bluuebunny/crossref_metadata_2025 huggingface dataset:
Chronic and Acute Hypoxia Markedly Alter Ca2+ Signaling in Adult and Fetal Pulmonary Arterial Myocytes
Authors: Cothran Kyle , Sandy Scott , Romero Monica , Dobyns Abby , Paez Ricardo , Francis Michael , Taylor Mark S , Longo Lawrence D , Wilson Christopher G , Wilson Sean M
Published: June 2021

Ca2+ signaling is well regarded as being vital to hypoxic induced pulmonary arterial (PA) vasoconstriction and chronic high altitude living can exacerbate this process. To test the hypothesis that chronic hypoxia will increase Ca2+ wave activity due to acute hypoxia in pulmonary arterial myocytes (PAM) of both fetal and adult sheep and better understand the Ca2+ signals, we exposed pregnant and non‐pregnant ewes to 3800m for >110 days to induce chronic hypoxia. We then performed confocal imaging of PAM in Fluo‐4 loaded intact arteries using an en face preparation. Images were recorded in an atmospheric chamber at ~37°C adjusted to either normoxic conditions, at ~300m, or acute hypoxic conditions, simulated by exposing PAs to ~4% O2 for 30 min. Ca2+ signal characteristics in regions of interest (ROIs) were examined with automated analysis procedures. Chronic hypoxia increased Ca2+ wave size in adults, rate of cytosolic Ca2+ removal in fetuses, and ROI recruitment in adults, albeit most ROIs occured in the same cell. Acute hypoxia decreased Ca2+ wave size in chronically hypoxic adults and decreased ROI recruitment in normoxic but not chronically hypoxic adults. It also reduced Ca2+ wave size in normoxic fetuses and obliterated Ca2+ signaling in chronically hypoxic fetuses. Maturation increased Ca2+ wave size as well as recruited more ROIs across the artery wall. These results indicate that chronic hypoxia modifies the influence of acute hypoxia on Ca2+ signaling in fetal and adult PAM in marked ways that may be important to pulmonary vascular dysfunction in those who live and are born at high altitude.

Support or Funding Information

Support from NIH, NSF, and the Walter E. Macpherson Society
AFP anti‐sense transcripts in mouse liver and their potential role in gene regulation
Authors: Dixon Maria S. , Qiu Guofang , Spear Brett T. , Peterson Martha L.
Published: June 2021

Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer, ranking the sixth most common cancer and third most common cause of cancer mortality worldwide. We have been studying mouse liver gene regulation to better understand mechanisms by which changes in gene expression contribute to liver development, homeostasis and disease. We identified Zinc Fingers and Homeoboxes 2 (Zhx2) as a regulator of alpha‐fetoprotein (AFP), a plasma protein that is highly expressed in the fetal liver but is shut off after birth. AFP expression is elevated in regenerating adult liver and hepatocellular carcinoma (HCC) and has been used extensively as a diagnostic marker of liver cancer. Interestingly, all genes that are misregulated in the absence of Zhx2 are also misregulated in HCC. Thus, to better understand gene regulation during HCC, we have been studying of the mechanism by which Zhx2 regulates gene expression. While studying AFP mRNA regulation by Zhx2, we identified previously unannotated antisense transcripts (asAFP) that partially overlap the 3′ half of the mouse AFP gene. asAFP RNAs are ~5kb alternatively spliced, mainly cytoplasmic, transcripts containing 2–4 exons that are likely to be non‐coding RNAs. These antisense transcripts were also detected in mouse liver RNA‐seq data. The abundance of asAFP RNA inversely correlates with AFP mRNA levels during postnatal development. Normally, asAFP RNA levels are high and AFP mRNA levels are low in the adult mouse liver. However, in the absence of Zhx2, AFP mRNA levels are higher and asAFP RNA levels are reduced. When portions of the asAFP RNA are over‐expressed in a liver cell line, endogenous AFP mRNA was reduced, suggesting asAFP transcripts repress AFP mRNA expression in trans. We will also knock‐down asAFP RNA and predict that AFP mRNA levels will increase. My central hypothesis is that asAFP RNA contributes to the post‐transcriptional regulation of AFP mRNA through an RNA‐RNA interaction. We are designing experiments to test this hypothesis and to determine at what level the regulation may occur.

Support or Funding Information

MCB‐1158234 from the National Science Foundation, R01‐DK59866 from the National Institutes of Health
Zhx2 and the Regulation of AFP Anti‐sense RNAs in Mouse Liver
Authors: Qiu Guofang , Dixon Maria , Spear Brett , Peterson Martha
Published: June 2021

