bluuebunny/crossref_metadata_2025 huggingface dataset:
Men and women in space: bone loss and kidney stone risk after long‐duration space flight (257.3)
Authors: Smith Scott , Zwart Sara , Heer Martina , Hudson Edgar , Shackelford Linda , Morgan Jennifer
Published: June 2021

Bone loss on Earth is more prevalent in women than men, leading to the assumption that women may be at greater risk from bone loss during flight. Until recently, the number of women having flown long‐duration missions was too small to allow any type of statistical analysis. We report here data from 42 astronauts on long‐duration missions to the International Space Station, 33 men and 9 women. Bone mineral density (dual‐energy X‐ray absorptiometry), bone biochemistry (from blood and urine samples), and renal stone risk factors were evaluated before and after flight. Data were analyzed in two groups, based on available resistance exercise equipment. The response of bone mineral density to flight was the same for men and women, and the typical decrease in bone mineral density (whole body and/or regional) after flight was not observed for either sex for those using an Advanced Resistive Exercise Device. Bone biochemistry, specifically markers of formation and resorption, generally responded similarly in male and female astronauts. The response of urinary supersaturation risk to space flight was not significantly different between men and women, although risks were typically increased after flight in both groups and risks were generally greater in men than in women before and after flight. Overall, the bone and renal stone responses of men and women to space flight were not different.

Grant Funding Source: Funded by the NASA Human Research Program.
Phloridzin fatty acid esters induce apoptosis and alters gene expression in human liver cancer cells (261.2)
Authors: Nair Sandhya , Ziaullah Ziaullah , Rupasinghe H.P. Vasantha
Published: June 2021

Phloridzin Fatty Acid Esters Induce Apoptosis and Alters Gene Expression in Human Liver Cancer Cells Sandhya VG Nair, Ziaullah, HP Vasantha Rupasinghe*Department of Environmental Sciences, Faculty of Agriculture, Dalhousie University, Truro, NS B2N 5E3, Canada Abstract Phloridzin (phloretin 2‐O‐glucoside) is a dihydrochalcone, which is specifically found in apple and well known for its beneficial biological effects. To date, there is no report of antiproliferative effect of phloridzin on human cancer cell lines. We have synthesized novel acylated derivatives of phloridzin with six different long chain saturated, mono‐, and poly‐unsaturated fatty acids by regioselective enzymatic acylation. We investigated the effect of these six phloridzin fatty acid esters and their parent compounds on cell proliferation of hepatocellular carcinoma cells (HepG2) and its potential molecular mechanisms. Phloridzin fatty acid esters inhibited HepG2 cells growth in a concentration‐dependent manner by inducing apoptosis, as evidenced by decreased cell viability, formation of apoptotic bodies, decreased mitochondrial membrane potential, activation of caspase 3 and inhibition of DNA topoisomerases IIα activity that might induce G0/G1 phase arrest. However, phloridzin fatty acid esters did not significantly affect the growth of normal hepatocytes. Antiproliferative effect of phloridzin docosahexaenoic acid (DHA) ester was also associated with the down regulation of anti‐apoptotic genes, growth factor receptors, PI3k/AKT/mTOR as well as epigenetics regulators. Results reveal that phloridzin fatty acid esters possess potential as therapeutic agents for treating hepatocellular carcinoma.
Assessing an infant food frequency questionnaire: a pilot study (36.2)
Authors: Palacios Cristina , Torres Roxana , Trak Maria , Joshipura Kaumudi , Willett Walter
Published: June 2021

Adequate nutrition early in life optimizes growth and reduces the risk of obesity and chronic diseases in infancy and later in life. However, there is lack of data and instruments specifically designed to assess consumption early in life. This pilot study assessed an infant food frequency questionnaire (FFQ) for clarity of questions and to identify additional questions/examples for inclusion. This FFQ was based on the 19‐item FFQ developed by the CDC, which was expanded to 55 items with additional questions on preparation/source of foods and supplements. A convenience sample of mothers 蠅21 years with infants <12 months (m) of age attending the pediatric hospital clinics in San Juan, Puerto Rico were recruited. Sixty mothers (28 with infants 1‐4m; 14 with infants 5‐8m and 18 with infants 9‐12m) completed the FFQ. Most considered it to be clear (90%). Ten percent was not clear when to report frequency by day or week. The word baby food was more accepted than commercial baby pure. In the open‐ended options, the most common fruits and vegetables reported were bananas, carrots, green beans, squash and pears. In terms of frequencies, only 18% of infants at 1‐4m, 7% at 5‐8m and 6% at 9‐12m were exclusively breastfed. In combination with formula, 11% at 1‐4m, 7% at 5‐8m and 0% at 9‐12m were breastfed. Cow’s milk was introduced <12m in 11%. Water was introduced early (39% of infants at 1‐4 m). Fruit juice (any type) was consumed by 14% at 1‐4m, 57% at 5‐8m and 89% at 9‐12m old. “Baby juice” (juice marketed for babies) was consumed by 11% at 1‐4m, 50% at 5‐8m and 61% at 9‐12m. Sodas were not consumed. Solids foods (cereals, fruits & vegetables) were started <6m in 11%. Sweets were consumed by 12% starting at 7m. Most common supplements given were iron (67%) and multi‐vitamin (38%). Further work to assess validity of the FFQ is needed. A valid infant FFQ is important for understanding dietary consumption patterns in infants, which can help develop specific recommendations for fostering healthy eating habits in this stage of life.

