and secrete Th1 cytokines in the placenta, suggesting an immunoregulatory role. Self‐lipid antigens at the maternal‐fetal interface may activate iNKT cells, leading to placental inflammation and dysfunction, which can increase brown adipose tissue (BAT) thermogenesis and reduce fetal growth. Maternal low protein (LP) diets reduce offspring birth weight (BW) perhaps via a BAT thermogenesis‐mediated effect. However, whether maternal LP nutrition alters placental immune cell populations and offspring thermogenesis in not yet known. Obese‐prone Sprague‐Dawley dams were fed 8% LP or 20% normal protein (NP) diets for 3 wks prior to breeding and through pregnancy. BW of neonatal LP males was lower than NP males. Vα14 mRNA marker for iNKT cells was 11‐fold higher in LP placenta, while there was also an increase in proinflammatory M1/M2 macrophage mRNA marker ratio. UCP‐1 gene and protein expression and transcription factors PRDM16 and PPARα in BAT were 2‐ to 6‐fold greater in LP than NP neonatal males. FNDC5, an irisin precursor and thermogenesis activator, was 2‐fold greater in neonatal LP BAT than NP males. These results suggest that prenatal protein restriction increases proinflammatory iNKT and M1 macrophages in the placenta, leading to placental dysfunction, increased offspring BAT thermogenesis, and low BW.
Dietary IGF‐activated Plasma Protein Alleviates Intestine Dystrophy by Induction of Tight Junction Protein in Early Weaned Rats
Authors: Kwak Minjin , Kim Jonggun , Kim Hyung‐Kwan , Ha Dong‐Jin , Whang Kwang‐Youn
Published: June 2021

Weaning is a stressful condition for mammals and negatively affects growth performance by dystrophy of intestinal villi and crypts. Dietary plasma protein (PP) has shown the increased growth performance in early‐weaned animals and it seems to be more than nutrient content itself. Among growth factors, dietary insulin‐like growth factor (IGF) known to enhance growth performance, but bound form of IGF might reduce growth promoting effects of IGF. The effects of IGF‐activated plasma protein (aIGFPP, by de‐binding with proteins) on growth performance and gut development were studied in early‐weaned rats.

Twenty‐one rats (weaned at 14‐day) were allotted into three treatments (NC, NIH‐31 diet without animal proteins; PC, NIH‐31 diet with 50 g/kg PP; and IGF, NIH‐31 diet with 3.4 g/kg aIGFPP). After 6 days, jejunum samples were collected and gene expression of inflammatory cytokines (IL‐6, TNF‐α, IL‐1β, IL‐4, IL‐12β, and IL‐10) and tight junction proteins (β‐catenin and ZO‐1) were determined by qRT‐PCR. Body weights were measured daily. And the villus heights and crypt depths were also measured in jejunum.

Body weight was numerically higher in rats of PC and IGF groups than rats of NC. There was no differences in TNF‐α, IL‐1β, IL‐4, IL‐12β, and IL‐10 expression, but IL‐6 was higher in IGF group than other groups (P<0.05). Tight junction protein, ZO‐1 and β‐catenin, expression was higher in IGF group than other groups at P<0.10 and P<0.05, respectively. Jejunal villus heights of rats in IGF group were higher than rats in other groups (P<0.05) but no difference was found in crypt depth. This data indicates aIGFPP could increase gut growth and might affect growth performance in early‐weaned rats.
Oxidative Stress, Nutrient Intake, and Red Blood Cell Membrane Fatty Acid Profiles of Women with and without Fertility Issues
Authors: Litwin Nicole , Assad Norman , Clark W. Andrew , Mohseni Ray , Zheng Shimin , Ferrell Tasha , Santos‐Prowse Robert
Published: June 2021