Although the genomes of eukaryotes are mostly transcribed, only a very small percentage of the transcripts code for proteins. The majority of the transcripts do not have known functions. In recent years, growing evidence has suggested that these non‐coding transcripts may play important roles in gene regulation. In the past two years, our lab has identified and characterized long non‐coding RNAs that are transcribed from the anti‐sense strand of, and partially overlap with, the alpha‐fetoprotein (AFP) gene. AFP is highly expressed in fetal liver, but its expression decreases to a very low level after birth. In adult liver, AFP expression increases during regeneration due to liver damage or during the onset of hepatocellular carcinoma. Our study has shown that anti‐sense AFP (asAFP) transcripts are alternatively spliced and their abundance is inversely correlated with AFP mRNA during liver development. While these asAFP RNAs had not been annotated, RNA‐seq data from mouse liver confirmed and extended our characterization and ENCODE data for histone modifications and DNase I hypersensitivity show signals near the asAFP exons we have identified. Expression levels of AFP mRNA and asAFP RNA also are inversely correlated in the presence or absence of the Zinc fingers and homeoboxes 2 (Zhx2) gene. Zhx2, a putative transcription factor, was shown previously to contribute to the developmental repression of AFP; in the absence of Zhx2, AFP mRNA is high while asAFP RNA is low, compared to wild‐type. Transiently transfecting asAFP into a liver cell line causes a decrease in endogenous AFP, showing asAFP contributes to AFP regulation in trans. We have shown that the asAFP transcripts localize mainly in the poly(A)+ fraction of cytoplasmic RNA, but this does not identify where the asAFP RNA functions. To determine whether the asAFP RNA represses AFP mRNA in the nucleus or cytoplasm, we have been analyzing the abundance of these two RNAs in nuclear and cytoplasmic RNA from liver of Zhx2 wild‐type or knock‐out animals. To better understand the role of Zhx2 in modulating AFP and asAFP RNA expression, future studies will use chromatin IP to determine whether Zhx2 interacts directly with these genes. To identify which of the genes that are mis‐regulated in the absence of Zhx2 are direct targets, we plan to do Zhx2 ChIP‐seq experiments.

Support or Funding Information

MCB‐1158234 from the National Science Foundation, R01‐DK59866 from the National Institute of Health
Temporal Shifts in Differential Changes and Regulatory Relations among Multi‐Omic Assays in Social Defeat Mouse Model of Post‐Traumatic Stress Disorder
Authors: Muhie Seid , Gautam Aarti , Chakraborty Nabarun , Hammamieh Rasha , Meyerhoff James , Jett Marti
Published: June 2021

We collected hemi‐brain, hippocampus, amygdala, medial prefrontal cortex and ventral striatum from mice exposed to modified social‐stress for 1 to 12 days. We performed transcriptome, DNA‐methylome, miRNAome, and genome‐wide Creb1 transcription binding assays. Temporal and integrative analyses of the multi‐omics datasets were carried out using time‐dependent features extraction, and combinatorial algorithms. Our analyses showed unexpected shifts in relational time coordinates among the different omics datasets. Specifically, for a given gene or probe with significantly changed multi‐omics data points, we observed a temporal shift in its expression changes, methylation of its promoter, modulation of its transcript by upstream miRNAs, and binding of its promoter by transcription regulator. For many significantly changed features, the methylation changes occurred, unexpectedly, at earlier stress days while gene expression changes occurred at later time points. Many of the miRNA changes occurred at eight and later stress days. Our findings showed importance of temporal assays to capture time‐dependent multi‐omics changes as reliable signatures for a given condition. Integrative analyses of multi‐omics measurements based on a single time point may be deficient in many aspects, and may also be misleading if missed observations are interpreted as absence of changes.

Disclaimers: Research was conducted in compliance with the Animal Welfare Act, and all other Federal requirements. The views expressed are those of the author(s) and do not constitute endorsement by the U.S. Army.

Support or Funding Information

Funding: USAMRMC grant number 09284002
Simultaneously Revealing All Lysine Acylations in an Organism to Elucidate Substrate Metabolism
Authors: Nguyen Hong Hanh T. , Loo Joseph A. , Ogorzalek Loo Rachel R.
Published: June 2021

Within the past 8 years, lysine acetylation knowledge has extended greatly beyond one modification important for chromatin maintenance and transcription regulation to the whole family of reversible conserved short‐chain acylations emerging as an important regulatory mechanism coordinating metabolism in response to changes in nutrient availability. Inspired by these lysine acylation discoveries, we devise a mass spectrometry (MS) method to simultaneously reveal and evaluate all lysine acylations in an organism. Hypothesizing that an organism's pool of acyl‐CoA esters could suggest potential acyl‐lysine modifications hidden within MS/MS datasets, we chose two syntrophic bacteria, Syntrophomonas wolfei and Syntrophus aciditrophicus, for the study. They activate fatty acid and/or aromatic substrates with coenzyme A and stepwise convert acyl‐CoAs down to acetyl‐CoA for ATP production, thus containing a variety of acyl‐CoA intermediates. Different from the current approach of immuno‐enriching acylated peptide prior to MS, our shotgun non‐enrichment MS approach allows a quick, non immuno‐bias, economical and quantitative one‐step assessment of all lysine acylations within an organism, and the modification stoichiometries and their relative levels. We also showed characteristic ions are reliable to detect and confirm lysine acylations, establishing seven lysine acylations first seen in any organism and doubling the number of known short‐chain acylations. Our lysine acylation studies suggest a metabolism regulation mechanism in these syntrophs living at the thermodynamic limit of feasibility. This proteomic approach can be applied to any organism having a complex profile of acyl‐CoAs, especially anaerobic bacteria degrading fatty acids and aromatic compounds and gut bacteria producing butyrate.