Grant Funding Source: This project was supported in part by NIMHHD 8G12‐MD007600
Regulation of epidermal growth factor receptor signaling by clathrin‐coated membrane microdomains (603.2)
Authors: Antonescu Costin , Garay Camilo , Delos Santos Ralph Christian , Judge Gurjeet , Bautista Stephen , Lucarelli Stefanie , Pandey Rohan
Published: June 2021

Epidermal Growth Factor (EGF) binding to its receptor (EGFR) activates several signaling intermediates including Akt, leading to control of cell survival and metabolism. Concomitantly, ligand‐bound EGFR is rapidly incorporated into clathrin‐coated pits, eventually leading to receptor internalization. Whether clathrin scaffolds have functions at the cell surface prior to internalization, such as facilitating EGFR signaling, is poorly understood. To determine if clathrin scaffolds are required for activation of specific EGFR signaling pathways, we employed selective perturbation of clathrin (which prevents formation of clathrin scaffolds) and dynamin (which allows formation of clathrin scaffolds but prevents EGFR endocytosis). We find that siRNA gene silencing or pharmacological inhibition of clathrin, but not of dynamin, inhibits activation of the Akt pathway by EGFR but not receptor autophosphorylation. Using total internal reflection fluorescence microscopy, we have examined the localization of EGFR signaling intermediates vis‐à‐vis clathrin‐coated pits. Our findings indicate that clathrin scaffolds may represent unique signaling microdomains within the plasma membrane, a function that can be separated from the role of clathrin in endocytosis.

Grant Funding Source: Supported by the Canadian Institutes of Health Research
Transcriptional regulation of O‐GlcNAc homeostasis (607.8)
Authors: Qian Kevin , Ruan Hai‐Bin , Yang Xiaoyong
Published: June 2021

A vast number of nuclear and cytoplasmic proteins are modified by O‐linked β‐N‐acetylglucosamine (O‐GlcNAc), a dynamic post‐translational modification that has emerged as a key regulator of diverse cellular processes. The addition and removal of O‐GlcNAc are catalyzed by O‐GlcNAc transferase (OGT) and O‐GlcNAcase (OGA), respectively. Accumulating evidence indicates that both excess and deficiency of O‐GlcNAc in the cell can have deleterious effects, suggesting that maintenance of O‐GlcNAc homeostasis is critical for cellular function. However, the mechanism by which O‐GlcNAc homeostasis is maintained in the normal state and disrupted in the disease state has not yet been investigated. Here, we demonstrate that Ogt expression is upregulated by OGA, creating a feedback mechanism that serves to maintain a balance between OGT and OGA expression levels in the cell. We found that OGA promotes Ogt transcription through cooperation with the histone acetyltransferase p300 and the transcription factor CCAAT‐enhancer‐binding protein β (C/EBP‐β). In this protein complex, OGA and p300 act as transcriptional coactivators of C/EBP‐β through their glycosidase and acetyltransferase activities, respectively. Since O‐GlcNAc homeostasis is often disrupted in cancer cells, we also sought to determine whether classical oncoproteins are involved in regulating Ogt expression. We found that extracellular signal‐regulated kinase (ERK) increases OGA glycosidase activity and promotes Ogt transcription, suggesting that chronic growth factor stimulation may be responsible for disrupted O‐GlcNAc homeostasis in cancer. Our study elucidates a transcriptional mechanism that controls cellular O‐GlcNAc homeostasis, which should lay a foundation for exploring O‐GlcNAc signaling as a therapeutic target for human disease.

Grant Funding Source: Supported by NIH R01‐DK089098
High school mentors' perceptions about Veggiecation within HealthCorps: a case study (624.31)
Authors: Bai Yeon , Birnbaum Amanda , Fly Alyce
Published: June 2021

New and creative mechanisms are needed to combat childhood obesity. HealthCorps®, a nonprofit organization devoted to improving health, is staffed by recent college graduates to promote healthy eating and physical activity to high school students. This study explored HealthCorps®‐trained high school mentors’ perceptions, i.e. benefits, challenges, and perceived success of Veggiecation®, a nutrition education program composed of cooking, tasting and other interactive activities. Trained high school students in Bronx led Veggiecation® in an afterschool program with 30 students from 2 middle schools, 2‐4 sessions/month for 7 months during 2012‐13. Results showed that high school mentors identified personal benefits, including exposure to new vegetables, enjoyment of recipes and tasting, and learning about healthy eating. Mentors also reported learning how to teach younger teens and personal satisfaction as other benefits. Reported challenges were mostly external, related to space/logistics, engagement of middle school students in lessons, and persuading them to taste recipes. All peer mentors reported that Veggiecation® was successful in improving children’s vegetable intake, citing recipes and tasting as the most successful components. This approach, which mobilized existing resources and could be expanded, should first be studied for effectiveness to both mentors and program participants.
A study on high school students’ recognition of food additives in Incheon region, Korea (626.3)
Authors: Kim Young A , Kim Eun‐ju
Published: June 2021

This study was conducted to provide basic information about how high school students recognize food additives in Incheon region, 5th largest city in Korea, using a questionnaire and data analysis tool, SPSSWIN 18.0.

50% of students were found that they know about food additives. They recognized the harmfulness of food additives because that it was synthesized chemicals(59.6%, p<.05). But a test about 5 major food additives proved that they couldn’t’ distinguish between sweetener and seasoning (p<.05), and their interest about food additives except MSG was examined differently between the boys and girls. Only about 30% of students were likely to choose a food with less food additives. 71.5% of students answered that they would try to choose a food with natural additives in spite of higher cost, while 10% answered they surely choose a food with natural additives. The necessity of education about food additives was supported by 48% of girls and 35% of boys(p<.001), and the most popular reason of it was healthy and safe dietary life. As an educational method, education by a professional instructor was best supported by students, and there was a meaningful difference between boys(45.9%) and girls(54.5%), while the tentative attendance were replied by 31.9% of boys and 39.3% of girls(p<.01).