This study examined nutritional status and fecundity of women of reproductive age, as infertility affects 11% of this population. Participants filled out anthropometric and food frequency questionnaires (FFQ) and had 5 mL sample of whole blood collected for measures of oxidative stress (OS) and fatty acid (FA) profiles of the red blood cell (RBC) membrane. Participants designated to the case group (n=30) were women experiencing fertility issues with an anti‐Mullerian hormone level of 蠄1 ng/ml and those with normal ovulation (n=30) were designated to the control group. Our hypothesis was that the case group would have greater OS, consume more saturated fat and have lower omega‐3 FA levels and higher omega‐6 FA levels in RBC membranes. Results showed that case participants had significantly higher body mass index (BMI) when graduating high school (22.9 vs. 20.86; p=0.027) compared to study enrollment (28.47 vs. 23.14;p=0.005) and higher weight gain since high school (32. 6 lbs vs. 14.2 lbs; p=0.006). FFQ results indicated case participants consumed significantly higher amounts of total fat (100.14 vs. 86.7 g/d; p=0.005), saturated fat (26.53 vs. 21.93 g/d; p=0.011), unsaturated fat (73.6 vs. 64.7 g/d; p=0.003), and had a higher percentage of total dietary fat (36.2% vs. 32.8%; p=0.005) in their diets. No significant differences were noted in oxidative stress levels of plasma or RBC membrane FA types and ratios between the two groups.
Healthy Eating Index Scores are Associated with Inflammatory Markers in Breast Cancer Survivors
Authors: Arnold Kristen , Andridge Rebecca , Logan Amanda , Yee Lisa , Lustberg Maryam , Orchard Tonya
Published: June 2021

Lower inflammatory markers are associated with improved survival in women diagnosed with early stage breast cancer. Our objective was to determine if diet quality was associated with inflammation in breast cancer survivors. Food frequency questionnaires from a randomized placebo‐controlled pilot study comparing omega‐3 fatty acid supplementation versus placebo in 44 postmenopausal women with breast cancer beginning aromatase inhibitor (AI) treatment were analyzed and scored for diet quality using the Healthy Eating Index 2010 (HEI). Pearson correlation coefficients (r) were used to evaluate the relationship between baseline HEI scores and inflammatory cytokines, interleukin‐6 (IL‐6), IL‐17, and tumor necrosis factor alpha receptor 2 (TNFR2), at baseline (0), 12 and 24 weeks after beginning AIs. Complete data were available for 34 women at week 24. HEI scores did not differ by randomization or chemotherapy status at baseline (P=0.23). Higher baseline HEI score (healthier diet) was associated with significantly lower IL‐6 (Week 0 and 24: r = ‐0.42 and ‐0.51 respectively; P<0.002) and TNFR‐2 (Week 0 and 24: r = ‐0.35 and ‐0.41 respectively; P<0.02) in analysis of all participants. In women who had prior chemotherapy (n=16), stronger inverse relationships between HEI, IL‐6 (Week 0 and 24: r = ‐0.62 and ‐0.84; P<0.01) and TNFR‐2 (Week 0 and 24: r = ‐0.55 and ‐0.67; P<0.03) were observed. We found no significant relationship between HEI and IL‐17. These data suggest that healthier diet quality is associated with some markers of decreased inflammation in breast cancer survivors beginning AI therapy.
Isoproterenol Induces Beiging and Thermogenesis in Mature 3T3‐L1 Adipocytes
Authors: Miller Colette , Yang Jeong‐Yeh , England Emily , Yin Amelia , Gajjar Viral , Rayalam Srujana
Published: June 2021