Support or Funding Information

This work is funded by the US Department of Energy (UCLA Institute of Genomics and Proteomics, DE‐FC03‐02ER63421 to J. Loo) and the National Institutes of Health (R01GM085402 to J. Loo and R. Ogorzalek Loo).
Kinetics and Binding Studies Reveal that His‐40 is Essential for Catalysis in F420‐Dependent Glucose‐6‐Phosphate Dehydrogenase from Mycobacterium tuberculosis
Authors: Oyugi Mercy , Mayfield Joshua , Bashiri Ghader , Baker Edward , Zhang Yan , Johnson‐Winters Kayunta
Published: June 2021

Tuberculosis disease (TB) is a deadly infectious disease that is caused by Mycobacterium tuberculosis (Mtb). Our study focuses on F420‐dependent glucose‐6‐phosphate dehydrogenase (FGD), the enzyme that catalyzes the first committed step of the pentose phosphate pathway within Mtb. FGD uses the oxidized F420 cofactor to convert glucose‐6‐phosphate (G6P) to 6‐phosphogluconolactone, yielding reduced cofactor F420H2. F420H2 has been linked to the activation of a promising TB pro‐drug (PA‐824) and it also protects Mtb from nitrosative damages caused by the host's macrophage system. The FGD crystal structure revealed an active site His‐40 residue that has been proposed to be the active site base for the FGD reaction. We aim to probe the functionality of this His‐40 residue using site‐directed mutagenesis, crystallography, binding assays, circular dichroism, and steady state kinetic methods. Here, we present the results of the characterization of wild‐type FGD (wtFGD) and one FGD His‐40 mutant (FGD H40A). The circular dichroism data showed similar spectra for the FGD H40A and wtFGD, suggesting that the H40A mutation did not cause any structural changes within the enzyme. The steady state kinetics yielded kcat values of 1.40 ± 0.03 s−1 for wtFGD and 0.0016 ± 0.00 s−1 for FGD H40A, thus resulting in over 1000‐fold decrease in catalytic efficiency for the FGD H40A mutant. From the binding assays, we were able to conclude that His‐40 is not involved in G6P binding, but it helps in anchoring the F420 cofactor into the active site. The drastic loss in catalytic efficiency for the FGD H40A mutant led to crystallography experiments aimed at solving an FGD crystal structure with both F420 and G6P in the active site. FGD H40A crystals grew in 0.1 M ammonium citrate dibasic (pH 5.0) with 19% PEG 3350 as the precipitant, and diffracted to 2.9 Å to reveal the basis of substrate binding. In conclusion, the results presented here support the previously proposed mechanism which suggested that His‐40 is the active site base for the FGD reaction.

Support or Funding Information

This research was supported by NSF Grants 1120837
Metabolite‐Driven Modifications: Protein Acylations Elucidate Substrate Metabolism in Syntrophic Bacteria
Authors: Nguyen Hong Hanh T. , Nguyen Phuong H.N. , Gunsalus Robert P. , McInerney Michael J. , Loo Joseph A. , Ogorzalek Loo Rachel R.
Published: June 2021

Lysine acylation is a rising post‐translational modification (PTM) family that includes acetylation and similar short‐chain acylations. Reversible acylation has emerged as an important regulatory mechanism coordinating metabolism in response to changes in nutrient availability and metabolic fluxes. Since the levels of lysine acylations correlate with the levels of corresponding acyl‐CoAs and syntrophic bacteria produce complex central metabolic intermediates during fatty acid and aromatic compounds degradation, we used mass spectrometry to simultaneously reveal and evaluate all the acyl‐modifications in the syntrophs Syntrophomonas wolfei and Syntrophus aciditrophicus. Detected modifications provided insights into substrate metabolism regulations in these organisms living at the thermodynamic limits of microbial growth.

Support or Funding Information

This work is funded by the US Department of Energy (UCLA Institute of Genomics and Proteomics, DE‐FC03‐02ER63421 to J. Loo) and the National Institutes of Health (R01GM085402 to J. Loo and R. Ogorzalek Loo).
The Cornell NutriPhone: Enabling Precision Nutrition
Authors: Mehta Saurabh , Erickson David C.
Published: June 2021

The progress in modern technology and advances in biomedical engineering over the last few years have now made it possible to address a long‐term challenge of accurately and inexpensively assessing nutritional status at the point‐of‐care in both developed and developing country settings. This will facilitate improvements in designing and implementing nutritional interventions, generation of accurate representations of micronutrient status globally, and development and delivery of targeted nutritional programming.

The Cornell NutriPhone has resulted through a collaboration between scientists and engineers in Nutritional Sciences and Engineering at Cornell University in the time span of two years. When we began this partnership, our initial targets were to develop diagnostic tools that were affordable and accessible for populations in both resource‐limited and resource‐rich settings. From a technical standpoint, we also decided to focus on adapting conventional laboratory assays and gold standard methods rather than focusing on proxy biomarkers that are widely used in most field‐based rapid diagnostic tests.

We now have prototypes for low‐cost accurate quantitative determination of several micronutrients including vitamin D concentrations from a drop of blood at the point‐of‐care. Some of the tests are based on competitive antibody recognition, similar to other standard immunoassays. Instead of using an enzymatic or radioactive label however, the antibodies are conjugated to 30nm gold nano particles. The nano particles have a strong red color characteristic of their surface plasmon resonance. For the quantification of micronutrient levels, the brightness change is captured using the smartphone's camera after inserting the test strip in the smartphone accessory. The smartphone app then uses an algorithm to: scan for a region of most uniform color development, compute an averaged brightness difference, and estimate the concentration from the built‐in calibration curve.