Therefore a professional education program to let students be aware of food additives for advancing their dietary life is required.

Grant Funding Source: Supported by Inha University
Nutrition knowledge scores of Korean male and female high school students (626.5)
Authors: Chang Kyung Ja , Lee Kyong Won
Published: June 2021

The purpose of this study was to examine nutrition knowledge of Korean high school students for nutrition education.The subjects were 276 high school students (male 138, female 138) in Incheon metropolitan city, Korea. A cross‐sectional survey was conducted using a questionnaire and the statistical data were analyzed with SPSS 12.0 program. As for the distribution of weight by BMI, there was significant difference between male and female students (p<0.001); 54.3% of the male normal, 10.9% overweight, 14.5% obese, 20.3% underweight and 47.8% of the female normal, 40.6% underweight, 12.2% obese, 9.4% overweight. As for nutrition knowledge scores, the female students (31.97/50) showed significantly higher average score compared to the male students (29.96/50) (p<0.05). There was no significant difference in nutrition knowledge scores according to BMI in the male students. However, there was significant difference in nutrition knowledge scores according to BMI in the female students (p<0.05); underweight (32.86/50), normal (32.50/50), overweight (27.69/50), obese (22.50/50) in order. As for source from which they got nutrition knowledge, there was no significant difference between male and female students. There was significant difference in nutrition knowledge scores according to source from which they got nutrition knowledge (p<0.05); dietitian (35.75/50), nutrition education class (32.35/50), mass media (31.37/50), internet (31.00/50), family, relatives and friend (28.25/50). Therefore, nutrition education according to gender and BMI should be provided in high school in Korea.
Muscle mass as a potential predictor for metabolic syndrome in normal weight adults (628.20)
Authors: Gee David , Humphrey Julia , Williams Daniel , Hawk Susan
Published: June 2021

The prevalence of abnormal metabolic conditions in normal weight subjects has been recognized, but the cause is unknown. However it has been proposed that a decrease in relative muscle mass may be a contributing factor. The purpose of this study was to assess the relationship between appendicular lean mass (ALM) index (kg/m2) and risk factors of metabolic syndrome (MetS) in normal weight adults using data from the continuous National Health and Nutrition Examination Survey years 1999‐2006 (n = 1826). Chi‐square analysis and odds ratios were used to establish significant differences in prevalence and likelihood of MetS risk factors. The prevalence of MetS, hyperglycemia, hypertension, and hypertriglyceridemia were significantly higher in both sarcopenic (lowest tertile ALM index) males and females; however, this relationship was not as strong when the population was stratified into age groups. Increased risk for hypertension was significant in sarcopenic males in the 60+ age category (P = 0.0076) and for hypertriglyceridemia (P = 0.0240) and MetS (P = 0.0026) in sarcopenic females in the 40‐59 age category. Increased prevalence for MetS and its risk factor components were consistently observed with age‐related reductions in relative muscle mass. These results suggest the importance of recognizing decreased relative muscle mass as a risk factor for MetS in normal weight middle‐aged and older adults.
Performance of young and aged C57BL/6J mice on cognitive task depends on the task (629.2)
Authors: Du Kristy , Perez Samuel , Rhodes Justin
Published: June 2021

Aging has been associated with weakening of cognitive function, and nutritional supplementation has been suggested to enhance cognitive function. Prior research has demonstrated limited effects of dietary supplementation on rodent performance on cognitive tasks. The objective of this study is to identify behavior tasks that provide the greatest degree of sensitivity in aging‐associated cognitive decline for use in future studies. Young (2 months old) and aged (18 months old) female and male C57BL/6J mice were evaluated for performance on the Morris water maze, elevated plus maze, novel object recognition, passive avoidance, and active avoidance. Animals were euthanized at the end of the study to quantify proliferation of new neurons using doublecortin immunohistochemistry. The largest statistical differences in performance between age groups were seen in novel object recognition and active avoidance. This research model will lay the foundation for understanding the underlying mechanisms by which nutrition interventions may be implemented to slow the cognitive decline associated with aging.
Eicosapentanoic acid increases aquaporin‐3 expression in human skin keratinocytes (647.49)
Authors: Mun Yeun‐Ja , Jeon Byoung‐Kook , Woo Won‐Hong , Lee Young‐Eun
Published: June 2021

Eicosapentanoic acid (EPA), an omega‐3 polyunsaturated fatty acids (ω‐3 PUFAs), protects against photodamage and photocarcinogenesis in mammals. Water movement across the plasma membrane can occur via two pathways: by diffusion through the lipid bilayer and by membrane inserted water channels (aquaporins; AQPs). At least 13 subtypes, denoted AQP0‐AQP12, have so far been identified in mammalian cells. AQP3 is a water/glycerol transporting protein in basal layer keratinocytes of epidermis in normal skin. We have investigated the effects of EPA on AQP3 expression and the effects of EPA on ultraviolet (UV)‐induced AQP3 down˗regulation in cultured human skin keratinocytes. EPA treatment increased AQP3 gene and protein expression in human epidermal keratinocytes (HaCaT). Using a specific inhibitor, we observed that the effect of EPA on AQP3 expression was mediated by ERK activation. UV induced AQP3 down‐regulation in HaCaT cells. EPA treatment attenuated UV‐induced AQP3 loss and cell death. Collectively, the present results show that EPA increased AQP3 expression and protected against photodamage of UV through the induction of AQP3 expression.
Niclosamide blocks autophagy via lysosomal dysfunction (663.18)
Authors: Gao Ying , Liang Aiwen , Kang Jeong‐Han , Liu Peiqing , Yin Xiao‐ming , Li Min
Published: June 2021