Prolonged exposure to cold temperature has shown to induce beiging of white adipose tissue in vivo. However, an in vitro model to study the mechanisms underlying the transdifferentiation ability of white to beige adipocytes is lacking. To develop such a model, 3T3‐L1 cells, the most commonly used murine adipocyte cell line, were induced to differentiate using insulin, T3, IMBX, and dexamethasone. Mature adipocytes were then treated with the beta‐agonist isoproterenol (10 or 100 µM) for up to 48 hours. Isoproterenol increased the transcription of UCP1 and beige markers CD137, CITED1, and HOXC8. The induction of UCP1 and CD137 were confirmed using immunocytochemistry. Lastly, Oxygen Consumption Rate (OCR) was measured using the Seahorse Bioanalyzer. High‐dose isoproterenol increased OCR at multiple time points, leading to an increased total OCR when area under the curve was analyzed (p=0.06). Our research demonstrates that mature 3T3‐L1 adipocytes upon stimulation can adopt a beige genotype and increase thermogenesis. Furthermore, these cells can be an efficacious model to understand the process of white to beige adipocyte transdifferentiation and to screen pharmaceutical/nutraceutical agents for beiging properties. This research was supported by the Georgia Research Alliance Eminent Scholar Endowment, Atlanta Chapter of Achievement Rewards for College Scientists, and with CSO Intramural Funds from the Philadelphia College of Osteopathic Medicine ‐ School of Pharmacy.
Effects of Caffeine on Peroxynitrite‐Mediated DNA Strand Breakage
Authors: Babu Pon Velayutham Anandh , McCormick John , Jia Zhenquan
Published: June 2021

Caffeine is a widely consumed beverage in much of the world. Recent epidemiological studies have demonstrated that coffee consumption is inversely associated with Parkinson's disease. However, the mechanism for this protection is unknown. Oxidative stress has been suggested to play an important role in neuronal degeneration. Peroxynitrite is a strong oxidizing agent that is generated from the bi‐radical reaction of nitric oxide and superoxide at a diffusion‐limited rate leading to neurotoxicity. Peroxynitrite has been found in various cell types including both neurons and glial cells in the brain of Parkinsonian patients. DNA strand breakage induced by peroxynitrite‐mediated oxidative stress has been suggested to be a cause of neurodegenerative disorders. Using phi X‐174 plasmid DNA as an in vitro system, this study was undertaken to investigate the effects of caffeine in peroxynitrite‐induced DNA strand breaks, a critical event leading to peroxynitrite‐elicited cytotoxicity. Incubation of phi X‐174 plasmid DNA with the 3‐morpholinosydnonimine (SIN‐1), a peroxynitrite generator, led to the formation of DNA break in a concentration‐ and time‐dependent fashion. The presence of caffeine at concentrations ranging from 3.2 µM to 3.20mM was found to significantly inhibit SIN‐1‐induced DNA strand breaks in a concentration‐dependent manner. DNA was also incubated with other well‐known antioxidants to compare the inhibition of peroxynitrite‐induced damage by caffeine. The protective effects of caffeine on peroxynitrite‐mediated oxidative DNA damage may shade some light on the mechanism(s) of its neuroprotective activities observed in human clinical trials.
Super‐resolution Imaging Shows Spatial Regulation of p130Cas Phosporylation within Focal Adhesions
Authors: Elmokadem Ahmed , Moraru Ion , Yu Ji
Published: June 2021

Cell motility is critical for physiological processes such as tissue development and remodeling as well as pathologies such as cancer metastasis. Cell motility is influenced by its surrounding matrix through biochemical and mechanical stimuli. p130Cas is a signaling molecule that has been proposed to be a mechanosensor, which responds to mechanical stimuli and mediates cell motility during the early stage of lamellipodial protrusions. p130Cas localizes to focal adhesions (FAs) and binds with its effectors via tyrosine phosphorylation of its substrate domain. Previous studies have suggested that mechanical stimuli increase p130Cas tyrosine phosphorylation and these phosphotyrosines recruit SH2‐domain containing effectors like Crk, that initiate a signaling cascade activating the small Rho‐GTPase Rac1 leading to lamellipodial protrusions.

Due to spatial heterogeneity of stresses within FAs, we hypothesized that p130Cas phosphorylation is not uniform across a single FA. Spatial differences across such small structures as FAs cannot be resolved using biochemical assays or standard microscopy. We used a super resolution imaging technique, PALM (photo‐activated localization microscopy) to study the differences in p130Cas phosphorylation within individual FAs using SH2 domains of Crk as probes. Our results show punctate localization of SH2 probes at single FAs compared to uniform distribution of p130Cas.