We are currently conducting field validation studies in the U.S., India, and Ecuador to establish accuracy and reliability of the test and refine usability and interface of the software. In the short‐term, we are working on expanding the capability of the NutriPhone to other micronutrients and mutiplexing several nutrients on one chip/strip. Our long‐term vision is to equip health care facilities or community workers even in remote regions of the world with one such device to enable precision nutrition globally.

Support or Funding Information

National Science Foundation; Atkinson Center for a Sustainable Future
Attracting Attention: Parent Perceptions of Coverlines on HomeStyles Guides
Authors: Martin‐Biggers Jennifer T , Delaney Colleen , Povis‐Alleman Gayle , Hongu Nobuko , Worobey John , Byrd‐Bredbenner Carol
Published: June 2021

This study examined parent perceptions of coverlines used on 12 colorful 4‐page mini‐magazine‐style informational guides designed to improve weight‐related home environment and lifestyle practices in families with preschool children. Coverlines are short phrases designed to attract attention and motivate individuals to read articles in magazines. Health communication researchers developed coverlines based on results from content analysis of popular women and parenting magazine coverlines, asked 77 mothers of young children to rate how much each coverline would motivate them to read a short article related to the coverline, then selected the most highly rated coverlines for placement on guide covers. Examples of some highly rated coverlines are: “Breakfast is for champions! Feed your little super heroes”, “Secrets for successful family meals”, “Surprising links between family playtime and health”, “Who knew? Fruits and veggies are the new power foods”, “Make mealtime memories your kids will treasure”, “Smart, no‐cost family playtime ideas”, “Eat, play, sleep and love your family like never before”, and “Brilliant ideas for meals without squeals”. Coverlines were inserted in colorful text blocks positioned on the full‐color photograph forming the cover of the informational guides. Cognitive interviews were conducted with 117 English‐ and 112 Spanish‐speaking parents to determine their feelings about the coverlines and whether they believed the coverlines would make them and parents like them want to read the guide. The cognitive interviews of the coverlines were conducted after parents had read a guide. Almost all parents felt the coverlines would make them and other parents want to read the information in the guides. Parents also described the coverlines as “attention‐grabbing” and “catchy”. Participants made a few minor semantic change suggestions. Using motivating coverlines has the potential to increase intention and desire to read health promotion materials. Health communicators should consider incorporating evidence‐based, consumer‐tested coverlines in written materials to pique consumer interest.

Support or Funding Information

USDA NIFA #2011‐68001‐30170
Validity of anthropometric measurements, BMI and child growth assessments conducted by Head Start teachers: training needs for longitudinal child growth surveillance
Authors: Esquivel Monica Kazlausky , Fialksowski Marie K , Nigg Claudio R , Braun Kathryn L , Li Fenfang , Wilkens Lynne , Novotny Rachel Kazlausky
Published: June 2021

Background

High prevalence of childhood obesity and associated diseases have been observed in the US Affiliated Pacific but no system for monitoring young child obesity in the region is available. Head Start (HS) preschools located throughout the region complete bi‐annual growth assessments to assess childhood obesity and can potentially contribute to a growth monitoring system.

Objective

To assess the relative validity of HS anthropometric measurements and growth assessments for potential use in a surveillance system.

Methods

This was a validation study of HS child anthropometric measurements taken by HS teachers against standardized researcher measurements for height and weight. Data from 19 HS classrooms (135 children) that were previously enrolled in a larger randomized control trial were included in this study. Differences in height, weight, BMI and CDC BMI categories between HS teachers and a standardized researcher were analyzed using paired t‐tests, controlling for child age at time of measurement to quantify differences in HS and researcher measured heights and weights. weighted Kappa statistics determined agreement between the methods on BMI weight categories adjusting for age at measurement with post‐hoc paired t‐tests to analyze differences in BMI categories by measurement method.

Results

HS teacher growth assessments did not significantly differ from those of a standardized researcher and weighted Kappa statistics for agreement based on CDC BMI categories showed reasonable agreement between the two methods (weighted kappa=0.72).

Conclusions

At the programmatic level, this level of agreement suggests that HS measurements are adequate for determining prevalence of BMI categories. However, they may not provide sufficient precision to assess changes over time in individuals and research studies (ie: measuring intervention or program effect). Anthropometric data from HS centers may contribute to a system for childhood overweight and obesity surveillance in the US Affiliated Pacific, however, further training on conducting measurements would be necessary to track individual changes over time.