Niclosamide is a salicylanilide derivative used as antihelminthic agent. Recently, Nic has been screened out as an autophagy modulator by us and other groups and was identified to be a structurally distinct mTORC1 signaling inhibitor. In this work, we measured the long‐lived protein degradation activity and found Niclosamide could inhibit the autophagic process. To elucidate the inhibition of Niclosamide on autophagy, we further tested the lysosomal activity using DQ‐BSA assay and measured protease activity such as Cathepsin B and Cathepsin D with fluorescent substrates. The lower degradation of DQ‐BSA and decreased protease activity after Niclosamide treatments indicated the impaired lysosome function. In addition, both LC‐II and P62 proteins accumulated significantly when treated by Niclosamide suggesting the late stage of autophagy was blocked. The increased pH of lysosome and release of Cathepsins from lysosome to cytosol demonstrated that Niclosamide might affect lysosome resulted from lysosomal membrane permeabilization. Collectively, we firstly disclosed Niclosamide not only inhibited mTORC1 activity but also, as other well‐known lysosome inhibitors, strongly blocked autophagy via lysosomal dysfunction.

Grant Funding Source: Supported by Guandong Provincial Natural Science Foundation, China (S2013010015876)
Severity of behavioral impairments and vascular dysfunction with chronic stress/depressive symptoms is increased by metabolic syndrome (676.7)
Authors: Brooks Steven , Stanley Shyla , Butcher Joshua , D'Audiffret Alexandre , Skaff Paulina , Chantler Paul , Frisbee Jefferson
Published: June 2021

Chronic stress/depressive symptoms impair the normal patterns of vascular reactivity. However, they also can increase CVD risk owing to an elevation in markers of the metabolic syndrome (e.g., blood pressure, glycemic control, dyslipidemia). To interrogate this, we imposed 8 weeks of unpredictable chronic mild stress (UCMS) onto lean (LZR) and obese (OZR) Zucker rats to determine their behavioral and vascular outcomes. UMCS resulted in depressive symptoms in LZR, including reduced chronic and stimulated grooming, learned helplessness, and increased plasma cortisol. A directionally consistent, although milder, effect was determined in control OZR. In ex vivo aortic rings and skeletal muscle arterioles, endothelium‐dependent dilation was blunted by either metabolic syndrome (OZR) or imposition of UCMS (LZR) owing to increased ROS and TxA2 production. However, when OZR were exposed to UCMS, the severity of the depressive symptoms was increased, and dilator reactivity in conduit/resistance vessels was nearly abolished. These observations suggest that a combination of metabolic syndrome and depressive symptoms creates a condition causing rapid degradation of vascular function.

Grant Funding Source: Supported by National Institutes of Health and American Heart Association
NF‐κB inhibitor MG132 enhances differentiation and collagen expression of dental pulp stem cells (732.1)
Authors: Saffarian Tousi Hozhabri Neda , Kim Harry , Varanasi Venu
Published: June 2021

Inflammatory response in the dental pulp is mediated by interleukin‐1β and tumor necrosis factor‐α, which may alter collagen matrix formation by dental pulp stem cells and lead to a delay or poor healing of the dental pulp. We hypothesized that these inflammatory cytokines inhibit dental pulp stem cell collagen production by activating the transcription factor nuclear factor ‐ κB. The purpose of this study is to determine the role of nuclear factor ‐ κB signaling during odontoblastic differentiation of human dental pulp stem cells and collagen synthesis under inflammatory culture conditions. Data for the study were obtained by qRT‐PCR and Picrosirius staining for gene expression and collagen matrix staining, respectively. Scanning electron microscopy was used for generating high‐resolution images of the samples. Results showed a significant increase in nuclear factor ‐ κB gene expression by interleukin‐1β and tumor necrosis factor‐α, and this was inhibited in a dose‐dependent manner by a nuclear factor ‐ κB inhibitor (MG132). Interestingly, dental pulp stem cells exposed to inflammatory conditions resulted in a significant increase in collagen (I)‐α1 gene expression, which was further enhanced in the presence of the inhibitor. This correlated with collagen matrix staining. Nuclear factor ‐ κB inhibition also enhanced the gene expression of the odontoblastic marker dentin sialophosphoprotein and demonstrated an odonotblastic cell morphology for the promotion of an odontogenic lineage during inflammation. Conversely, inhibition of NF‐kB in the absence of inflammation resulted in an increase in the gene expression of osteogenic markers (runx2, osteocalcin). In conclusion, controlling inflammatory signaling in DPSCs by reducing the activity of NF‐kB enhances odontoblastic differentiation and collagen matrix formation, potentially stimulating pulp and dentin healing.

Grant Funding Source: NIH grants #A107380, #1K25DE018230‐01, #5P30DE020742‐02
Saline stress modulates mir‐27a, mir‐103 and mir135 in 3T3‐PKD‐1 cells (802.15)
Authors: Torres‐Marquez M Eugenia , Ruiz Esparza‐Garrido Ruth , Sinnett‐Smith Jim , Rozengurt Enrique
Published: June 2021

PKD is a founding member of CAMK group of kinases and there is ample evidence of its participation in important cellular process including stress. Due to the lack of specific inhibitors for its action in vivo, its protein targets are scarcely described. Even less is known about its role at regulating the transcription of target proteins. The miRNA are important regulators of protein expression and several groups are currently undertaking studies about its targets, enriching a database that will enable a better understanding of the networks in which they participate.