Our data suggest that p130Cas activity is spatially regulated. In the future we will look at correlations between mechanical stress within FAs and p130Cas activity. Our results will help elucidate the mechanism of mechanical regulation of p130Cas phosphorylation, and may help understand how cells sense and respond to external mechanical stimuli.
The preference and intake frequency of processed food among university students by residence type
Authors: Kim Se‐Yune , Choi Mi‐Kyeong , Kim Su‐Jin
Published: June 2021

The purpose of this study is to assess the intake status of processed food in university students by residence type. This cross‐sectional study was conducted by surveying 476 university students in Korea. The preference for and intake frequency of processed foods were examined through questionnaires, and analyses were conducted by comparing the results by residence type (home with parents, n=120; self‐boarding, n=170; lodging, n=68; and boarding with meals, n=118). Approximately 42% of the self‐boarding group responded that processed food was easy to cook, and the highest priority factor they checked when purchasing processed food was the price. Among processed foods, subjects had the highest preference for noodles, and the home with parents group and the lodging group showed the highest preference for milk and dairy products. The self‐boarding group and the boarding with meals group reported the highest preferences for biscuits, retort pouch food, convenience food, and canned food. The self‐boarding group showed a high frequency of processed food intake in comparison with the other residence groups, which may cause health problems because these individuals are taking in unbalanced nutrients. These results suggest that nutritional education related to processed food is needed for university students living in various residence types.
Dietary Supplement Use Among the Undergraduate College Population
Authors: Freeman Hanna , Kam Lynn , Harris Stephanie
Published: June 2021

Over the past decade, the prevalence of dietary supplement use in the United States has increased among the general adult population.

Purpose

To determine the relationship among dietary supplement use, academic major, health information source, and motivation for use in the undergraduate college population.

Methods

Undergraduates at Case Western Reserve University were invited to participate in this study. A survey was administered through REDcap, a secure web‐based database, to interested participants. The survey included questions on (i) gender, (ii) intended academic major, (iii) dietary supplement use, (iv) rational for use, and (v) sources of information on dietary supplements.

Results

In total, 238 undergraduates completed the survey with 71% reporting dietary supplement use. Nutrition majors and pre‐health majors were as likely to use dietary supplements as other academic majors (odds ratio=1.11, 95% CI for odd ratio= 0.63‐ 1.98). Main motivations for use were to improve or maintain overall health (88%), to boost immunity (50%), and for a vitamin/mineral deficiency (32%). The top three sources used to obtain information on dietary supplements were the Internet (57%), family or friends (49%), and physicians (38%).

Conclusions

Dietary supplement use is high in the undergraduate student population. Most students reported using unreliable resources to obtain information on dietary supplements, and thus may be potentially making poor health choices. In addition, many health majors are planning for careers in the medical profession. Therefore, critical assessment of reliable health information should be reinforced in their academic curriculums, so that they can provide evidence‐based advice to their future patients.
Calcein's Quenching In Vitro Method for Assessing Dietary Iron Bioavailability
Authors: Ye Shumao , Armah Seth , Xu Qi , Reddy Manju
Published: June 2021

Low iron bioavailability (IBA) is a major contributor of iron deficiency anemia, a most prevalent nutritional deficiency around the world. In vitro IBA is commonly measured using simulated gastrointestinal digestion, caco‐2 cell uptake and followed by ferritin response measurement. This method is cumbersome since it takes more than 24 hours and needs an expensive kit to measure ferritin concentrations. Hence, our objective was to develop a high throughput and cost effective method to assess IBA in large number of samples in a short period of time. In this new method, the quenching of calcein fluorescence in a caco‐2 cell model by food iron following in vitro digestion is used as an index of IBA. The meals we used were similar to the ones that were used in human studies. IBA assessed in this method was compared with published human absorption data and measured ferritin responses. Similar to human absorption data, IBA was enhanced by ascorbic acid, and was reduced by bran, phytate, and tea. A high correlation was found between published human absorption data and the IBA we tested (r=0.90; p=0.04). Our data was also highly correlated with the data obtained using traditional ferritin method (r=0.92; p<0.01). In conclusion, our method does not only reliably predict human iron absorption, but also is a more cost effective and faster one compared to the widely used ferritin method. However, future studies are needed for more comparisons with human studies.
Cell Membrane Disruption and DNA Binding of Staphylococcus Aureus Induced by Antibacterial Peptide F1 from Tibetan Kefir
Authors: Miao Jianyin , Fan Weihao , Zhou Jianliang , Liu Guo , Chen Feilong , Chen Yunjiao , Gao Xiangyang , Dixon William , Xiao Hang , Cao Yong
Published: June 2021