Support or Funding Information

This project is supported by the Agriculture and Food Research Initiative Grant no. 2011‐68001‐30335 from the USDA National Institute of Food and Agricultural Science Enhancement Coordinated Agricultural Program
Effect of obesity on liver metabolic and morphological profile in Zucker rats
Authors: Hakkak Reza , Korourian Soheila , Evans Teresa , Pavliv Oleksanda , Harb Alex , Melnyk Stepan
Published: June 2021

Fatty liver disease, ranging from simple hepatic steatosis and nonalcoholic staetohepatitis to cirrhosis, created a major health problem in the USA and world. Animal and human study showed that obesity can play important role in promotion of fatty liver. Our previous results demonstrated that obesity in Zuckar rats caused a significant morphological changes in liver: increase in fatty steatosis to small foci of mononuclear infiltration with perturbation in one carbon group metabolism donor S‐adenosylhomocysteine (SAM) and its product S‐adenosylhomocysteine (SAH) and SAM/SAH “methylation ratio”. The main objective of this study was to further investigate and compare metabolic profile of liver steatoss of lean and obese Zucker rats. After one week of acclimation, six‐weeks old female lean (n=26) and obese (n=20) Zucker rats were fed AIN‐93 G diet and sacrificed 155 days later. Liver samples were analyzed for free reduced glutathione (GSH), oxidized glutathione (GSSG), GSH/GSSG ratio, nitroglutathione (GSNO), and 3‐nitroTyrosine (3NT) using LC‐MS and HPLC‐ECD techniques. Our results shows that obesity caused: 1) a significant increase (p<0.05) liver steatosis; 2) a significant (P<0.05) decrease level of GSH and, on contrary, significant (P<0.005) increase level of GSSG; 3) crossing in glutathione level, that leaded to significant (P<0.0005) decrease of GSH/GSSG ratio, and 4) in contrast, significant (P<0.05) increase both of 3NT and GSNO. In summary, liver morphological and “methylation ratio” changes in obese rats are accompanied by significant oxidative and nitrosative stress. These metabolic changes must be consider in prevent of liver damages.

Support or Funding Information

(Supported by USDA and ABI grants to RH and SM).
Decreased beige adipocyte number and mitochondrial respiration coincide with reduced FGF21 gene expression in Sprague Dawley rats fed prenatal low protein and postnatal high fat diets
Authors: Claycombe Kate J , Vomhof‐DeKrey Emilie E , Garcia Rolando , Johnson William T , Uthus Eric , Roemmich James N
Published: June 2021

We have shown that protein malnutrition during fetal growth followed bypostnatal high‐fat diets results in a rapid increase in subcutaneous adiposetissue mass in the offspring contributing to development of obesity and insulin resistance. Recent studies have shown that the absence of a key transcription factor PR domain containing 16 (PRDM16) and fibroblast growth factor 21 (FGF21)are involved in conversion of precursor white adipocytes into more mitochondrial enriched and metabolically active beige adipocytes cause marked enlargement of the subcutaneous adipose tissue. Our hypothesis is that amaternal LP and postnatal HF diets increase the risk of development of obesity and insulin resistance in offspring, in part, by reducing the conversion of precursor white adipocytes into beige adipocytes in the subcutaneous adiposetissue of offspring. Using obese‐prone Sprague‐Dawley rats fed 8% low protein(LP) or 20% normal protein (NP) diets for 3 wk prior to conception and throughout pregnancy and lactation followed by 12 wks of 10% normal fat (NF) or 45% high fat (HF) diet feeding, we investigated whether prenatal LP and postnatal HF diets affect beige adipocyte number and oxidative respiratory function in subcutaneous adipose tissue. Results showed that subcutaneous adipose and liver fibroblast growth factor 21 (FGF21), PRDM16, and beigeadipocyte marker CD137, mRNA increase with postnatal HF diet in maternal NP group mice. In contrast, mice fed maternal LP and postnatal HF diets showed no increase in subcutaneous adipose tissue mitochondrial copy number, oxygen consumption rate, FGF21, PRDM16, and CD137 mRNA. These findings suggest that high‐fat diet fed offspring from mothers that consumed a low‐protein diet have reduced induction of beige adipocytes in subcutaneous adipose tissue and that this may be part of the mechanism by which maternal protein malnutrition causes offspring obesity and metabolic alterations.

Support or Funding Information

This work was supported by USDA Agricultural Research Service Project #3062‐51000‐052‐00D.
The Effect of Carotenoids on Transforming Growth Factor β1‐induced Fibrogenesis in LX‐2 cells, a Human Hepatic Stellate Cell Line
Authors: Bae Minkyung , Lee Ji‐Young
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the United States. In response to oxidative stress or inflammation, simple steatosis can progress to non‐alcoholic steatohepatitis (NASH), an advanced form of NAFLD. Fibrosis is one of key features of NASH with increased accumulation of extracellular matrix (ECM) in the liver. Hepatic stellate cells (HSCs) play important roles in the development of liver fibrosis as they are the primary source of ECM in the damaged liver. We previously demonstrated that astaxanthin (ASTX), a xanthophyll carotenoid, prevents the activation of HSCs and attenuated the expression of fibrogenic markers, such as α‐smooth muscle actin (α‐SMA) and procollagen type I α1 (COL1A1) in LX‐2 (a human HSC cell line) as well as primary mouse and human HSCs. The objective of this study was to compare the efficacy of other carotenoids, including lycopene, lutein, zeaxanthin and canthaxanthin, in inhibiting HSC activation with that of ASTX in LX‐2 cells. Lycopene is a carotene while the other carotenoids are xanthophylls with very similar structure. LX‐2 cells were treated with 0, 10, 25, 50 and 100 μM of each carotenoid for 24 h to measure cytotoxicity. While most cells were viable with all concentrations of ASTX and zeaxanthin, cells treated with 100 μM of lycopene, lutein or canthaxanthin were ~60–70% viable. To evaluate the effect of the carotenoids on the activation of LX‐2 cells by transforming growth factor β1 (TGFβ1), a potent fibrogenic cytokine, the cells were pretreated with 25 μM each carotenoid for 24 h, followed by the treatment with 4 ng/mL for 24 h. TGFβ1 treatment markedly increased mRNA of α‐SMA and COL1A1 in LX‐2 cells by ~4‐fold and 5‐fold, respectively. ASTX repressed TGFβ1‐induced expression of α‐SMA and COL1A1 by ~60% and ~30%, respectively. Lutein also decreased the expression of α‐SMA and COL1A1 by 85% and 70%, respectively. Basal and TGFβ1‐induced α‐SMA and COL1A1 protein levels were noticeably decreased by lutein and ASTX with lutein being more potent. TGFβ1‐induced COL1A1 protein expression was attenuated by lycopene, zeaxanthin and canthaxanthin, albeit less than lutein and ASTX. In conclusion, the results suggest that ASTX and lutein inhibit TGFβ1‐induced HSC activation and fibrogenic gene expression while zeaxanthin and canthaxanthin did not exert the same effect despite their similar structure to ASTX and lutein.