In this work we aimed to identify miRNA that are modulated by hyperosmotic stress in 3T3 expressing PKD‐1 cells as a means to understand the PKD participation in this cellular process.

We obtained the RNA from 3T3‐PKD‐1 cells and their control counterparts, without or with an hour challenge with 0.3M NaCl, and sought to identify the up‐ or down‐ regulated miRNA. This was performed using miRNA3.0 Affymetrix microchips, and analyzed with software provided.

We found that miRNA that are exclusively regulated by the PKD action after the 0.3M NaCl challenge are: mir‐103, mir135, which are down‐regulated, and mir27a, which is up‐regulated.

Some of the targets of the down regulated mir‐103 include Syntaxin‐17, which participates in the autophagosomes formation; inhibitory growth factor 5 which participates in proliferation; and cadherin‐11, which participates in shaping cell morphology. The mir‐135 has as targets complexin 1 that participates in vesicle transport. The up‐regulated mir‐27a has as targets the A2B adenosine receptors that help repair ischemia/reperfusion injury, the activin A receptor ‐type C and the Ubiquitin conjugating enzyme E2N and Polo‐like kinase that regulates the differentiation of various organs.

Grant Funding Source: IN222014
Supplementation with multiple micronutrients and polyunsaturated fatty acids does not improve school performance of obese children in a rural area of Mexico (811.23)
Authors: Escobedo Sara , García Olga , Ronquillo Dolores , Caamaño María , Rosado Jorge
Published: June 2021

The study investigated the effect of nutrition education and supplementation with multiple micronutrients (MM) and polyunsaturated fatty acids (PUFA) in the school performance of obese children in a rural area of Queretaro, Mexico. In a longitudinal, randomized, controlled trial, 134 children (6‐10 y) were assigned to one of two groups: a) 237mL/day of a supplement with MM and PUFA’s (SMP) and nutrition education; b) nutrition education alone (CON). All treatments were administered for 24 wks. Weight, height, waist circumference and body composition (by DXA) were measured at 0 and 24 wks. Diet was evaluated at the beginning and at the end of the study using 3 of 24h recalls. School performance was evaluated with school grades before and after the intervention, and with the CONNERS questionnaire applied to parents and teachers at 24 wks. Vitamins A and C intake at baseline was positively associated with Spanish, mathematics and overall grades (p<0.05). There were no differences between groups in school performance at the end of the study. Also, no significant differences between groups were observed in the CONNERS evaluation of teachers or parents. In conclusion, nutrition education and supplementation with MM and PUFA’s does not improve school performance of obese children living in rural Mexico.

Grant Funding Source: Supported by CONACYT
An enhanced photonic crystal biosensor‐based immunodiagnostic assay for detection of soluble transferrin receptor as a biomarker of iron deficiency (813.7)
Authors: Peterson Ross , Cunningham Brian , Andrade Juan
Published: June 2021

Identification of populations at risk for iron deficiency is of critical concern. Studies on the detection of soluble transferrin receptor (sTfR) using a PC biosensor assay are presented. sTfR (85kDa) is a favorable biomarker of iron status as it is less confounded by inflammation related factors. PCs are periodic dielectric surface structures designed to reflect a narrow band of wavelengths when illuminated by a broadband light source. Adsorption and binding of biomolecules on the PC surface elicits a wavelength shift (PWV) measured in nm. The PC surface of 384‐well microplate was functionalized with silane for immobilization of capture antibodies (Ab), followed by a blocking step. sTfR antigen was added before secondary Ab was dispensed in standard and control wells. Secondary Ab was functionalized with iron‐oxide nanoparticles (30 nm) to enhance signal‐to‐noise ratio. Antigen‐Ab interactions were followed using the BIND system. A sTfR standard curve (200‐5 ng/ml) was created and compared against a commercial ELISA kit. Ab selectivity, limit of detection (LOD), and dose‐response were evaluated (two experiments, n=4/sample). The PC biosensor showed a LOD of 28 ng/ml, a linear response ([PWV=0.8 + 8.4(ug/ml)‐1]; R2=0.98) and limited non‐specific binding against BSA. This study demonstrates the feasibility of the PC biosensor as an immunodiagnostic assay for sTfR detection.

Grant Funding Source: ACES VISION 20/20 Grant
Effects of rice bran oil deodorizer distillate on plasma cholesterol levels in rats (829.23)
Authors: Gomes Tina , Gu Xuan , Min Byungjick , Lee Sun‐Ok
Published: June 2021

Rice bran oil deodorizer distillate (RBODD) is a byproduct in the refinement of rice oil. RBODD has received great interest as a potential health‐promoting product because it is a rich source of tocopherols and tocotrienols. Hypercholesterolemia is one of the major risk factors for coronary heart disease. The objective of this study was to investigate the effect of RBODD on plasma cholesterol concentrations in Sprague Dawley rats. Thirty three 6 week old rats were randomly divided into three groups: normal fat control (NFC), high fat control (HFC) and high fat diet plus RBODD treatment (RBO). 45 mg of RBODD was orally administered daily for 3 weeks into each treatment rat using 5% of Tween‐80 as a vehicle. The rats in the control groups were received Tween‐80 alone in the same manner. At the end of the feeding period, blood samples were collected by cardiac puncture for lipid analysis. RBO group had significantly lower plasma triglyceride compared with HFC group (p<0.05). RBODD treatment did not show changes in the cholesterol levels. This study suggests that the efficacy of RBODD on plasma lipid profile is to improve triglyceride levels.