The purpose of this work was to investigate antimicrobial mechanisms of a unique peptide F1 (found in Tibetan Kefir) against Staphylococcus aureus. The cell membrane permeability assay and potassium ion release assay were employed to examine the integrity of the bacterial membrane after the treatment with antimicrobial peptide F1. The transmission electron microscopy was used to analyze the effect of antimicrobial peptide F1 on the cell morphology of Staphylococcus aureus. The interaction between antimicrobial peptide F1 and genomic DNA of Staphylococcus aureus was investigated by DNA gel retardation assay and atomic force microscopy imaging analysis. The results showed that treatment with the antimicrobial peptide F1 led to the permeabilization of the cell membrane of Staphylococcus aureus. Moreover, the antimicrobial peptide F1 can bind to the DNA and inhibit related cellular functions. Having multiple targets means that antimicrobial peptide F1 may be more effective in killing bacterial cells, and bacteria have additional hurdles to overcome to build resistance. These results indicate that antimicrobial peptide F1 is a potential candidate as an effective biological preservative in the agricultural and food industry.
Effects of Calcium and Vitamin D Supplementation on Markers of Colonic Permeability
Authors: Fedirko Veronika , Bostick Roberd , Gewirtz Andrew , Tran Hao Quang , Barry Elizabeth , Baron John
Published: June 2021

Impaired gut barrier function contributes to inflammatory bowel diseases and colorectal carcinogenesis. Recent evidence from experimental models indicates that calcium and vitamin D may beneficially modulate colonic permeability; however, to date, there are no reported human studies that investigated this hypothesis. To address this, we tested the effects of calcium (1.2 g/d) and/or vitamin D3 (1,000 IU/d) supplementation for 3 or 5 years on circulating biomarkers of colonic hyperpermeability (anti‐flagellin and anti‐lipopolysaccharide immunoglobulins [Igs], IgA and IgG) in 175 participants with previous colorectal adenoma from a recently completed, multi‐center, randomized, double‐blind, placebo‐controlled, modified 2 x 2 factorial chemoprevention clinical trial (NCT00153816). Igs were measured in plasma via ELISA. The relative change in a combined permeability score (CPS, defined as the sum of the optical densities of all biomarkers) from baseline to follow‐up was ‐2%, 1%, and 4% in the calcium, calcium plus vitamin D, and vitamin D groups vs. placebo, respectively (all P‐values > 0.5). At baseline, the CPS was higher among men (P = 0.02) and the severely obese (BMI >35 kg/m2; P = 0.002). Our preliminary results indicate no effect of calcium and vitamin D treatment on markers of colonic permeability, and suggest that men and those who are obese may have greater gut permeability.
Relationship between expended energy and stored energy as lean and adipose tissues in growing rats
Authors: Tai Yuki , Fujii Takako , Maeda Megumi , Okamura Koji
Published: June 2021