Support or Funding Information

United States Department of Agriculture, AFRI (2012‐67018‐19290)
Influence of temperature and humidity on the stability of carotenoids in biofortified maize genotypes
Authors: Ortiz Darwin , Rocheford Torbert , Ferruzzi Mario
Published: June 2021

Maize is a staple crop that has been the subject of biofortification efforts to increase the natural content of provitamin A carotenoids. While significant progress towards increasing provitamin A carotenoid content in maize varieties has been made, post‐harvest handling factors that influence carotenoids stability during storage has not been fully established. The objectives of this study were to determine carotenoid profiles of six selected provitamin A biofortified maize genotypes at various developmental stages, and assess the stability of carotenoids in maize kernels during controlled storage conditions (12 month‐period) including elevated temperature and relative humidity (RH). There were no significant changes in the content of individual carotenoids within genotypes during kernel development from 45 days after pollination through the time of harvest. Carotenoid losses through traditional grain drying were also minimal (<9%). However, stability of carotenoids in maize kernels over storage time after harvest was found to be dependent on both temperature and humidity, with variation observed among genotypes. Different forms of provitamin A carotenoids follow similar degradation rate. Carotenoid degradation rate appears to be independent of initial carotenoid concentration. Genotype (C17/DE3) had a degradation rate 2× faster than the other genotypes evaluated (P<0.001). These differences in carotenoid stability to controlled storage were attributed, in part, to observed differences in physical properties of the kernels (surface area and porosity). These results support the notion that effective control of moisture content and temperature of the kernels during storage conditions is essential to reduce the speed of degradative reactions.

Support or Funding Information

Funding provided by NSF, Harvest Plus, Colciencias and Fulbright‐Colombia.
Alpha‐Melanocyte Stimulating Hormone (α‐MSH) Affects Adipose Tissue Physiology in Broiler Chicks
Authors: Shipp Steven Lee , Wang Guoqing , Fu Xiangping , Cline Mark , Gilbert Elizabeth
Published: June 2021

The melanocortin system plays an important role in whole‐body energy homeostasis. In mammals, α‐melanocyte stimulating hormone (α‐MSH) induces satiety via the hypothalamus and stimulates lipolysis in adipose tissue. In chickens, it is also highly anorexigenic, but effects on adipose physiology have not been reported in an avian species. The objective of this study was to determine the effects of exogenous α‐MSH on adipose tissue physiology in broiler chicks. We hypothesized that peripheral α‐MSH injection would be associated with changes in transcriptional regulation of the melanocortin system, and lipid metabolism‐associated factors in adipose tissue. At four days post‐hatch, using a randomized complete block design with body weight as the blocking factor, individually caged Cobb‐500 broilers were assigned to receive intraperitoneal injection of 0 (vehicle), 5, 10, or 50 micrograms of α‐MSH (n=10/treatment) and euthanized at 60 minutes post‐injection and feed intake recorded. Subcutaneous and abdominal adipose tissue and Pectoralis major were collected for quantifying mRNA abundance of adipose lipid metabolism‐associated factors, and melanocortin receptors 1–5. Additionally, an in vitro study was conducted where the stromal‐vascular fraction of cells was isolated from the abdominal fat of 14 day‐old Cobb‐500 broilers. At 80% confluence, cells were treated with 0 (control), 1, 10, or 100 nM α‐MSH (n=3 independent experiments) and total RNA isolated at 4 hours to measure mRNA abundance of the same genes. Data were analyzed by ANOVA using the Fit Model platform of JMP (SAS Institute) and the statistical model included the main effect of treatment (sex was non‐significant for all variables and removed from the model), with Dunnett's test used for pairwise comparisons. Food intake was decreased (P < 0.0001) at 60 minutes in chicks that received 50 μg of α‐MSH as compared to vehicle. MC5R mRNA was increased (P < 0.05) in subcutaneous adipose tissue of chicks that received 10 and 50 μg of α‐MSH, but was not affected in the abdominal adipose or Pectoralis major, compared to vehicle‐injected chicks, and increased in vitro in cells receiving 10 and 100 nM α‐MSH, compared to control‐treated cells. The mRNA abundance of fatty acid binding protein 4 (FABP4) was also greater (P = 0.02) in 100 nM α‐MSH‐ than control‐treated cells. In conclusion, α‐MSH injection reduced feed intake, and was associated with increased gene expression of MC5R. Thus, in birds, peripheral α‐MSH may reduce feed intake, and effects in adipose tissue may be depot‐dependent and mediated through melanocortin receptor subtype 5.