Grant Funding Source: Supported by Arkansas Bioscience Institute (ABI)
Nanocurcumin attenuates inflammation by decreasing Toll‐like receptor 2 and 4 expression and activity and promoting an anti‐inflammatory macrophage phenotype (830.22)
Authors: Chen Xinpu , Chenna Venugopal , Maitra Anirban , Devaraj Sridevi
Published: June 2021

Curcumin, the yellow polyphenol extracted from turmeric (Curcuma longa), has potent anti‐inflammatory properties. However, due to poor aqueous solubility and thus limited systemic bioavailability, it has not been used clinically. Nanoparticle‐based drug delivery approaches have the potential for rendering hydrophobic agents like curcumin dispersible in aqueous media. We have synthesized a polymeric nanoparticle encapsulated curcumin ‐ nanocurcumin (NC) and have shown that it is readily dispersed in aqueous media and demonstrates comparable in vitro therapeutic efficacy to free curcumin. Obesity and diabetes are pro‐inflammatory states characterized by increased monocyte Toll like receptors (TLR) 2 and 4 and a propensity to a M1 pro‐inflammatory phenotype. In this study, we aimed to examine the effects of NC on monocyte TLR2 and 4 expression and activity and on macrophage phenotype and subsequent inflammatory pathways. Incubation of monocytes with NC (5uM) compared to vehicle control resulted in 50% inhibition of TLR2 and 60% inhibition of TLR4 expression, a 35% reduction in downstream signaling, ie. MyD88 expression and NFKb activity. Furthermore, incubation with NC resulted in conversion of macrophages from M1 to M2 phenotype. All of these beneficial effects of NC led to a significant downregulation of several pro‐inflammatory cytokines and chemokines (IL‐6, IL‐8, MCP‐1 and TNFα). These studies suggest that NC could be a potent adjunctive anti‐inflammatory therapeutic strategy to consider in vivo, especially in obesity and diabetes.
Ferritin induces IL‐1β production through inflammasome activation via NF‐κB‐dependent manner in macrophages (835.3)
Authors: Ikeda Yasumasa , Tajima Soichiro , Imao Mizuki , Horinouchi Yuya , Izawa‐Ishizawa Yuki , Kihira Yoshitaka , Ishizawa Keisuke , Tsuchiya Koichiro , Tamaki Toshiaki
Published: June 2021

Ferritin (Ftn), an iron‐storage protein, is widely used as an indicator of body iron storage. Serum ferritin was upregulated under the inflammatory status. Recently, the secreted Ftn exerts certain biological actions, however, it remains unclear whether the secreted Ftn itself promotes inflammation. Here, we investigated the biological action of Ftn on inflammation. Murine peritoneal ascites‐derived macrophage, RAW264.7, was used in this study. Exogenous Ftn treatment as well as iron‐free Apo‐Ftn induced IL‐1ß expression and maturation in macrophage, resulting in increased IL‐1ß secretion in the supernatant. Ftn and Apo‐Ftn promoted caspase‐1 activation and upregulated the expression of nucleotide‐binding oligomerization domain (NOD)‐like receptor family, pyrin domain‐containing 3 (NLRP3), inflammasome activation. Ftn‐induced IL‐1ß activation and secretion were inhibited by caspase‐1 inhibitor or NLRP3 silencing. Ftn‐induced inflammasome activation was independent of iron content and inhibited by the pharmacological inhibition of nuclear factor kappa B (NF‐κB). Thus, the secreted Ftn possess proinflammatory activities to promote IL‐1ß production through the NF‐κB‐dependent transcriptional regulation of NLRP3‐inflammasome in macrophages, which is a novel finding of ferritin beyond the indicator of body iron content.
Cell‐synthesized extracellular matrix particles for tissue engineering and aesthetic applications (87.2)
Authors: L'Heureux Nicolas , Young D , Dusserre Nathalie , Sonnenberg Sonya , Johnson Todd , McAllister Todd , Rodriguez Ricardo , Christman Karen
Published: June 2021

With age, or after injury, subcutaneous adipose and conjunctive tissue loss can cause poor aesthetic appearance. Current injectable tissue fillers trigger foreign body responses that eventually lead to their degradation or encapsulation. The ideal dermal filler would provide initial structural support but also trigger a regenerative process and lead to the creation of a new permanent tissue. The goal of this project is to develop a new class of fillers that provides long‐term results by triggering the regeneration of a stable, living and vascularized adipose tissue.

Using the Tissue Engineering by Self‐Assembly (TESA) approach, sheets of ECM produced by normal human fibroblasts in vitro were dehydrated and reduced to a particulate form that could be injected subcutaneously in nude mice. The ECM particles caused minimal inflammation or encapsulations. Most notably, vascularization and adipogenesis were observed at the injection site, neither of which was noted when using two popular dermal fillers (Juvederm® or Radiesse®). Progression of adipogenesis around the ECM particles was noted throughout the duration of the study (3 months). In separate studies, particles were cast in vitro to produce finely channelized structures that could be seeded with endothelial cells. This approach offers a viable strategy for the production pre‐vascularized bulk tissue to address secretory organ tissue engineering.