Since the energy contents differ between lean tissue (1,050 kcal/kg) and adipose tissue (7,600 kcal/kg) and lean tissue is considered to expend a greater amount of energy for the synthesis of a greater amount of protein, the energy expended for growth varies theoretically depending on the amount and the ratio of the increases in these two tissues. Five growing rats (4 weeks old, body weight (BW) 102 g (SD 9)) were fed a standard diet ad‐libitum and the BW, total energy consumption (TEC) and expenditure (TEE) were measured for 14 days. The energy stored in the body (EST) was calculated by subtracting TEE from TEC. TEC, TEE and EST were 1,081 kcal (SD 92), 622 kcal (SD 32), 458 kcal (SD 80), respectively. Forty‐two percent of the energy consumed was stored in the body, while 58% of it was expended. As the BW increased by 135 g (SD 10), the energy stored for 1 kg of the body was calculated to be 3,370 kcal (SD 406). Based on the energy contents in the lean tissue and the adipose tissue and the EST, the estimated increases in the lean tissue and the adipose tissue appeared to be 87 g and 48 g, respectively, and the ratio of the increases was 64:36 in growing rats.
The Role of Glucose Metabolism in Cardiac Remodeling Induced by Tobacco Smoke Exposure
Authors: Azevedo Paula , Ferreira Vanessa , Santos Priscila , Gonçalves Andrea , Okoshi Marina , Fernandes Ana Angelica , Roscani Meliza , Polegato Bertha , Paiva Sergio , Zornoff Leonardo , Minicucci Marcos
Published: June 2021

Therefore The aim of this study is to investigate the cardiac glucose metabolism pathways in cardiac remodeling induced by TS.

Methods 20 animals were allocated into 2 groups: group of animals exposed to tobacco smoke (TS) and group animals not exposed to cigarette smoke (C). After 2 months the animals were subjected to: a) echocardiographic study; e) spectrophotometric evaluation of energy metabolism enzymes and f) Western blot for Akt and AMPK.The TS group showed larger left atrium size and lower ejection fraction than those of group. The animals showed the same body weight (p = 0.3), heart rate ( p = 0.4) and mean arterial pressure. As regards the analysis of energy metabolism enzymes, phosphofructokinase (PFK) (C 138 ± 24, TS=198 ± 31, p = 0.003), LDH (C =121 ± 31, TS= 167 ± 27, p = 0.017) and pyruvate dehydrogenase (PDH) (C=208 ± 18, TS= 211 ± 27, p=0,83) were greater in the TS group and citrate synthase (C =39,0 ±2.32, 25.53 ± TS=5.36, p <0.001), complex I (C =5.03 ± 0.48, TS=2.75 ± 0.62, p <0.001), complex II (C=5.32 ± 0.93; TS= 2.48 ± 0.78, p <0.001) , ATP synthase (C=57.91 ± 7.24, TS= 21.18 ± 4.84 p <0.001) were lower in the TS group. Akt/Aktp was similar between the groups. AMPK was reduced in CS group (p<0,001).Conclusion: During early cardiac remodeling, there is an increase in glucose metabolism. In this study LDH and PFK , that represents glycolytic pathways were increased. However, the citric acid cicle and mitochondrial respiratory chain may be compromised. One of the key modulators of these features may be AMPK.
Phosphorus supplementation recovers the blunted diet induced thermogenesis in overweight and obese subjects
Authors: Bassil Maya , Obeid Omar
Published: June 2021

Diet induced thermogenesis (DIT) of obese subjects was reported to be blunted or lower than that of lean subjects (Ravussin & Swinburn, 1996). We have previously found that phosphorus (P) supplementation increases DIT of glucose solution in healthy lean subjects (Bassil et al, 2013) probably due to an increase in postprandial ATP production. A study was conducted to investigate whether P supplementation is able to stimulate DIT of overweight and obese subjects. DIT of glucose solution was measured with or without P in 12 overweight and obese subjects (4M, 8F; Age=22.4±1.2y; BMI= 30.3 ±0.9kg/m2) in a double‐blind cross over design with 1 week washout period. After a 10‐h overnight fast, subjects drank a standardized 296mL solution containing 75g glucose, with P (500mg) or placebo pills. Oxygen consumption (VO2) was measured every 30 min for 3 hours using an indirect calorimeter to derive energy expenditure. Baseline fasting resting metabolic rate (RMR) and VO2 did not differ between the 2 experiments (p=0.4). In line with the literature, DIT of overweight and obese subjects was blunted with placebo, whereby VO2 did not change from baseline after the consumption of glucose solution. Conversely, P supplementation was associated with a significant increase (p<0.05) in VO2 during the first 90 min (320±17 mL/min) compared with baseline (297±10 mL/min) which was equivalent to a 10% increase in DIT from baseline. In conclusion, P supplementation recovers the blunted DIT in overweight and obese subjects. These results highlight the important role of P in energy balance in obesity. Further studies are warranted to elucidate the long term effects of P supplementation and to explore potential mechanisms of action.
Prevalence of Genetic Hemoglobin Variants and Relationships with Anemia and Micronutrient Status among Children in Urban Cameroon
Authors: Engle‐Stone Reina , Nankap Martin , Ndjebayi Alex , Brown Kenneth , Green Ralph
Published: June 2021