Support or Funding Information

This research was supported by a National Science Foundation EAGER grant (IOS 1420285). Funding for this work was also provided in part, by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture.
Probing the Cell Cycle Significance of Cdt1 Phosphorylation at Novel Sites
Authors: Harris Regine Simone , Varma Dileep , Cook Jeanette Gowen
Published: June 2021

The cell cycle is regulated by proteins as it goes through the phases of G1, S, G2, and mitosis. DNA replication should occur once and only once in S phase. When re‐replication occurs erroneously it can lead to genomic instability such as cancer. Cdt1 is a protein that is highly regulated in the cell cycle and Cdt1 is degraded during S phase so that DNA cannot be re‐replicated. It plays a role in recruiting DNA replication MCM helicases to the origin recognition complex (ORC) in the G1 phase of the cell cycle. Cyclin dependent kinases (CDK) are regulatory proteins that phosphorylate Cdt1 for degradation by proteasomes in the S phase. Previous studies have investigated stress activated MAP kinase phosphorylation sites within Cdt1. Findings demonstrate that when those sites are mutated, such that they can no longer be phosphorylated, changes can occur in stability and function. Cdt1 is phosphorylated at two additional sites, Serine 93 and Threonine 152, but the functions of these phosphorylations are unknown. Our goal is to mutate those two sites to alanine, which cannot be phosphorylated, express the altered Cdt1 genes in human cells, and test for changes in Cdt1 behavior in a cell cycle dependent manner. We successfully created the S93A, T152A and S93AT152A (double mutant) Cdt1 expression plasmids by first, using PCR to create our inserts, then combining each mutant with a pENTR vector by Gibson assembly. The mutant Cdt1 constructs were transferred from the pENTR plasmids to pBabe‐puro plasmids by LR reaction for gene expression. For our transient transfection we used 293T cells to express our gene. Our stable Hela cell line was produced by retroviral transfection of 293T cells, then extracting the virus for transduction of Hela cells. Western blot analysis of 293T cells transiently transfected with these plasmids did not yield conclusive results with regard to the precise nature of the expression of the Cdt1 mutants. However, we are in the process of creating stable Hela cell lines using these plasmids. Our T152A and S93AT152A Hela cell line did not become stable as opposed to our S93A Hela cell line. We therefore suspect that our T152A mutant might have a negative effect on cell viability.

Support or Funding Information

NIH/NIGMS Grant #R25GM076162NSF Grant #DBI‐1038160
Water balance in desert birds ‐ challenges in a rapidly warming environment
Authors: Wolf Blair O.
Published: June 2021

Osmoregulation in desert birds involves a complex interplay of physiology, morphology and behavior that has been carefully documented over the last five decades. Rapid warming of the Earth's surface is producing significant new physiological challenges for many species; the potential capacity for individuals and species cope with these new challenges is of significant conservation interest. Here, we examine avenues and mechanisms of water intake and loss, and how warmer environments will affect the ability of birds in deserts to effectively cope with these challenges. Increased water requirements associated with greater demands for evaporative cooling may have differential impacts across species because of differences in body size, diet, and the evaporative mechanism employed. We show that adjustments in behavior and physiology, e. g. renal, performance may fail to compensate for these increased demands and result in changes in specie's distributions and community structure.

Support or Funding Information

Funding provided by NSF IOS‐122228
Potential adverse effects of polyunsaturated fatty acids: Influence of lipid oxidation on lymphatic transport of lipophilic bioactive components and cell morphology
Authors: Yao Mingfei , Kitamura Fuki , McClements David Julian , Xiao Hang
Published: June 2021

One of the most effective ways to enhance the bioavailability of highly lipophilic food components is to facilitate their chylomicron‐based lymphatic transport by oral administration with dietary lipids. However, lipids that contain polyunsaturated fatty acids (PUFA) are highly susceptible to oxidation. Ingestion of oxidized lipids is associated with numerous kinds of health risk, including diabetes, tumor development, and atherosclerosis. In addition, the oxidation of lipids may affect the absorption of other lipophilic bioactive components in foods. In this study, we compared the effect of oxidized and unoxidized linoleic acid (LA) on the transport of a highly lipophilic bioactive compound (5‐hydroxy‐6,7,8,4′tetramethoxylflavone or 5‐DMT) using a Caco‐2 cell model. Unoxidized LA improved the bioavailability of 5‐DMT by stimulating chylomicron production. Oxidized LA also enhanced the transport of 5‐DMT, however, it significantly affected the morphology of the Caco‐2 monolayer, and especially the tight junctions. Our results suggest that oxidized PUFAs may alter the transport pathway of lipophilic bioactives, which may influence their subsequent distribution and metabolism in the human body.