Conclusion: Fibroblast‐synthesized particles combine “off‐the‐shelf” availability and a persistent ECM that provides mechanical support while triggering the development of a vascularized adipose tissue. Clinical studies are underway to compare this new regenerative dermal filler with the standard of care.
Biocompatibility of halloysite clay nanotubes in a rat dermal model (87.4)
Authors: Mills David
Published: June 2021

The cytocompatibility of halloysite nanotubes (HNTs) have been firmly established. The biocompatibility of HNTs has yet to be established. The host‐immune response to dermally implanted HNTs in a rat skin wound model was assessed over a one‐month experimental period. Adult Sprague Dawley rats (27, male, 300g) comprised the study population. A calibrated microinjector was used to inject a 25μl PBS solution, plus/minus HNTs. Control group I animals received implant site preparation and microneedle penetration only while Control group II animals received implant site preparation, microneedle penetration, and a PBS injection. Clear bandages were applied over each implant site permitting daily visual observations. Animals were monitored and photodocumented on a daily basis for changes in coloration, infection, and healing pattern. Animals (n=3, each group) were sacrificed at one, two and four week intervals. Tissue blocks, containing implant sites and representing full thickness skin tissue sections, were harvested, fixed in 4% buffered formalin, paraffin embedded and sectioned at six microns. Histochemical and Immunohistochemical analysis were used to assess the host immune response. All analyses showed that implanted HNTs did not provoke either a local or systemic immune response. As HNTs are proving to be a potent drug delivery vehicle, our data provides support for their future use in clinical medicine.

Grant Funding Source: Governor's Biotechnology Initiative
An animated 3D model of the synchronous movements of the suprasyringeal structures and organs in the neck of a vocalizing songbird, the Northern Cardinal (Cardinalis cardinalis) (918.8)
Authors: Blevins Caroline , Ge Jinghua , Suthers Roderick , Homberger Dominique
Published: June 2021

In birds, sound is generated in the syrinx and is filtered by suprasyringeal structures that move relative to one another. The larynx moves relative to the hyoid apparatus, which moves relative to the mandible, which in turn moves relative to the skull. The cervical vertebral column moves the head relative to the thorax. The esophagus and trachea move relative to the vertebral column. The interplay of these elements was analyzed in a songbird, the Northern Cardinal, through an animated 3D model that was created from a CT scan based on prior anatomical analyses. The movements of the vocalizing behavior were modeled by matching the configuration of the 3D model to individual images of a 2D x‐ray video of a singing bird in lateral view. To estimate the degree of yaw and roll of the head, the x‐ray video was synchronized with a video recorded from above. To compensate for side‐to‐side movements of the bird, the images of the 2D x‐ray video were enlarged or reduced. A separately recorded x‐ray video in frontal view was used to verify the proper spatial configuration of the skeletal elements in the individual images of the animated 3D video. The video of the 3D model was synchronized with the audio recording to analyze the movements required to produce specific sounds.

Grant Funding Source: Supported by NIH grant NINDS R01 NS029467 to RAS; LSU Foundation to DGH
A new approach to study site‐specific protein sumoylation (925.3)
Authors: Albuquerque Claudio , Yeung Eyan , Ma Shawn , Fu Ting , Zhou Huilin
Published: June 2021

Protein sumoylation is a reversible post‐translational modification that regulates nuclear processes including gene transcription, nuclear transport and DNA repair. Like ubiquitin, SUMO is attached to the lysine side chains of target proteins via a cascade of E1‐activating, E2‐conjugating, and E3‐ligating enzymes, and it is removed by SUMO‐specific isopeptidases. Hundreds of proteins are known to be sumoylated in the yeast saccharomyces cerevisiae, however, relatively little is known about the precise sumoylation site(s) of most proteins. This is due to a lack of suitable method to map sumoylation sites in vivo. As a result, the function of sumoylation has remained mysterious in most cases. We have recently identified a novel role of protein sumoylation in preventing genome rearrangements and developed proteome‐wide approach to identify and quantify sumoylated proteins. Here we will report a new proteome‐wide method for the identification of site‐specific sumoylation in yeast, which greatly expanded the current knowledge of protein sumoylation. In addition, we will describe new approaches for the analysis of protein sumoylation in vitro. Results will be presented on the functional analysis of sumyolation of the sumoylation enzymes in yeast.

Grant Funding Source: LICR
Unnatural covalent bond formation inside and between proteins through proximity‐enhanced reaction (972.6)
Authors: Xiang Zheng , Ren Haiyan , Hu Ying , Coin Irene , Wei Jing , Cang Hu , Wang Lei
Published: June 2021

The cysteine disulfide bond plays a crucial role as intramolecular linkage in protein folding, stability and function. However, its inherent redox sensitivity and reversibility limit its applications in biotechnology and therapeutics. We envisioned that a new irreversible and stable covalent bond formed between a genetically incorporated unnatural amino acid (Uaa) and the residue of a natural amino acid would complement our ability of protein engineering and provide new tools for biological studies. We genetically incorporated a new electrophilic Uaa, p‐2’‐fluoroacetylphenylalanine (Ffact), into different proteins in E. coli and mammalian cells using the orthogonal tRNA‐synthetase pair, and demonstrated its applications in three ways. First, the incorporation of Ffact into the Z protein affibody at a specific position enabled it to form a covalent bond with Z protein mutant with Cys mutation. The reaction was enhanced by the proximity of the Uaa and the Cys residue and did not happen between distal Uaa and Cys residue. Second, we incorporated Ffact into the corticotropin release factor receptor type 1 (CRF‐R1), a class B G protein‐coupled receptor (GPCR), at the ligand‐receptor binding region. Different urocortin‐1 (Ucn‐1) ligand analogs of CRF‐R1 with cysteine mutation were incubated individually with HEK293T cells expressing the Ffact‐CRF‐R1 mutant. The Ffact‐cysteine reaction happened site‐specifically. The data enabled us to further explore the mechanism of the class B GPCR activation. Third, we incorporated Ffact into fluorescent proteins (FP) to form intramolecular linkage between the fluorophore and the β‐barrel. The linked FP mutants showed a higher quantum yield and greater photon output compared to the unlinked FP mutants. These results indicated that it is possible to add a new covalent bond to proteins by incorporating Uaas with proximity‐enhanced reactivity, which will provide new tools for biological studies and protein engineering.