Information on the causes of anemia is needed to develop effective anemia‐control programs. Our objective was to measure the prevalence of genetic hemoglobin variants in a representative sample of children in urban Cameroon, and examine the relationship between these variants and both anemia and micronutrient status indicators. In a 2012 survey of 306 clinically asymptomatic children aged 12‐59 months in Yaoundé and Douala, Cameroon, we assessed malaria infection, inflammation (C‐reactive protein and α1‐acid glycoprotein), hemoglobin (Hb) concentration, micronutrient status (plasma ferritin, soluble transferrin receptor, retinol‐binding protein (RBP), folate, and vitamin B12), and the presence of Hb variants (Hb A, A2, S, and F; by HPLC). The prevalence of anemia, malaria, and inflammation was 45%, 8%, and 46%, respectively. Five children (1.6%) had sickle cell disease (HbSS), and 14% were Hb S carriers (HbAS). Anemia (Hb < 110 g/L) was more common among HbAS children compared to HbAA children (60 vs 42%, P < 0.05), but mean Hb concentrations did not differ (P=0.24). Inflammation‐adjusted plasma ferritin and RBP concentrations did not differ between HbAS vs HbAA children. Among children with anemia, 21% had 1 or 2 Hb S alleles; of the remaining HbAA children with anemia, 47% had malaria, low ferritin, and/or low RBP. The Hb S variant is prevalent among children in urban Cameroon and may explain a moderate proportion of anemia in this population. However, efforts are still needed to control anemia related to micronutrient deficiencies and infections.
Socioeconomic Inequalities in the Childhood Dual Burden of Malnutrition
Authors: Tzioumis Emma , Poole Charles , Bentley Margaret , Adair Linda
Published: June 2021

Evidence from adults suggests overweight is shifting to lower income groups in low‐ and middle‐income countries (LMIC), but has not been studied in children.

Using Demographic and Health Surveys from 28 LMIC with repeated surveys from 1990‐2012, we estimated country‐level prevalence of stunting (ST) and overweight (OW) in children <5y, and the prevalence difference of ST and OW between lowest and highest quintiles of household wealth within each country‐year. To determine if the disparity between lower and higher wealth changed over time, we calculated the annualized difference in prevalence growth rate for ST and OW from first to final survey within each country.

ST prevalence is greatest in the lowest wealth quintile, as indicated by positive prevalence difference for all country‐years (range: 0.9‐51.2). OW prevalence differences are mostly negative (range: ‐13.1‐3.3), indicating higher absolute prevalence of OW in the highest wealth quintile than the lowest in most, but not all countries.

The annualized difference in ST prevalence growth rate between quintiles was positive for 50% of countries, meaning the ST rate increased more quickly for the lowest wealth quintile than the highest in 50% of countries but increased more quickly in the highest wealth quintile in the other 50%. The annualized difference in OW prevalence growth rate between quintiles was positive for 57% of countries, indicating that OW prevalence increased more quickly in the lowest wealth quintile than the highest in a majority of countries.

Stunting remains the burden of the poorest children. Absolute overweight prevalence is still highest in the wealthiest children, but this is starting to shift to lower wealth groups.