Support or Funding Information

USDA
Effects of a Dietary Strawberry Powder on Parameters of Vascular Health in Adolescent Males
Authors: Holt Roberta R , Djurica Dragana , Ren Jingyan , Hackman Robert M , Keen Carl L
Published: June 2021

Previous studies have shown that the intake of freeze‐dried strawberry powder (FDSP) improves select markers of cardiovascular health in at risk individuals; whether these improvements can be observed in adolescents is unknown. A randomized, double‐blind, crossover study enrolled 25 overweight or obese males, ages 14–18 years, to consume 50g of FDSP or an isocaloric control powder, daily for 1 week. Before and after each test period, measures of microvascular function, plasma nitrate/nitrite, and platelet reactivity were collected at baseline and 1 hour after FDSP intake. After 1 week of FDSP intake, an acute increase in the Framingham reactive hyperemia index (fRHI) was observed (p < 0.03). Plasma nitrate/nitrite levels significantly increased 1 hour after consuming the FDSP on the first study day prior to daily FDSP intake (p < 0.001) and during the second visit after one week of FDSP intake (p < 0.001). This increase in plasma nitrate levels was not observed with control powder intake. Taken together, the data from the present study supports the concept that strawberries can provide vascular health benefits to overweight and obese adolescent males.

Support or Funding Information

Research funding was provided in part by the California Strawberry Commission; USDA NIFA National Needs Graduate Fellowship (Djurica); and Agriculture and Food Research Initiative Competitive Grant (Holt).
Progression of antepartum depression differs by level of perceived social support and food insecurity
Authors: Natamba Barnabas Kahiira , Mehta Saurabh , Vermeylen Francoise , Stoltzfus Rebecca Joyce , Young Sera L , Griffiths Jeffrey K
Published: June 2021

Background

Antepartum depression (AD) is a risk factor for adverse pregnancy outcomes and may be worsened by food insecurity (FI) and lack of social support (SS). We studied the progression of AD from mid gestation to term and examined whether such progression differed by measures of SS and longitudinal measures of FI.

Methods

403 women were recruited (mean (±SD) gestational age =19.4 (±−3.8) weeks) and followed to term or dropout. At baseline, SS was assessed using a modified Duke‐UNC functional scale. Women were binned into 2 SS groups, based on scoring < or ≥ than the median SS value. FI (IFIAS) and AD (CES‐D≥17) were assessed at each visit using validated scales. Other variables were assessed at baseline using structured tools. Separately by SS category, we used mixed effects logistic regression models to examine the progression of AD; and, assessed whether such AD progression differed by longitudinal measures of FI.

Results

At baseline, 55.8% of the pregnant women belonged to the higher SS group. At this time point, the prevalence of AD among women in the low SS group (80.9%) was 2 times higher than similar levels in the higher SS group (40.4%). AD in the low SS group tended to decline with increasing gestational age but was always more prevalent than AD in the high SS group. In both SS groups, the progression of AD was positively associated with longitudinal measures of FI; and, the effect of FI on the progression of AD attained statistical significance among women in the low SS group. In the adjusted analyses, the probability of AD was three times higher among food insecure women in the low SS group than it was among food secure women in the high SS category.

Conclusions

We observed high levels of antepartum depression among women in northern Uganda that differed by social support and food insecurity suggesting the need for integrating mental health services that include food security and social support components into antenatal programs in this or similar contexts.

Support or Funding Information

This research was funded in part by Cornell University Weill Medical College Intercampus Seed Funds, K01 MH098902 from NIMH and USAID Cooperative Agreement AIDOAAL1000006
What Concerns and Questions Do Current Middle‐School Children Have When It Comes to Nutrition
Authors: Green Erin , Gaines Alisha , Hill Tisa Fontaine , Dollahite Jamie
Published: June 2021

This formative research explored the nutrition‐related questions and concerns of current middle‐school children. To date, few studies have explored nutrition and health perspectives of middle‐school aged children, an important group given that many food‐related behaviors and attitudes are being formed during this critical life stage. In‐depth semi‐structured interviews were conducted with 27 sixth to eighth grade students (12 male; 15 female) in New York State (14 rural; 13 urban). Interviews were audio recorded, transcribed verbatim, and analyzed using conventional qualitative content analysis to examine: 1) participants’ questions regarding nutrition, 2) current sources of nutrition information, and 3) nutrition and related health concerns. Results indicated that over half the sample reported having no current questions related to food or drinks. The most frequently reported sources of nutrition information included parents and school instructors or coursework, and a few children also cited coaches or grandparents as important sources. While some children did report accessing nutrition information online, none could recall specific websites frequented, with the exception of internet search engines (i.e. Google). Children reported looking up information related to the specific contents and characteristics of foods. Common themes around nutrition and related health concerns included weight status. Dietary restrictions like food allergies and concerns about what others such as siblings or peers were eating were also reported by some children. When discussing their concerns, children often provided a rationale that was not scientifically sound, indicating the need for additional nutrition information or education. These findings suggest middle‐school children without stated nutrition‐related questions often still have important concerns that may not be adequately addressed with current nutrition information seeking behaviors. This represents a potential opportunity to 1) design and disseminate nutrition‐related information targeted to both middle‐school children and their primary information sources (i.e. parents and schools), and 2) increase awareness of appropriate platforms providing nutrition‐related information.

Support or Funding Information

Funded by Agriculture and Food Research Initiative Grant no. 2012‐68001‐19604, USDA National Institute of Food and Agriculture, Childhood Obesity Prevention: Integrated Research, Education, and Extension to Prevent Childhood Obesity – A2101