Grant Funding Source: Supported by CIRM (RN1‐00577‐1) and US NIH (1DP2OD004744‐01, P30CA014195)
Link between antioxidant effect and antiinflammatory response in butyrate arrested vascular smooth muscle cells (VSMC) proliferation (LB250)
Authors: Ranganna Kasturi , Mathew Omana , Milton Shirlette
Published: June 2021

Epigenetic mechanisms by altering the expression and in turn, functions of target genes, without changing their primary structure have potential to modify cellular processes that are characteristics of atherosclerosis such as inflammation, proliferation, migration and apoptosis/cell death. Butyrate, a natural epigenetic modifier and a histone deacetylase inhibitor (HDACi), is an inhibitor of VSMC proliferation, a critical event in the pathogenesis of atherosclerosis, and alters cellular ROS and glutathione levels suggesting involvement of redox‐component in antiproliferation effect of butyrate. Here the goal is to explore whether the antioxidant effect of butyrate contributes to antiproliferation action on VSMC via modulation of inflammatory response by using western blotting and immunostaining methods. The results show treatment of VSMC with butyrate not only upregulates glutathione peroxidase (GPx)‐3 and GPx‐4, but also increases overall catalytic activity of GPx further supporting antioxidant effect of butyrate in proliferation arrested VSMC. Moreover, analysis of NF‐kB transcription factor system, the target of GPx, reveals butyrate treatment of VSMC causes downregulation of p65NF‐kB, IkBα, IKKα and IKKβ expression and inhibits phosphorylation of p65NF‐kB resulting in downregulation of NF‐kB target inflammatory genes such as inducible nitric oxide synthase, vascular cell adhesion molecule and cyclooxygenase‐2. Overall these observations suggest a link between antioxidant effect and antiinflammatory response in butyrate arrested VSMC proliferation and emphasize the atheroprotective and therapeutic potential of natural products such as butyrate in vascular proliferative diseases.

Grant Funding Source: G12RR03045‐21 and C06RR012537 from NIH/NCRR
An enlarged model of intestinal villosity in papier‐mâché by Jérome AUZOUX, 19th century (LB3)
Authors: LE FLOCH‐PRIGENT Patrice , Gillot Jean‐Bernard , Barbet J. Patrick
Published: June 2021

An enlarged anatomical model of an intestinal villosity had been realized in papier‐maché by a french, medical doctor Jérome AUZOUX (19th century) who was specialized in human, animal and vegetal models of anatomy in this uncommon material which were enlarged and dismantabled. The villosity was about 30 cm high and was fixed on a rectangular, black, wooden basis with the label of Auzoux. Two main vessels : a small artery and a vein of the same caliber structured the sample on each side of a villosity shaped as a half cone. The sagittal plane was bordered by the drawing of the cells. Between the artery and the vein, numerous transverse anastomoses were represented. This model was quite simple in the production of Auzoux but it was a successful assay to represent a structure quite at an histological level of enlargement.
Consumption of frozen meals as compared to quick service restaurant meals is associated with better diet quality in adult participants of the National Health and Nutrition Examination Survey (2003‐2010) (LB383)
Authors: Papanikolaou Yanni , Comerford Kevin , Krumhar Kim , Fulgoni Victor
Published: June 2021

What We Eat In America (WWEIA)/NHANES (2003‐2010) data were used to determine associations between diet quality in adult (19+yrs) frozen meal (FM) consumers as compared to consumers of meals from quick service restaurants (QSR). Intake was determined using a 24 hour recall; FM consumers were defined as those who reported consumption of any of the 91 specific frozen meal codes while QSR meal consumers were those who reported obtaining meals from “restaurant fast food/pizza.” ANOVA and covariate adjusted means were determined using appropriate sample weights; significance was set at p<0.01. Diet quality was calculated using the Healthy Eating Index‐2010 (HEI), which is designed to measure conformance to the 2010 Dietary Guidelines for Americans. Compared to QSR consumers (n=6,238), FM consumers (n=287) had lower intakes of energy (2326±20 v 2073±51 kcal/d) and higher total HEI scores (53.0±1.5 v 44.4±0.4). FM consumers had higher HEI sub‐component scores for higher intakes of total vegetables (3.4±0.1 v 2.9±0.03), greens and beans (1.7±0.2 v 0.9±0.04), whole grains (2.9±0.3 v 1.8±0.1), total protein foods (4.4±0.1 v 4.1±0.03), and lower intakes of refined grains (6.6±0.4 v 5.4±0.1), and empty calories (13.1±0.5 v 9.9±0.2). Other HEI sub‐components (sodium, total fruit, whole fruit, dairy, seafood and plant protein, and fatty acid ratio) were not different among FM or QSR consumers. Frozen meal consumption was associated with lower energy intake and a better diet quality as compared to those consuming QSR meals.

Grant Funding Source: Supported by Nestlé